17th International Photodynamic Association World Congress

We will discuss product target profiles that photosensitizers for PDT should met to satisfy the requirements of clinical PDT. Next, we will review the spectroscopy and photochemistry of the main families of photosensitizers with an impact in clinical PDT. We will critically examine the factors that made such photosensitizers appealing for PDT and their shortcomings. Finally, an overview of opportunities ahead will be presented. Topics addressed – The phototherapeutic window – Electronic spectroscopy of porphyrin derivatives, phthalocyanines and BODIPYs – Photochemistry and ROS – Factors contributing to the performance of PDT sensitizers

The two topics to be discussed related to the photon-tissue interaction including absorption and light scattering, correct definitions of energy or power density as well as the ability to convert these into a photon density. PDT efficacy determining parameters will be discussed including approaches to quantify them. Leading to the current status in monitoring PDT dose delivery. Topics addressed: – Photon - Tissue interaction – Known unknowns in PDT dosimetry – Intrinsic versus Explicit Dosimetry – PDT treatment planning: Where are we.

Topics addressed: – Global health problem-antibiotic resistance – General aspects of antimicrobial PDT – Appropriate photosensitizers – Applications of antimicrobial PDT

This presentation will discuss biological factors that act to determine the response of solid tumors to photodynamic therapy. It considers the biological consequences of prescribed illumination conditions, for example fluence rate effects on PDT-created cellular and vascular damage. An overview of PDT-initiated anti-tumor immunity will be provided. Finally, the role of the tumor stroma/microenvironment will be discussed in the context of its effect on PDT outcomes. Topics addressed: – PDT illumination and tumor cellular/vascular responses – Immunological aspects of PDT outcomes – Tumor stroma/microenvironment as determinants of PDT response

We will review approved photosensitizers in various jurisdictions including the indication for which they are approved and possible of label use of these photosensitizers. We review previous clinical trials which failed, focusing in particular on what lead to failures in some clinical trial. We discuss the need to have a physicist involved in the treatment planning and possibly even his presence during PDT delivery. Topics addressed: – Approved Photosensitizers and current clinical trials. – Failures of previous clinical trials – A physicist in the OR

Contamination of food takes place in all stages of production process, packaging, storage, transportation, sales and cooking. Large amounts of clean water are used throughout the food production process such as cleaning, sanitizing, peeling, cooling or cooking.In recent years Enterohaemorrhagic Escherichia coli (EHEC) caused a serious outbreak of food borne illness worldwide, where patients suffered from severe diarrhoea and in some cases from haemolytic-uremic syndrome. The source of infection for humans was contaminated food (e.g. bean sprout) with EHEC. Furthermore pigs, colonised with methicillin resistant Staphylococcus aureus (MRSA), has become an emerging risk to be a source of infection for humans. One new technology for successful eradication of bacteria is the photodynamic process. This talk summarizes the potential of photodynamic decontamination of foodstuff and decolonization of porcine skin as a novel approach for improving hygiene standards in the future.

Food safety challenges are present in modern life in many ways. The whole supply chain from harvesting, processing, storage and reaching consumers are full of contaminations possibilities. The problem causes a significant food los and waste. It is estimated today that about 30 % of the produced food never gets to the people. Besides that fact, we have today many examples of fresh food contamination like meet, chicken and fish, putting in risk the consumers. To control, in part, the situation, the abusive use of toxic substances, antibiotics and excess of acid water has been used causing a server for health and for environment. In this presentation we shall discuss the challenges ahead in this area and how photo reaction and photodynamic reactions are able to help to minimize the problems. Examples of project for fresh vegetables decontamination as well as for meat will be described in this presentation. The discussion about modification of food tast, composition or other characteristics will be presented as well as a study of the dynamic involving microorganism inactivation. Work supported by FAPESP, CNPq and CAPES – all Brazilian agencies.

Health experts estimate that every year food-borne illnesses in USA cost 86 billion dollars. Obviously, existing antibacterial technologies for microbial control of foods are not enough effective. Photosensitization is a treatment involving the interaction of the two non-toxic factors, photosensitizer and light, which in the presence of oxygen results in the destruction of the target microorganism. According to our results, chlorophyllin (food additive E140) exhibits perfect photosensitizing properties. After excitation with light (405 nm) it inactivates food pathogens, their spores, biofilms and yeasts/microfungi. This treatment significantly (2-3 log CFU/g) reduces microbial load on fruits, vegetables and sprouts without thermal effects and extended the shelf-life of treated produce by 2-4 days without reduction of nutritional value.

Photosensitization as a novel treatment has been effective against a wide range of food related microorganisms. Photoactivated curcumin, a plant based natural dye from the spice turmeric, has shown potential in extending the shelf life of fresh date fruit, oysters, salads and other minimally processed foods. It has been very effective against pathogenic bacteria such as E. coli 0157:H7, Listeria monocytogenes and Vibiro parahaemolyticus. It has significantly reduced the Aspergillus flavus conidia, hyphae and carcinogenic fungal toxin aflatoxin B1 in maize kernels. Photosensitization could be an efficacious and cost-effective method to inactivate a broad range of food borne pathogens.

This study evaluated phytotherapeutic compounds of Curcuma longa, Citrus lemon, Hamamelis virginiana and Hypericum perforatum, available on the market, as new photosensitizers (FS) in Dental Antimicrobial Photodynamic Therapy (aPDT). Each FS were analyzed in a spectrophotometer to determine the ideal light source. The ideal concentration of use, cytotoxicity on fibroblast culture, indirect reactive oxygen species (ROS) production was verified. Also, bacterial reduction was tested in culture of E. faecalis in planctonic form and in biofilm. Extracts of Curcuma longa, Citrus lemon, Hamamelis virginiana and Hypericum perforatum, showed potential for use in aPDT as photosensitizing agents.

We have treated more than 900 cases of CIN and uterine cervical cancer using PDT with Photofrin(P-PDT). PDT is considered to be a treatment modality with higher fertility sparing capacity than cone resection as uterine preservation therapy for CIN3. P-PDT has high efficacy for CIN and has low obstetric risks such as premature birth after PDT. However, the side effect of photosensitivity is strong, so it has not reached standard treatment. Therefore, in order to investigate the safety and effectiveness of L-PDT using Laserphyrin with less half-life in blood, we started Phase I/II study of L-PDT. After approval of the ethical committee, phase I/II study of L-PDT was performed on 43 cases of CIN2-3 with IC. Although temporary lower abdominal pain and fever were observed in L-PDT, photosensitivity was hardly seen unlike P-PDT, suggesting safety. CR rates 3 and 6 months after L-PDT were 95 and 98%, suggesting efficacy. These data suggested L-PDT would become the next generation PDT for CIN.

Photodynamic therapy (PDT) is one of cancer treatments using a specific photosensitizer and a particular type of light. Immunotoxin (IT), on the other hand, has been developed as a molecular targeted therapy as a drug of malignant tumor. However, the internalization of IT into cytoplasm remains as the big issue. We developed a new technology which target the cancer specific membrane protein by IT in combination with PDT. We present here the use of a saporin-conjugated antibody against the tumor specific membrane protein, Robo1, and a photosensitizer. Though the treatment with IT only showed little effect on cancer cells, the addition of light illumination augmented the cytotoxic effect several tens of times. By longer exposure, we observed a significant effect even on a cancer cell line which has ten times less Robo1 expression. These results suggested that the combination of IT and PDT widen the therapeutic measure both in the specific drug delivery and in the drug target.

Background: Patients with centrally located early lung cancer (CLELC) are often heavy smokers with a high risk of multiple lung cancer (MPLC); treatment strategies must preserve the cardiopulmonary function. Methods: Between June 2013 and June 2018, 10 patients with CLELC underwent photodynamic therapy (PDT) at Nippon Medical School Hospital. Results: Eight patients underwent surgery for primary lung cancer and then underwent PDT for the treatment of secondary primary CLELC, two patients were performed PDT alone. In 6 of these 10 patients, CLELC was found as metachronous lesions and in 4 patients as synchronous lesions using sputum cytology and a bronchoscopical examination. Among the 10 patients with MPLC including lung cancers which were resected by surgery, all 10 CLELC lesions exhibited a complete response after PDT. Conclusions: PDT can play an important role for the treatment strategy for MPLC.

Multidrug resistance is a process in which cancer cells utilize ATP-binding cassette (ABC) transmembrane transporter to actively pump anti-cancer agents out of the cells. In a panel of human breast cancer cell lines, we showed multiple ABC transporters play a role in the transport of benzoporphyrin derivative (BPD) photosensitizer and confer resistance to BPD-based photodynamic therapy. To overcome these challenges, we introduced porphyrin-lipid nanovesiclesas a new strategy to escape ABC transporter-mediated efflux of BPD for improved PDT outcomes.

As salvage surgery after failure of chemoradiotherapy (CRT) for esophageal cancer shows high morbidity and mortality, curative and less invasive salvage treatment has been needed. Photodynamic therapy (PDT) is a candidate for local failure after CRT. Taraporfin sodium (Leserphyrin) is the new generation photosensitizer which was made in Japan, and has an advantage of low skin toxicity because of rapid clearance. We conducted multi-institutional trial to evaluate the safety and efficacy of PDT using taraporfin sodium for patients with histologically proven local failure limited within the muscularis propria after 50Gy or more CRT for esophageal cancer. The PDT procedure commenced with intravenous administration of a 40mg/m2 dose of talaporfin sodium followed by diode laser irradiation at a 664nm wavelength. We set the primary endpoint as local complete response (L-CR) per patients. In this study, 26 patients were enrolled and all were treated with PDT. Twenty three patients were assessed L-CR after PDT; the L-CR rate per patient was 88.5 % (95% CI: 69.8%－97.6%). No skin phototoxicity was observed only with two weeks’ sun shade, and no grade 3 or worse non-hematological toxicities related to PDT were observed. At the timing of all enrolled cases were followed 3 years or longer, the median local progression free and overall survival time were 3.1 and 4.2 years, respectively. PDT using new generation photosensitizer and a diode laser is a safe and potentially curative salvage treatment for local failure after CRT for patients with esophageal cancer.

Optical agents with long-lasting luminescence after removal of light excitation are promising for ultrasensitive in vivo imaging due to minimized tissue background. However, such imaging agents are rare and generally limited to inorganic nanoparticles. In this study, we introduce a new generation of purely organic agents based on semiconducting polymer nanoparticles (SPNs) that can store photon energy via chemical defects and gradually emit near-infrared (NIR) afterglow luminescence at 780 nm after light irradiation. We demonstrate the unique abilities of afterglow SPNs for background-free luminescence cancer imaging, evaluation of drug-induced liver toxicity, imaging-guided therapy, and ultrasensitive in vitro diagnostics.

Two Functional luminescence nanoparticles will be introduced. The first type of materials is upconversion nanoparticles (UCNPs). I will present new developments regarding engineering UCNPs towards optogenetic applications in cancer immunotherapy and visual enhancement. The second is a type of FRET-like organic Biodpy nanoparticles that were tailored with outstanding NIR absorbing ability to enhance PDT effect. Rather than the conventional laser light needed in PDT, I will present their ultralow power lamp operable PDT applications in deep tissue tumor treatment.

Biodegradable polymeric nanoparticles (Bp NPs) show low toxicity, good biocompatibility, therefore making it a good candidate for the delivery of photosensitizers to increase the bioavailability and improve therapeutic outcomes of PDT. Two types of Bp NPs, Polylactic-co-glycolic acid (PLGA) nanoparticles (NPs) and chitosan / methoxy polyethylene glycol - polylactic acid (CPP) nanoparticles were prepared to load the photosensitizer 5- aminolevulinic acid (ALA) and zinc pthalocyanine (Zp), respectively. The characteristics, safety and effectiveness were determined in vitro and vivo. Bp Nps were spherical with smooth surfaces and high encapsulation efficiency. Bp NPs increased the production of fluorescence and reactive oxygen species by photosensitizers and enhanced the effectiveness of PDT for SCC. No obvious toxicity was observed in vivo. Bp NPs provide a promising photosensitizer delivery strategy for the topical photodynamic therapy of cutaneous SCC.

Brain cancer is causing an alarming level of morbidity and mortality in the world. Photodynamic Therapy (PDT) has recently gain attention in Gliomas treatment. The implementation of PDT for brain tumors, and especially glioblastoma (GBM), has already been approved in some countries. Conjugated polymer nanoparticles (CPNs) have massive potential in PDT area. Recently, our group developed metallated porphyrin-doped CPNs for PDT of Gliomas tumor cells trough ROS damage generation. In a next step, the study of a "delivery system" that allows the systemic circulation of CPNs, no recognition of the mononuclear phagocyte system and that in turn provides specific uptake in tumor cells within the CNS would be promising and with great clinical potential. Trojan horse therapy using CPNs-loaded monocytes to improve tumor penetration of nanoparticles was investigated and successfully achieved.

Biocompatible silver sulphide quantum dot-cetuximab nanoconjugates (QD-cetuximab) were investigated as a potential contrast agent for pancreatic cancer biophotonic diagnostics. Using fixed immunofluorescence staining, QD-cetuximab targeted the EGFR-positive pancreatic cancer cell lines PANC-1 and CFPAC-1 at concentrations up to 600µg ml-1. Using live immunofluorescence staining, similar targeting efficacy was observed at concentrations up to 50µg ml-1, but a reduction in targeting was seen at higher concentrations. Thus, QD-cetuximab nanoconjugates have the potential to target live EGFR-positive pancreatic cancer cells at doses up to 50µg ml-1. The reduction in fluorescence observed at higher concentrations is likely to be secondary to cetuximab toxicity.

Vascular anomalies are mainly divided into two types: vascular tumors and vascular malformations. The clinical manifestations of vascular anomalies which involving kind of multiple systems are quite complicated. Patients with vascular diseases generally visit different departments which are sometimes improper. In order to diagnose and treat vascular anomalies better, relevant disciplines should strengthen communication. Therefore, it is necessary to establish multi-disciplinary team of vascular anomalies, including department of dermatology, plastic surgery, invasive technology, ophthalmology, radiology, ultrasound, pathology. Our MDT of vascular anomalies already consulted 20 patients and the patients acquired the right and effective treatment.

Cancer patients often must confront the decision of whether to continue high dose chemotherapy at the expense of cumulative toxicities and high cost. Here, we introduce a dual pronged approach to modulate the microenvironment of desmoplastic pancreatic tumors and enable significant dose de-escalation of the FDA approved frontline chemotherapeutic nanoliposomal irinotecan (nal-IRI) without compromising long-term tumor control. We demonstrate that light-based photodynamic priming (PDP) coupled with vitamin D receptor (VDR) activation targets cancer-associated microvasculature and fibroblasts to increase intratumoral nal-IRI and suppress pro-tumorigenic CXCL12/CXCR7 crosstalk, resulting in improved tolerability with retained efficacy. Strategies aimed at modifying the tumor landscape to increase susceptibility remains a promising and relatively underexplored approach and may simultaneously improve patient outcomes and quality-of-life.

Glioblastoma is a malignant brain tumor with a poor prognosis. Currently, complete resection is rarely feasible, since tumor cells usually infiltrate the surrounding brain. The INDYGO clinical trial has been achieved to assess the toxicity of intraoperative photodynamic therapy to treat newly diagnosed Glioblastoma. Today, we believe that the PDT effect can be improved by a higher light dose. The DOSINDYGO clinical trial aims to achieve a light-dose escalation, increasing up to four times the initial light dose used in the INDYGO trial. These light doses should allow to treat deeper in tissues and thus decrease the recurrence risk.

The use of photodynamic therapy (PDT) as a novel neo-adjuvant treatment alone and in combination with the cytotoxic agent 5- azadeoxycytidine (5ADC) against breast cancer cells was investigated. In vitro PDT treatment resulted in early cytotoxicity while 5ADC treatment elicited delayed cytotoxic effects with synergy in combination. In vivo studies using an orthotopic 4T1 breast cancer mouse model demonstrated local and distant antitumour effects of liposomal verteporfin-PDT in comparison to 5ADC alone and in combination with this agent. Flow cytometry and gene expression analysis provided evidence for PDT-mediated activation of innate immunity.

Vascular targeted photodynamic therapy (V-PDT) has been widely used for the treatment of port wine stains (PWS) in China with great success,but the efficacy of V-PDT for PWS are still varied in individuals. In this prospective study, the vascular features of PWS patients were evaluated by optical coherence tomography angiography (OCTA). The results showed that OCTA is a promising noninvasive method to evaluate the depth, and vascular density and the vessel diameter of PWS lesions before and after V-PDT, which may help to optimize the V-PDT treatment parameters.

Background: Hematoporphyrin monomethyl ether photodynamic therapy (HMME-PDT) is considered a vascular-targeted therapy, which is a promising alternative treatment for PWS. Objective: We evaluated the efficacy, adverse effects, and redarkness of PWS using HMME-PDT. Methods: From March 2017 to January 2018, a total of 81 Chinese patients of PWS with negative HMME skin test results were recruited and each treated area were observed during 10 months of follow-up. Results: Among the investigated factors, we found that the clinical efficacy was related to the type, size, color and texture of the lesions, and the number of treatments. Meanwhile, the adverse effects were related to laser treatment used before PDT, and fewer treatments were associated with higher redarkness rate. Conclusion: Although Chinese patients have a good response to vascular-targeted HMME-PDT, more attention should be paid to the adverse effects and long-term follow-up.

Invasive grade IV malignant gliomas remain difficult to treat with a typical survival time post-diagnosis of around 15 months. The main challenge in treating glioblastoma lies in the killing of cancer cells that remain in the brain parenchyma after the resection of the main primary tumor as recurrences quickly develop around the resection cavity borders leading to patient death. Experimental and clinical studies demonstrate that PDT can complement the existing traditional tumor therapies consisting of surgical resection, radiation therapy and chemotherapy. We have developed a photosensitizer nanogel (NPM-GEL) for local delivery in the GBM resected cavity. NPM-GEL was made MR imageable by doping with MR contrast agent. The cytotoxicity of NPM-GEL following irradiation at 20J was studied on different GBM cell lines (T98-G, U-87 EGFRvIII, and C6). In vivo toxicity study showed NPM-GEL was well tolerated when injected in rat brain. Intracranial distribution was studied and optimized using

The purpose of our study was to increase the targeting efficiency of tetraphenylporphyrin (TPP) using silica (SiO2) nanoparticles (NPs) to tumor site through the enhanced permeability and retention (EPR) effect. In a nude mice model, SiO2-TPP NPs showed high anticancer efficacy against HT-29 cell subcutaneous tumor xenograft when irradiated compared to free-carrier TPP. Tumor volume and weight were analyzed along with H&E, Ki67, cleaved caspase-3, TUNEL staining and biodistribution. This improved in vivo antitumor efficacy seemed to be due to the targetability of NPs. These results demonstrate that our new SiO2-TPP NPs are useful for tumor-targeted cancer treatment by PDT.

Photodynamic therapy (PDT) is used to treat cancer. However, sometimes repeated PDT can lead to the development of treatment resistance. Cancer stem-cells (CSCs) are cells that give rise to new cancer cells. They have high expression of drug-transporters that help to remove cancer killing agents from the cells. As CSCs are known to drive resistance of other cancer treatments through these transporters, we hypothesize that these CSCs underline PDT resistance. We first generated PDT resistant cells and our results indicate that CSCs are enriched in PDT resistant cells. In future studies we will determine if CSCs contribute to PDT resistance.

OBJECTIVE In this study on the effectiveness and safety of photodynamic therapy (PDT) using talaporfin sodium and a semiconductor laser, 30 consecutive patients with newly diagnosed glioblastoma were analyzed and compared with those of 164 patients treated without PDT during the same period. RESULTS The median PFS times of the PDT and control groups were 19.6 and 9.0 months, with 6-month PFS rates of 86.3% and 64.9%, respectively (p = 0.016). The median OS times were 27.4 and 22.1 months, with 1-year OS rates of 95.7% and 72.5%, respectively (p = 0.0327). Multivariate analyses found PDT, preoperative Karnofsky Performance Scale score, and IDH mutation to be significant independent prognostic factors for both OS and PFS. CONCLUSIONS The results suggest that PDT with talaporfin sodium and a semiconductor laser provides excellent local control and potential survival benefits for patients with newly newly diagnosed glioblastoma

Theranostic tumor targeted nanoconstructs are, in theory, the ideal constructs for image guided surgery of glioblastoma (GBM) followed by in situ photodynamic combination treatments of unresectable disease. However, the complexity of their in vivo behavior and ambiguous specificity hampers their theranostic potential. By developing near-infrared (NIR) Paired Nanotracer Imaging (PNI), in conjunction with an intelligent EGFR-targeted NIR-active nanoconstruct, we quantified the in vivo specificity of the nanoconstructs for GBM tissue in vivo. As such, we present the first report of a complex GBM-targeted NIR-active nanoconstruct for photodynamic combination treatments with quantifiable molecular specificity in vivo, which simultaneously provides superior optical contrast for image-guided surgery.

In recent years, talaporfin sodium-mediated photodynamic therapy (NPe6-PDT) against glioblastoma (GBM) has been developed; however molecular mechanism of NPe6-PDT-dependent GBM cell death has not yet been well elucidated. Therefore, we investigated the molecular machinery of GBM cell death induced by NPe6-PDT using human GBM cell lines. As a result, activation of the stress signaling and suppression of the survival signaling were observed during NPe6-PDT-dependent GBM cell death. In addition, we succeeded in establishment of GBM cell lines with acquired NPe6-PDT resistance and currently investigating further about the machinery which regulates NPe6-PDT resistance in these cells.

The use of nanoplatforms to enhance photosensitizer buildup in tumor is a promising strategy to optimize PDT efficacy. We evaluated the viability of membrane fusogenic lipossomes as vehicles for protoporphyrin IX for in vitro non-melanoma skin cancer PDT. PpIX uptake was assessed and compared to the internalization of free PpIX in non-melanoma skin cancer cells and healthy fibroblasts. PDT efficacy and ROS generation was evaluated with MFL-PpIX and free PpIX for different incubation times, PpIX concentration and light fluences (630 nm) in both cell lines.

The authors demonstrated the efficacy and safety of additional intra-operative photodynamic therapy (PDT) using talaporfin sodium and a semiconductor laser for patients with malignant meningeal tumors. Eight patients (1 newly diagnosed and 7 recurrent) with grade II or III meningeal tumors were enrolled. Almost all patients demonstrated a long clinical history, undergoing multiple surgeries and radiotherapy prior to PDT. Except for 1 patient who had a short survival time, local control was achieved by resection of the tumor bulk and additional PDT without any adverse events. The authors thus demonstrated the clinical feasibility of PDT for malignant meningeal tumors.

This study aims to establish a stable and efficacious photoimmuno-nanoconstructs platform delivering the tunable lipid nanoconstructs in heterocellular 3D nodules containing MIA PaCa-2 PDAC cells and patient-derived pancreatic cancer-associated fibroblasts to demonstrate the specificity, penetration and targeted phototoxicity of the optimal targeted nanoconstructs. The designed NIR light activatable targeted photoimmuno-nanoconstructs (TPINs) demonstrate an effective binding to MIA PaCa-2 and PCAFs nodules with up to 8-fold specificity with respect to the nontargated nanoconstructs, higher penetration and up to 16-fold enhancements in the selective photodynamic destruction of cancer cells into heterocellular 3D model of MIA PaCa-2 and PCAFs.

The limited penetration of laser radiation into biological tissue prevents the widespread distribution of PD and PDT methods to clinical practice. We have investigated several approaches for PD and PDT of deep-seated tumors: - Diagnostics and navigation of tumors with fluorescence excitation in the red and near infrared spectral ranges; - Optical fiber ports and catheters. Clinical trials; - Photosensitizers and fluorescent dyes with fluorescence in far red and near infrared spectral range; - Joint action of radiopharmaceuticals and photosensitizers based on Cherenkov radiation; - Action through photodynamic inactivation of tumor-associated macrophages and microglia. Studies have been conducted on experimental animals with grafted tumors and, in part, on cancer patients in the clinic.

Photodynamic therapy (PDT) is an alternative in the treatment of cancer. This study was to evaluate the cellular viability and the intracellular localization of Fotoenticine (PS) in the treatment of gliosarcoma. Cytotoxicity tests, morphology, and PS localization were performed. It was observed 100% viability in the dark and 0% in the highest concentrations after PDT, the mitochondrial activity test showed a reduction after PDT. The EROS production was higher in the PDT groups and dependent on the PS concentration. Morphologically, after PDT, cytoplasmic destruction was observed, rounded cells without nuclear alterations. The PS accumulated in the lysosomes, mitochondria, and cytoskeleton.

ALA + MRgFUS has been shown, noninvasively, to significantly slow the growth of transplanted gliomas in rodent brains. We sought to optimize results in a C6 intracranial rat glioma tumor model. Seven days following tumor implantation, ALA was injected before MRgFUS which was delivered for 20 min to single or multiple points (16) (MPS). The tumor growth response for animals receiving ALA, FUS, ALA + FUS and a sham control group were evaluated weekly. Only the 5-ALA + FUS groups showed tumor response and extended survival. The MPS method resulted in complete cessation of tumor growth.

Mesoporous silica nanoparticles are potential drug delivery systems for biomedical applications due to their biocompatibility, tunable size, easy functionalization, and high chemical stability. In this work, they will be used as nanocarriers for organic photosensitizers in order to improve their solubility, delivery and targeting to be implemented in photodynamic therapy. The photosensitizers are grafted in the nanoparticles´shell for photodynamic therapy, together with short chains of polyethylene glycol to enhance their stability in water. As a result, well-dispersed silica nanoparticles with good singlet oxygen production are obtained. “In vitro” experiments are being carried out to check their phototherapy activity in HeLa cells.

Photodynamic Therapy (PDT) is used to enhance the extent of tumour resection, enhancing survival. However, high-grade tumour types are heterogenous and their susceptibility to PDT may vary greatly. This in vitro study looked to determine what if any dose-response variation exists in neuroblastoma, human keratinocyte and glioblastoma multiforme cells by comparing the degree of cell kill achieved using increasing 630nm laser light doses alongside different photosensitizers. These different tumours demonstrated significantly different behaviours with certain photosensitizers, while others produced a uniform response. This has implications for future clinical research and may suggest a mechanism beyond simple free radical species generation.

In this study, we have developed a new cranial window implantation technology using thin, flexible, and transparent, biocompatible film as a window material. These cranial windows can offer enough-wide area accessibility for light illumination for PDT and pre- or post-treatment longitudinal imaging monitoring over extended periods of time.

Chemotherapy is one of the most common treatments for hyperproliferative diseases, but its efficacy and tolerability are often limited by off-target toxicity. A promising approach to improve these therapies is to activate drugs exclusively at their desired place of action. Light is a powerful tool in this respect, as it has unparalleled properties as a regulatory signal for pharmacological applications. We report here on the development and characterization of phototrexate, the first light-regulated inhibitor of the human DHFR, as a photochromic analog of methotrexate. Our results show that phototrexate behaves as a potent antifolate in its photoactivated configuration, and that it is nearly inactive in its dark-relaxed form.

We investigated the usefulness of ICGm, a novel nano-drug delivery (nano-DDS) system composed of polymeric micelle and indocyanine green for near-infrared (NIR) fluorescence imaging and photothermal therapy (PTT) using an orthotopic murine model of neuroblastoma. ICGm was selectively accumulated in the lesions, and temperature-controlled PTT exerted antineoplastic effects without damage to the surrounding organs. Most of all the treated cases showed the therapeutic depth reached to 5 mm, and the maximum depth of around 10 mm was obtained. These results suggest that ICGm is a promising novel theranostic nano-DDS for neuroblastoma.

Striving to develop novel heavy-atom free photosensitizers for photodynamic therapy underpins the current work. BODIPY-anthracene/pyrene/perylene photosensitizers have been shown to be selectively activated in response to environmental polarity to generate singlet oxygen via triplet states formed from charge-separated excited states. Both the singlet oxygen generating capabilities and fluorogenic response can be modulated by the substitution pattern. Water-soluble derivatives were synthesized via fluorine substitution reactions and in vitro studies indicate cytotoxicity at micromolar concentrations in human breast cells, indicating the potential of these systems to be used at photodynamic agents.

Guaiazulene is emerging as a photosensitizer. We aimed to measure singlet oxygen in vitro using a guaiazulene+red laser. Guaiazulene and erythrosine were prepared as an experiment and positive control, respectively. Then, both groups were irradiated with a red laser. Singlet oxygen was measured by ESR and analyzed with significant differences at p-value < 0.05. The average singlet oxygen of 5 µM significantly higher than 15 µM group (p < 0.01). Both 4 and 8 J/cm2 resulted in relatively equal singlet oxygen amounts. Five µM of guaiazulene activated by 625 nm laser at 4 - 8 J/cm2 dramatically generated singlet oxygen.

Recently, handheld spectroscopic probes were introduced enabling quantitative analysis of 5-aminolevulinic acid (5-ALA) induced fluorescence intensity (FI). In the current ex-vivo study, we investigated the value of such probes in samples of diffusely infiltrating gliomas derived from 5-ALA fluorescence-guided surgery. According to our data, we found significantly different median FI values between different visible fluorescence levels, different WHO grades (WHO grade IV/III vs WHO grade II) and specific histopathological parameters of anaplasia. Consequently, handheld spectroscopic probes represent a powerful tool for visualization of intratumoral glioma heterogeneity and will thus optimize in future glioma surgery.

Aminolevulinic acid–based photodynamic therapy (ALA-PDT) has emerged as a new alternative for chemotherapy in cancer treatment due to its high specificity and low side effects. We added siRNAs and inhibitors of transporters to study the roles of transporters in ALA uptake in sets of normal and cancer cell lines. PEPT1 and PAT1 were expressed only in normal lung and prostate cells, respectively, but not in their cancerous counterparts. Inhibition of these transporters showed a significant decrease in PpIX production only in normal cells. PEPT1 and PAT1 transporter inhibitors could be possible new drugs to increase the specificity of ALA-PDT.

In this presentation we will report on some of the key optical properties of a novel water soluble chlorin photosensitizing agent (YLG-1), which is synthesized by Guilin Hui-ang Biopharmoceutic Co. Ltd. of China. We characterized the photosensitizer on the basis of absorption and fluorescence emission, and time resolved fluorescence in various solutions. The effects of photobleaching were probed to characterize its decay kinetics. Singlet oxygen quantum yield was also evaluated. Additionally, its antitumor and antimicrobial potentials were evaluated in in vitro and in vivo models. Preliminary results suggest that this novel chlorin-based photosensitizer has great potential in photodynamic therapy.

To quantify singlet oxygen formation among various irradiation modes. Ninety ul of 1, 10 and 100 µM Azulene (in DMSO) + 10 µl of 10 mM dimethyl anthracene was added to 96-black-well-plate. After 30 minutes, 638-nm-LED irradiations were performed (0.5 watts) to obtain 4 or 40 J/cm2 by either continuous, fractional or pulse mode. Singlet oxygen detection in a Varioscan fluorescence microplate reader (excitation/emission wavelength = 375/473 nm) was conducted and optical density was recorded. One-way-ANOVA+multiple comparison was performed at a statistical different level at p < 0.05. The continuous mode resulted in relatively higher singlet oxygen amount.

During glioma surgery the neurosurgeon faces the challenge of distinguishing between normal tissue, infiltration zone and the tumor core. Although 5-ALA induced Protoporphyrin IX (PpIX) fluorescence guided surgery has improved the delineation of the tumor border significantly, it is difficult to distinguish different intratumoral areas based on visible fluorescence alone. As recently described we propose lifetime imaging of PpIX for enhanced tumor visualization and tumor area discrimination. The lifetime is elevated in both tumor core and infiltration zone and can be clearly distinguished from normal tissue. Ultimately, this may support the neurosurgeon to achieve a maximal safe glioma resection.

Despite suggestions that 5-aminolevulinic acid (ALA) could be used for pancreatic cancer photodiagnostics, some pancreatic cancers do not exhibit ALA-induced fluorescence. The pancreatic cancer cell line PANC-1 exhibits poor ALA-induced fluorescence. We compared the mRNA expression of haem biosynthesis pathway proteins with or without 24 hours ALA incubation between pancreatic cancer cell lines PANC-1 and CFPAC-1 and control cell line H6c7. PANC-1 demonstrated minimal expression of ALA influx transporter PEPT1 and significant upregulation of PpIX efflux transporter ABCG2. These transporters are likely to be responsible for the weak fluorescence observed in PANC-1, which may have relevance to other ALA-resistant cancers.

Here we present a series of novel (aza)phthalocyanines with charged (cationic, anionic) as well as non-charged peripheral substituents. These compounds were directly compared to several clinically approved photosensitizers. Studied (Aza)Pcs were taken up to the cells by endocytosis and could be detected in endolysosomal compartment. Under the irradiation they displayed diverse photodynamic activities in vitro (EC50 values from 10-4 M to 10-9 M) causing damage that led to a predominantly necrosis-type cell death. Furthermore, (Aza)Pcs have shown consistently very low toxicity in the absence of light. Main causes of variable effectiveness of (Aza)Pcs in vitro were identified as 1) diverse cellular uptake and 2) interaction of some (Aza)Pcs with serum albumin. Water-solubility and high activity also indicate the possibility of application in vascular-targeted photodynamic therapy.

Delivering singlet oxygen to the site of interest is challenging because of its short lifetime. As an alternative to introducing sensitizers directly into tissue, we have developed a technique by which the sensitizer particles are sequestered in the plastron of a superhydrophobic surface, thereby preventing contact between sensitizer and tissue, allowing singlet oxygen to diffuse to biological surfaces. We describe two approaches to maximizing the concentration of singlet oxygen released from the superhydrophobic surface: sensitizer support morphology and device conformance. The results of size and porosity of the support particles, and 3D-printing to conform to biological structures will be presented.

Here we report a man with Epidermodysplasia verruciformis who had had wart-like lesions almost over his body for about 34 years and developed Aks and Bowen's disease on his head, lower arms, shoulder and neck. He had already had several SCCs excised and resulted in many scars on his head. To reduce the incidence of scar and squamous cell carcinoma, topical 5-aminolaevulinic acid photodynamic therapy was used. All the lesions responded to the treatment and showed a complete clinical clearing after 3-5 sessions of PDT. ALA-PDT is useful for the treatment of AKs and Bowen’s disease in Epidermodysplasia verruciformis.

In this work, we used the time-resolved, high sensibility and remote measurement characteristics of Thermal Lens Spectrometry to investigate the relation between the photoconversion rate of protohypericin to hypericin with initial concentration and light intensity. The results show a linear relation of the photoreaction rate with concentration of Protohypericin, indicating that the overall reaction includes steps comprising distinct intermediates formation. In addition, we present the TLS relevance to photochemical characterization of photosensitive drugs in low concentration range.

The efficient synthesis of a photostable low molecular weight carboxamide bacteriochlorin with sub-nanomolar EC50 in vitro after light exposure offers the possibility to treat skin lesions with topical administration. When topically applied to the skin of mice bearing CT26 subcutaneous tumors, followed 2h later by illumination at 750 nm, this photosensitizer significantly impacted the kinetics of tumor growth. Also, the pharmacokinetics shows a fast clearance from the body (t1/2 ~3h) diminishing the risk of off target reactions. Ongoing work aims at the topical treatment of melanoma tumor models.

Photodynamic Therapy is an alternative therapy for brain tumors, associating a photosensitizer, light, and molecular oxygen, generating reactive oxygen species. This work evaluates the cytotoxic action of PDT with Aminolevulinic Acid (ALA) and Fotoenticine, in gliosarcoma cells (9L/lacZ). The best incubation time was 4h, to ALA (125 ug/ml) AND 1h to Fotoenticine (200 ug/ml). After PDT, viability decrease in 95.2% with ALA treatment, and 94.6% with ftoenticine, when compared to control. ALA is widely used for the treatment of brain tumors, but the Fotoenticine can be an alternative, since with less time of incubation presented similar result to the ALA.

Cancer-associated fibroblasts (CAFs) are important components of the tumor microenvironment. CAFs affect the biological behavior of tumor cells and play critical roles in growth, invasion, and metastasis of tumor. Topical 5-aminolevulinic acid-mediated photodynamic therapy (ALA-PDT) is established approach for non-melonoma skin cancers and can induce effective antitumor immune response. While, the role of CAFs in ALA-PDT for cSCC is less known. This study investigated the effects of CAFs during ALA-PDT on cutaneous squamous cell carcinoma (cSCC).

Background. UVB is thought to be a major cause of DNA damage, which is the most important mechanism of UV-induced skin photodamage in epidermal cells. Although several studies found that ALA-PDT can prevent the occurrence of photodamaged dermatosis, the exact mechanism is still unknown. Aim. We conducted this experiment to verify the protective effect of ALA-PDT on UVB-induced photodamage and explore its mechanism. Methods. In vivo animal experiments, the dorsal skin of hairless mice were treated with ALA-PDT or saline, and then exposed to 180 mJ/m2 ultraviolet. For in vitro experiments, HaCaT cells were treated with ALA-PDT or untreated, and then exposed to 60 mJ/m2 UVB irradiation. Result. We found that ALA-PDT pretreatment can both reduce apoptosis, inhibit proliferation, accelerate the clearance of CPDs and activate p53 in vivo and vitro, which indicated that ALA-PDT pretreatment can induce a protective DNA damage response, thereby protecting against UVB-induced photodamage.

Thioxanthene dyes are interesting as potential sensitizers for photodynamic therapy, since they have a sufficiently high quantum yield of triplets (up to 0.9), absorb light in the visible region of the spectrum, have normal and thermally activated delayed fluorescence’s and phosphorescence, which can be used for diagnostics of the distribution of the dye in body tissues. Xanthene dyes, especially modified, are easily removed from the body. At the Institute of Technology (St. Petersburg, Russia), the xanthene dyes with a heavy atom in their structure, with internal triplet sensitizers to increase the quantum yield of triplet states, have been synthesized and investigated their photophysical properties. Also studied xanthene derivatives with annulated cycles for shift the absorption to the long wavelength region.

For the skin aging, we prepared cosmetic cream of four containing Ce6 conjugated photosensitizers. The cream has even dispersion and good stability in vitro. To check anti-wrinkle efficacy in vivo, we have included two group i.e. normal diet and high-fat diet mice. The prepared creams from these compounds were then applied to the BALB/c mice skin after removal of the hair and after the light was given to the applied area. Ce6 conjugation cream was the most effective. Our results showed that the cream may serve as an ideal anti-aging agent in photodynamic therapy.

Studies report that there is a strong relationship between temperature and porphyrin synthesis in biological tissue. In this study, menthol, methyl nicotinate and ginger extract (thermogenic and/or vasodilator substances) were incorporated into either the ALA or methyl-ALA cream to investigate the PpIX production in rat skin. The methyl nicotinate was the one with the highest optimization effect of PpIX production after three hours of incubation of the cream. These results are promising as a possible strategy for decreasing the incubation time of the precursor cream in various clinical protocols and increasing the photosensitizer production in lesions.

A novel ester between aminolaevulinic acid (ALA) and the iron chelating agent CP94 has been synthesised (AP2-18) and evaluated in a range of cultured human primary cells. Iron chelation achieved via CP94 or AP2-18 administration consistently increased PpIX accumulation but the benefits of enhancement on PpIX-PDT cell kill were most pronounced when lower doses of ALA or MAL were utilised. AP2-18 also significantly enhanced both PpIX accumulation and PDT cytotoxicity experimentally in both skin cancer and glioma cells. Newly synthesised AP2-18 is therefore concluded to be an efficacious prodrug for PpIX-induced PDT that warrants further investigation.

ICG lactosome, a drug delivery system (DDS) composed of polymeric micelle and indocyanine green (ICG), shows a selective accumulation in cancer tumor based on EPR effect. Since ICG is known to be a near-infrared absorbing substance, when a tumor in which ICG accumulate is irradiated with a near-infrared (NIR) light, only the tumor can be heated by -reaction. We confirmed that ICG lactosome was selectively accumulated in the tumors, and NIR irradiation efficiently raised tumor temperature. Furthermore, the tumors were eliminated with a probability of almost 100% when the peak of tumor temperature during NIR irradiation reached 43°C or higher.

5-aminolevulinic acid-based photodynamic therapy (ALA-PDT) is being widely used in cancer therapy owing to the tumor-specific accumulation of photosensitizing protoporphyrin IX (PpIX) after the administration of ALA. However, the mechanism of tumor-specific PpIX accumulation is not well understood. The purpose of the present study was to identify the mechanism of tumor-specific PpIX accumulation. We found that high expression of PEPT1 (ALA influx transporter) and low expression of ABCG2 (porphyrin efflux transporter) determined ALA-induced PpIX production and cellular photosensitivity in vitro. Moreover, the amount of mitochondrial iron ions were lower in tumor cells causing high PpIX accumulation.

Photothermal therapy (PTT) is an effective cancer treatment. We investigated the therapeutic effect of the temperature-controlled PTT system combined with a near infrared absorbing agent, IR783 and with using the 808 nm near-infrared laser on orthotopic hepatoma model rat. Laparoscopic imaging revealed the lesion of hepatoma. The visualized tumors was irradiated by the laser, being kept the tumor temperature of 45°C. PTT using IR783 induced cell death and inhibited the growth of hematoma. IR783 is useful as a photoabsorbing agent and temperature regulated PTT using IR783 is effective treatment for rat hepatoma.

Protein inactivation by reactive oxygen species (ROS) is considered to trigger cell death pathways associated with protein dysfunction; however, the detailed mechanisms and direct involvement in photodynamic therapy (PDT) have not been revealed. Thereby, we report Ir(III) complexes designed for ROS generation through a rational strategy to investigate protein modifications by ROS. The Ir(III) complexes were effective as PDT agents with low-energy irradiation because of the relatively high 1O2 quantum yield, even with two-photon activation. In addition, two types of protein modifications (protein oxidation and photo-crosslinking) involved in PDT were characterized by mass spectrometry. Consequently, we present a plausible PDT modality that utilizes photo-activation of rationally designed Ir(III) complexes, indicating the feasibility of a better optimized Ir(III) complex for PDT.

Summary: Complete safe tumor resection is the initial therapy of choice in spinal ependymomas and 5-ALA fluorescence is a promising technique to facilitate this goal. In this currently largest patient series we could demonstrate, that 5-ALA induced fluorescence was present in the majority of spinal ependymomas. Moreover, residual fluorescence could be detected in a significant number of cases even after assumed complete conventional ependymoma resection under white light and histopathological analysis of remaining fluorescent areas verified presence of residual tumor cells in all cases. Our data therefore suggest that application of 5-ALA may improve the outcome of spinal ependymoma surgery.

Photodynamic therapy (PDT) is a technique based on the photo-activation of a sensitizer to induce cell death. It is mostly recommended for superficial basal cell carcinoma with complete response rates between 50 to 80%. Moreover, this response rate drops significantly for thicker and nodular BCCs, mainly due to the limited light penetration into tumor. In this study, optical clearing agent was used to improving the light distribution and the PDT response in BCC lesions. OCT imaging showed that OCA doubled the depth of light penetration and no side effect was observed in the patients during and 30 days after PDT.

This multicenter study analyzes the impact of intraoperative 5-aminolevulinic acid (5-ALA) fluorescence for improved visualization of brain metastases (BM) to overcome incomplete resections. 5-ALA was administered in 157 brain metastases. The fluorescence quality and homogeneity were investigated and correlated with primary tumor and histopathological subtype. Visible fluorescence was observed in 104 BM (66%) with a frequently heterogeneous pattern (84%), whereas no fluorescence was detected in 53 cases (34%). Fluorescence was less visible in BM of melanomas (p=0.037) and more common for ductal breast cancer subtype (p=0.008). Thus, our data indicate that 5-ALA fluorescence is beneficial for intraoperative visualization of BM.

The Warburg effect is the phenomenon where tumors consume higher levels of glucose than normal cells. We analyzed the efficacy of PDT with glucose-conjugated chlorin e6 (G-Ce6), developed a new photosensitizer, for canine mammary gland tumors via in vitro and in vivo studies. In the in vitro study, cell viability was observed to decrease in a G-Ce6 concentration- and light intensity-dependent manner in the PDT group. In the in vivo study, PDT induced significant tumor regrowth delay. PDT using G-Ce6 may be beneficial in the treatment of dogs with mammary gland tumors.

Objective: To observe the efficacy of ALA -PDT in treating vulval CA in pregnancy. Methods: The efficacies of ALA-PDT on 16 cases of CA in pregnancy as well as cryotherapy on 22 cases of CA in pregnancy were analyzed. Results: The treatment group showed a wart clearance rate of 93.8% after 3 PDT treatments, while the control group 72.7% after 3 cryotherapy treatments. After the 3-month follow-up, the treatment group registered a recurrence rate of 6.3%, whereas the control group 36.4%, indicating a statistically significant difference . After the 1-month postpartum follow-up, the newborns grew and developed well, without any abnormality in physical examinations. Conclusion: ALA-PDT is safe and effective in treating CA in pregnancy.

ALA-PDT inducing pyroptosis of sebocytes via activation of NLRP3 inflammasome pathway Human SZ95 sebocytes were treated with ALA-PDT at different dosages. Real-time PCR was implemented to observe the expression of NLRP3 and Caspase-1 mRNA. ELISA was applied to detect cytokines release after ALA-PDT, and western blotting was used to analyze pyroptosis related target. It found that ALA-PDT on sebaceous glands in vitro triggered reactive oxygen species (ROS) generation leads to formation of NLRP3 inflammasome and Caspase-1 activation, resulting a rapid sterile inflammation and pyroptosis-dependent IL-1β secretion. Therefore, oral antioxidants taken before ALA-PDT may affect the treatment outcome.

We compared MB-PDT effect on two breast cancer cell lines: MDA-MB-231 from a triple negative adenocarcinoma, and T47D from a ductal cell tumor of woman. Methylene blue concentrations at 5, 10 and 20 μM and red light doses of 20, 40 and 60 J/cm2 were employed. Cell viability was evaluated with the MTT test, obtaining around 80% of inhibition with 20 μM and 60 J/cm2 of light dose.

In the present work, we show the in vitro effects of rose bengal (RB) as a photosensitizer (PS) and green light to eliminate two breast cancer cell lines: MDA MB231, a negative triple line highly aggressive and invasive, and T47D a luminal line of the infiltrating ductal type. RB concentrations of 0.5, 1 and 5 μM and radiation fluences of 2.5, 5 and 10 J/cm2 were evaluated. Cell viability was evaluated with the MTT test obtaining ~92% inhibition for MDA MB231 and ~82% for T47D with 5 μM and 10 J/cm2. Many of the cell treated showed apoptotic death signals.

In this project we demonstrate the ability of our new type of quantum dots (QDs), Vivodots™ nanoparticles, a safe and biocompatible type of QDs, to enhance the efficiency of 5-aminolevulinic acid (5ALA) when linked to the surface of the particle. We demonstrate this effect using cell cultures and in vivo models, and discuss the various underlying mechanisms. Our findings would offer a new basis for effective and broad utilization of 5ALA in image guided surgery, facile diagnostics and photodynamic therapy.

Persistent infection with Human papillomavirus (HPV) has been identified as the major cause of the Cervical Intraepithelial Neoplasia (CIN), a precursor of cervical cancer. This is a controlled non-randomized clinical trial for CIN 1, 2/3 treatment with photodynamic therapy. The total rate of complete response was 75% for CIN 1 (two years after PDT), 21.43% for placebo group, 67% and 90% for CIN 2 and 3, respectively (one year after PDT). The results of hybrid capture are showing a significant decrease (70-80%) in viral load. To improve the results for HSIL treatments, was coupled a laser fiber to illuminate the endocervix.

Aim of the study was to determine the immune effect of PDT with hypericin (HY-PDT) used in sublethal doses on the secretion of Interleukin 8 and 10 by experimental models of residual colon cancer cells in vitro. Results HY-PDT amplified the secretion of IL-8 by SW620 cell line, but the decrease of IL-8 secreted by the SW480 cell line. The increase in secretion of IL-10 was noticed in the SW480 cell line, but the changes of secretion IL-10 by SW 620 was not noted. SW480 cell line without PDT secreted higher levels of IL-8 and IL-10 than SW620 cells. Based on these research findings it could be told, that PDT both eliminates and control primary tumors using cytotoxic effect while HY-PDT at lower doses can modulate function of tumor microenvironment by releasing interleukins depended on metastatic activity of tumor cells.

The aim of this study was to determine the immune effect of PDT with 5-aminolevulinic acid (ALA-PDT) used in sublethal doses on the secretion of growth factors: VEGF, GM-CSF, G-CSF, FGF , Interleukin 6,8 and 10 and S100 protein secretion by experimental models of residual colon cancer cells in vitro. ALA-PDT amplified the secretion of GM-CSF and IL-8 by both cell lines. The decrease in secretion of VEGF, G-CSF IL-6, IL-10 and FGF was noticed in the SW620. SW620 line cells secreted higher levels of FGF and G-CSF, IL-6, IL-8 and IL-10, while SW480cells more actively released GM-CSF. After application of ALA PDT the S100 protein concentration was reduced in both cancer cell lines. No effect of ALA-PDT was noted on VEGF secretion by the non-metastatic SW480 cells.

Summary Vulvar Lichen Sclerosus (VLS) is a chronic, noninfectious, non-neoplastic dermatosis with probable autoimmune origin that affects the anogenital region. The main clinical symptom is vulvar pruritus and the recommended treatment is topical corticosteroid. Hydropic degeneration, hyperkeratosis and collagen hyalinization are epithelial morphological changes observed in VLS. PDT showed to be effective for clinical symptom control, in addition improved epithelial morphology, comparing to topical corticosteroid.

Mitochondria play a fundamental role in generating of energy in cells. Electron transfer in electron transport chain (ETC) involved in mitochondria was one of the biological functions of heme. Heme biosynthesizes increased upon the addition of 5-aminolevulinic acid (ALA). Complex I and II protein expression were downregulated, while complex IV and cytochrome c expression were upregulated by the addition of ALA. Administration of ALA significantly increased Complex IV (COX) activity and cellular ATP level. Surprisingly, the mitochondrial content and mitochondrial membrane potential were unchanged. Consistently, the relative mRNA-expression of transcription factors affecting mitochondrial biogenesis was unchanged after the addition of ALA.

To investigate the distribution of ALA in Cervical lesion tissue by tissue frozen section and fluorescence microscopy. 20% of 5-ALA were topically applied to the cervix.Biopsies were taken from the lesions for histology examination, Fluorescence microscopy was performed methods in these frozen section of cervical tissue to observe the distribution of ALA and the result was analyzed  semi-quantitatively. The maximal Protoporphyrin IX （PpIX）fluorescence was observed in the most serious tissue with cervical lesions .fluorescence photometry corroborated these results.

5-aminolevulinic acid-photodynamic therapy is known to be effective in several skin diseases .Mechanisms of ALA-PDT to treat these skin diseases still remain elusive. In this study, we aimed to investigate mechanism of ALA-PDT in in-vitro and in-vivo models. For in vitro, we use human keratinocyte cell line (HaCaT) cells. For in vivo, we use 20 rabbits to induce hyperkeratosis acne model in their ear. Dermatoscope was used to see follicle hyperkeratosis and skin biopsy to analyze histology and immunohistochemical of PKC, FGFR2b, K1, K6 and K16. Results from this study suggest that ROS stimulated by ALA-PDT lead to inhibition of FGFR2b pathway in PKC downstream to cause reduction of IL1α expression, and eventually, keratinocytes differentiation and proliferation. Our data thus reveal a treatment mechanism of ALA-PDT underlying hyperkeratosis related dermatoses.

Multidrug resistance (MDR) is a significant reason for the failure of chemotherapy, especially in ovarian cancer. The first players implicated in MDR are drug transporters from the ATP binding cassette (ABC) drug transporters family, which are ATP-driven transmembrane efflux pumps in cancer cells. A presumably promising strategy to overcome chemoresistance is targeted inhibition of ATP production in tumor cells. In this study, we developed a mitochondria-targeted photodynamic therapy (PDT) approach to overcome chemoresistance in ovarian tumors. The same PDT agent was also used to improve surgical outcome by intraoperative imaging.

Cancer therapy success depends on the selectivity towards cancer cells. Combinatorial therapies have been studied to treat cancer. Here we present a combination of photodynamic therapy with a glycolysis inhibitor. In cancer cells, glycolysis metabolism is altered with an increased glycolysis rate (Warburg effect), and several molecules have been studied to inhibit this pathway. 3-Bromopyruvate (3BP) inhibits hexokinase an enzyme of glycolysis pathway. In vitro combination studies of redaporfin-PDT and 3BP were performed in CT26 (colon carcinoma – mouse) and NIH/3T3 (fibroblasts – mouse) cell lines. A higher selectivity towards cancer cell line compared to non-cancerous was found. In vivo studies in mice are being performed.

In recent years, accompanying the progress of diagnostic imaging, opportunities for small pulmonary lesions to be detected have been increasing. We investigated whether marking of small pulmonary lesions that are difficult to identify in the lung periphery is possible using low power laser. The result is that the laser light could be clearly confirmed from the pleural surface. Pathologically, no damage was found in the lungs at the laser-irradiated site. Low power laser could be safely used intraoperatively to identify the site of small pulmonary lesions in the peripheral lung.

Indocyanine green (ICG) is a photothermal agent, photosensitizer which shows specific accumulation in hepatocellular carcinoma (HCC) cells. We recently developed a photodynamic therapy (PDT) using ICG and near-infrared (NIR) laser as a new anti-cancer treatment for HCC. HuH-7 cells, a well-differentiated human HCC cell line, were transplanted subcutaneously into mice xenografts model for in vivo experiment. ICG was administered 24h before NIR irradiation. The irradiation was performed at three tumor locations by 823-nm NIR laser on days 1 and 7. The mean tumor volume on day 9 was significantly smaller after ICG-NIR irradiation compared to tumor without irradiation (p=0.01).

Photodynamic therapy (PDT) is well known to be effective as a treatment for early-stage lung cancer located in central airway. But the lesion area is hard to observed clearly by conventional white light bronchoscopy (WLB) alone. Autofluorescence bronchoscopy (AFB, OLYMPUS CORPORATION) can be improved visualization of malignant lesions. It cannot be observed in magenta by AFB after Talaporfin Sodium (Leserfrin®, Meiji Seika Pharma Co., Ltd.) administration. ABF is not helpful for tumor localization diagnosis just before the PDT, and requires careful observation by WLB and photodynamic diagnosis (PDD) by another device.

Objective: This study aimed to study the correlation among oxidization, autophagy and apoptosis in photodynamic therapy. Methods: PDT induced oxidization, autophagy and apoptosis was systemically studied. And the interaction among the effects was studied by using inhibitors of autophagy (3-MA), apoptosis (ZVAD-fmk) and oxidization(NAC). Results and conclusion: PDT could induce autophagy in HUVEC. Blocking the oxidization pathway could affect cellular response (either autophagy or apoptosis) to PDT. Inhibition of authophagy could lead to increased PDT induced cell apoptosis. On the other hand, inhibition of apoptosis could enhence PDT induced cell autophagy.

Although multimodality treatments have been attempted, malignant pleural mesothelioma (MPM) still has a high mortality. It is valuable to detect a new treatment for MPM. We already experimented whether a combination treatment consisting of pemetrexed (PEM) chemotherapy and photodynamic therapy (PDT) using the photosensitizer NPe6, enhanced the antitumor effect for MPM. PEM treatment followed by NPe6-PDT resulted in a reduction in the tumor size and inhibited re-growth. Next, we plan to conduct a combination treatment of immune checkpoint inhibitor (ICI) and PDT. We expect the synergistic effect of the combination treatment of ICI and PDT.

In the present study we for the first time characterized the effects of low-dose PDT on MSCs, which represent key component of stroma and play a pivotal role in tumorigenesis. We found that PDT-mediated intracellular ROS induction significantly improved angiogenic potential of MSCs and modified MSC interactions with leukocytes. PDT-mediated cytoskeleton reorganization decreased MSC motility and chemotaxis. Low-dose PDT resulted in rapid activation of Erk1/2, JNK, Akt2 and inhibition of GSK-3β signaling in MSCs, which can represent a possible mechanism underlying effects of low-dose PDT on stromal cells. Thus, our study demonstrates that role of stromal cells shouldn’t be underestimated during development and optimization of PDT.

Chlorin e6 as a well-known photosensitizer was tested as its highly pure trisodium salt (Fotolon®) in a phase IIb clinical trial, in patients with recurrent NSCLC after first line treatments. In 16 patients the safety, tolerability and efficacy of the photosensitizer was monitored in a monocentric GCP-Trial. Additionally a pharmacokinetic assessment of the substance in blood plasma was performed. After only one single treatment the enrolled patients showed best response between day 28 and 85. Histologies from biopsies remained tumor free until day 85. The safety profile was excellent with no drug related serious events

Cancer cells reprogram their metabolism to promote tumorigenesis and increase drug resistance, through the Warburg Effect. The impact of PBM on the Warburg Effect and antineoplastic treatment such as PDT in glioblastoma cells was investigated. Dose dependent PBM mediated decreased glycolysis, increased oxidative phosphorylation, altered cellular proliferation rate, and induced a biphasic response in mRNA expression of multiple cancer genetic markers. These changes are reflected in the efficacy of antineoplastic therapies, here TLD1433-mediated PDT, whereby the efficacy changes also showed a biphasic response, which was not affected by CCO and ROS inhibition, suggesting a CCO and ROS independent mechanism.

Sequential damages in specific organelles are key to improve the efficiency of PDT protocols however the specific pathways involved in cell survival or death and/or adaptive responses to damages in specific organelles are unknown. By using a photosensitizer 1,9-dymethyl methylene blue (DMMB), which accumulates specifically and damages both mitochondria and lysosomes, we aimed to dissect the cellular responses linked to autophagy and other catabolic processes. To understand further the modulation on cell processes, we have performed gene expression, proteomics and lipidomics analysis to elucidate the molecules and pathways related to cell death signaling.

We investigated the molecular effects of PDT using inhibitors to impair DNA damage response (NCA) and redox (E3330) functions of APE1 protein. The hypothesis is that inhibition of APE1 would sensitize cancer cells to PDT. HeLa (cervix) and A549 (lung) cells were photosensitized by chloroaluminum-phthalocyanine into a nanoemulsion (with/without NCA-500 µM or E3330-50 µM) and irradiated (0.5 J/cm2) @670 nm. No alterations in cytotoxicity was observed adding the inhibitors to PDT when conducted in A549 cells. However, the redox inhibitor E3330 increased the effects of PDT in HeLa cells. These results stated that APE1 protein is critical to PDT response.

Photodynamic therapy (PDT) has been used for selective identification and treatment of various cancers, but it has never been explored as a combination approach for image-guided surgery and PDT for the treatment of prostate cancer. In this study using a PSMA-targeted PDT agent PSMA-1-Pc413, we found that this theranostic agent was able to provide guidance for surgery. More importantly, adjuvant PDT further eliminated tumor recurrence, leading to significantly extended survival. These results demonstrate a potentially significant improvement for conventional radical prostatectomy, suggesting that adjuvant PDT can effectively ablate invisible cancer cells following image guided surgery and therefore improve surgical outcome.

The high frequency of Hepatocellular Carcinoma (HCC) patients and the unsatisfactory results obtained with conventional treatments, encouraged investigations for alternative treatments such as photodynamic therapy (PDT). In this study, we evaluated the efficacy of 5-Aminolevulinic acid (5-ALA) mediated PDT on HCC and the influence of 5-ALA PDT on human immune system. Our preliminary results demonstrated that PDT can be effective against HCC cell lines, inducing a dose-dependent cell death in a pattern related to the p53 expression profile. Interestingly, conditioned media of PDT-treated HCC cell lines induce a decrease of HCC cell proliferation and an increase of human PBMC proliferation.

Photodynamic therapy is proposed for glioblastoma mainly for its capacity to kill tumor cells and to potentially induce an immune activation. Our study aimed to evaluate the impact of serum exosomes produced before and after ALA-PpIX-PDT on immune cells. Immunodeficient rats were xenografted with U87 cells and treated with multifractionated light. MRI revealed an elevated diffusion values in the center of the PDT-treated tumor. High exosome levels were detected in rat’s serum after xenograft while a strong decrease were noticed after PDT treatment. Pre-treated exosomes had an immunosuppressive effect on PBMC, while post-treated exosomes induced a sustained immune cell proliferation.

Optimizing the parameters of PDT for specific cancer cells by identifying the mechanisms involved in the process represents a considerable tool for the development of photosensitizer delivery systems and the improvement of PDT targets for cancer. Proteoglycans, important constituents of extracellular matrix (ECM) play an important role in both progression and inhibition of tumor development. This study determines by gene expression whether proteoglycan are associated with PDT resistance observed in human epidermoid larynx carcinoma cells (HEp-2). We conclude that PDT increases the expression of DECORIN and BIGLYCAN genes, involved in metastatic dissemination, leading to inhibitory effect growth in Hep-2 cells

The photodynamic activity (PDT) of Tetra triethyleneoxysulfonyl zinc phthalocyanine (ZnPc) was studied on HepG2 and Huh-7 cells a typical example of hepatocellular carcinoma (HCC). HCC cells showed a dose dependent uptake of ZnPc. Photo-activation of ZnPc (0.5-5 µM) in HCC cells revealed strong PDT effects, leading to a dose-dependent decrease of cancer cells without any sign of re-proliferation and a dose-dependent increase in caspase-3-activity. By contrast, non-photoactivated ZnPc did not induce any cytotoxicity. The formation of ROS and free radical detected in the cytoplasm/nucleus of HCC cells and the expression of apoptotic proteins confirm the apoptotic mode of cell death.

We recently reported that an effective interstitial photodynamic therapy (I-PDT) treatment is associated with concurrent light-induced tissue heating (LITH). We investigated the interaction of photoreaction and levels of heat observed during I-PDT, by conducting in-vitro studies where heat and photoreaction were independently regulated. We utilized the crosslinked signal transducer and activator of transcription-3 to evaluate the photoreaction dose during heat, for the first time. This knowledge will allow advancement of the utilization of I-PDT in the treatment of locally advanced cancer.

Background: In 1967 a few years after the first working laser was invented, Endre Mester in Semmelweis University Budapest, took some mice, shaved the hair off their backs, divided them into two groups and gave a laser treatment with a low powered ruby laser to one group. They did not get cancer and to his surprise the hair on the treated group grew back more quickly than the untreated group. That was how "laser biostimulation" effects were discovered. (Effect of laser on hair Growth of mice (in Hungarian). Mester, E. Szende, B. and Tota, J.G. (1967). Kiserl Orvostud 19. 628-631). Purpose of the work: The effects of pulsed monochromatic light, with fixed pulsations and wavelengths, on the healing of pressure ulcers were evaluated in this prospective, randomized, controlled study. Method: A placebo-controlled, double-blind study using low energy photon therapy (LLLT) was performed in ten patients with bedsore on the back.

Hemoporfin Photodynamic Therapy (HMME-PDT) is a new technology for treating port wine stain. Pain is one of the major and intractable side-effects in the treatment. We investigated the degree of pain in the patients treated by HMME-PDT and evaluated the analgesia effects of compound lidocaine cream. We found the pain during treatment started at the beginning and got worse in the first 16 minutes by time, then stabilized. Compound lidocaine cream could not release the pain during treatment.

This work presents the short-term and long-term in vivo biodistribution, pharmacokinetic and toxicology studies of novel cadmium-free indium-based QD nanoparticles (bio CFQD® nanoparticles). The pharmacokinetic studies, using inductively coupled plasma mass spectroscopy (ICP-MS) showed that the majority of QDs were accumulated in the liver and spleen and were gradually excreted from the body. Moreover, in vivo biocompatibility of QDs following systemic injection in rats were demonstrated with histopathological, biochemical and haematological assessments. Furthermore, QDs potential for in vivo sentinel lymph node (SLN) imaging was demonstrated and the optimal dose for the SLN imaging in mice was identified.

Based on the three-dimensional dispersive finite difference time domain(FDTD) method and Maxwell stress tensor equation, the optical trapping properties of nanoparticle placed on the composite metallic film are investigated numerically. Surface plasmon polaritons are excited on the metal-dielectric interface with particular emphasis on the crucial role in tailoring the optical force acting on a nearby nanoparticle. In order to obtain the detailed trapping properties of nanoparticle, selected calculations on the effects of beam waist radius, sizes of nanoparticle and circular holes, distance between incident Gaussian beam and composite metallic film, material of nanoparticle and polarization angles of incident wave are analyzed in detail to demonstrate that the optical trapping force can be interpreted as a virtual spring which has a restoring force to perform positive and negative forces as nanoparticle moving closer to or away from the centers of periodic structure.

Basal cell carcinoma mostly affects elderly people. It would be useful if the patient could receive an effective PDT treatment in a just single day, providing the same results. This study evaluated the effectiveness of this new PDT single visit protocol where the patient received all the treatment in just one visit and also the long term follow up. This study showed 93% of complete response for 220 nodular basal cell carcinoma. Kaplan-Meir survival plots showed the results from up to 33 months, 72% of disease free survival was achieved. These data allows the safe choice of single visit PDT treatment.

Breast cancer represents the most lethal type of cancer among women and its treatment with conventional chemotherapy remains a challenge. One explanation relate to enhanced efflux capacity that effectively excludes cytotoxic drugs from the cell. Hence it is significant to identify and evaluate a novel therapeutic approach. The use of a photosensitizer (PS) and light to produce death-inducing levels of reactive oxygen species in Photodynamic therapy (PDT) has the potential to meet the unmet common medical needs. In this study, we aimed to develop a resistant MCF-7 breast cancer cells (MCF-7/DOX) and study the novel photodynamic treatment efficacy. The MCF-7/DOX cells was developed by continuous exposure to increasing concentrations of Doxorubicin (DOX) treatment. The MCF-7/DOX cells were treated with Aluminum tetra-sulfonic phthalocyanine PS and irradiated at 10 J/cm2. Our results showed significant increase in cell cytotoxicity, decreased viability and proliferation.

Photodynamic therapy (PDT) is a good option for actinic keratosis (AK) treatment because it treats cancerization fields. PDT is painful, and large area treatment can be even more. This study compared the clearance of AK and pain of two PDT treatments for upper limbs (3 hours of occlusion and 1hour and 30 minutes of occlusion) using a device designed for illumination of both upper limbs at the same time. It was observed less pain with a short incubation time, maybe because the less protoporphyrin formation, but it was enough to provide the same efficacy on treatment of AK.

The purpose of this study was to evaluate the efficacy of aminopentanilic acid photodynamic therapy (ALA PDT) for female precancerous lesions of reproductive tract. A total of 210 subjects with cervical intraepithelial neoplasia (CIN, n=128) or vaginal intraepithelial neoplasia (VAIN, n=82) lesions were enrolled in the study and each received 3-6 times of ALA PDT. 186 patients (88.4%) were evaluated negative results by cytology，HPV or pathology 3 months after treatment. Three-month follow-up showed that 24 cases (11.4%) were still positive results (87.5% persistented and 12.5% progressed). 104 cases (49.5%) of HR-HPV patients were remission in the study. The study showed that ALA PDT is a promising treatment for precancerous lesions of female lower reproductive tract.

【Abstract】 The systematic nursing care was provided to the patients with condyloma who received photodynamic therapy,44 cases patients have accomplished the Systematic nursing care and compared to 44 cases patients who received routine nursing.The completion rate of thearpy in two group was 84.1% vs 68.2%(P＜0.05). The patients' satisfaction degree was 93.2% vs 86.4％ (P＞0.05). The treatment outcomes were no significant difference but the effective rate of treatment in experimental group was also higher than control group. During the 6 months follow-up, the rate of recurrence was 18.2%vs31.6% (P＞0.05).

In this study, we mainly reviews the three elements of PDT and the causes of cancer recurrence and progression after PDT. In view of the current limitations of PDT, the following questions need to be explored, to improve the stability and targeting of photosensitizers in vivo, to increase photodynamic effects and reduce phototoxicity, to overcome the hypoxic environment in the tumor, to ensure accurate monitoring and analysis of the distribution and yield of active oxygen, to analyze genic mutation status before and after PDT, and to explore whether PDT combined with other treatments could lead to new discoveries.

When the dermatologist faces a superficial basal cell carcinoma, there are many possible treatments. It would be useful if the patient could receive an effective PDT treatment in a just single day, providing effective results. This study evaluated the effectiveness of this new PDT single visit protocol and also characterized the lesion according to size, localization and recurrence after 6 months. Most of the lesions were located on trunk (57%), followed by head and neck (25%), upper limbs (14%) and lower limbs (4%). The diameter of lesions had 1.3cm in average. The 30th day biopsy showed 98.6% of complete response. After 6 months, recurrence was observed in only 4.2%. These data makes this single visit PDT treatment an excellent option for small superficial basal cell carcinoma

Sono-photodynamic therapy (SPDT) has a more obvious antitumor effect than sonodynamic therapy (SDT) or photodynamic therapy (PDT). We synthetized GPC3-targeted curcumin-loaded microbubbles (GPC3-CUR-MBs). We found that GPC3-CUR-MBs had no significant toxicity to HepG2 liver cancer cells. For the anti-liver cancer effect in vitro and in vivo, when we used CUR, CUR-MBs or GPC3-CUR-MBs as the sono/photosensitizers, the SPDT was superior than the SDT or PDT alone. GPC3-CUR-MBs were better than CUR or CUR-MBs in SDT, PDT or SPDT groups. During the treatment period, the weight of the HepG2 tumor-bearing mice did not decrease significantly, no significant evidence of lung, heart, liver, spleen and kidney damage was found in the HE staining.

We treated three different human cancer cell lines with photodynamic therapy (PDT) in combination with radio frequency electromagnetic field (RF-EMF) - mediated hyperthermia (HY) or by a conventional incubator. PDT was performed with an herbal or a synthetic photosensitizer. Afterwards cell viability was monitored and microarrays performed. PDT effects were more prominent in all cell lines by using RF-EMF mediated HY, in particular with pulse modification. A number of genes differ between conventional PDT and PDT combined with RF-EMF mediated HY, including those related to the immune system.

In summary our data provide a strong evidence indicating that variations in energy fluence rate affect differentially tumor cells displaying particular metabolic conditions and can determine the success of PDT. Indeed, the strongest proof of this interpretation is that a lower energy fluence rate can strongly modulate the fate of breast tumor cells submitted to MB-PDT. Additionally, we have also shown that these changes in energy fluence rates result in differential induction of intracellular oxidative imbalance, which is fundamental to improve the killing efficiency of the therapy.

The stability and dynamics of an arbitrary number of chiral particles subject to Bessel beam incidence are investigated. We discuss the influence of particle number and chirality parameter in detail. Linearly and circularly polarized incident Bessel beams are considered, and the corresponding BFs are compared and analyzed. It is shown that the optical BF between chiral spheres can be significantly discriminatory in nature, depending upon both the handedness of the interacting particles and the polarization of the incident light. In binding chiral spheres, therefore, the polarization of incident beams should be chosen in accordance with the chirality. This finding could be a promising avenue in controlling the optical micromanipulation on chiral structures self-arrangement.

Optical binding force (BF) exerted on stratified cells induced by a high-order Bessel beam (HOBB) is investigated with particular emphasis on the half-conical angle of the wave number components. Different types of cells, including a real Chinese Hamster Ovary (CHO) cell and a lymphocyte which are respectively modeled by a coated and five-layered sphere, are studied. Numerical effects of various parameters such as beam polarization angles, incident wavelengths, particle sizes are numerically analyzed in detail. This investigation could provide a foundation for the study of lateral BF between more complex multilayered biological particles by HOBB.

A 24-year-old female came to our clinic because of painful lesions on her face and trunk. She has been suffering from refractory aplastic anemia for 10 years. When she had taken stanozolol for 5 months, pustules, deep pustules , nodules and inflammatory sinus tract with pus appeared on her face and trunk, what’s worse is that her aplastic anemia continued to deteriorate, so she needed autologous stem cell transplantation. We used topical photodynamic therapy mediated with 5-aminolevulinic acid for her severe inflammatory acne. After five sessions, all of the inflammatory acne had been cleared to satisfy the needs of transplantation.

SUMMARY Cutaneous horn is a hyperkeratotic projection of the skin resembling the horn of an animal that have a wide range of underlying benign and malignant pathological changes. Though, benign, a cutaneous horn holds the potential to be premalignant or malignant. Therefore, it is often difficult for the excision margins of the initial surgical treatment. A retrospective, interventional case series study was performed in order to determine the efficacy of ALA-PDT for cutaneous horns and its role in surgical improvements. In total, 10 patients with cutaneous horns treated with PDT between June 2016 and June 2018 were reviewed. Clinical cured rate was 100% in all of the patients. Patients were followed up for 6 months at least, None of 10 lesions were relapsed and also had better results in terms of healing period and cosmetic satisfaction. Our study demonstrates that ALA-PDT after a shave biopsy have the potential to treat cutaneous horn in selected cases.

Topical hydrogen peroxide can induce cell death at high concentrations. In this study repeated applications of 33% hydrogen peroxide are used to reduce the size of non-melanoma skin cancers prior to excision. Small lesions are easier to excise resulting in lower costs, faster healing and reduced morbidity. From the initial six patients, a range of responses from no response to no visible lesion remaining were recorded. Margins from all excisions have been negative. Topical hydrogen peroxide may be an inexpensive and effective method to reduce the size of non-melanoma skin cancers.

Photodynamic diagnosis and therapy is an interdisciplinary subject which integrates traditional and state-of-the-art, In order to establish a simple and universal solution, the idea of computational microimaging is used adequately on global optimization, system reconstruction and complex structure control, which overcome the shortcomings of complex relationships among time series, spatial distribution and factor control, and play a significant role to improving in micro-environment. As the criterion group are also effectively considered, comparative study can demonstrate that the dialectical relationship and their balance is important, especially among Merit function, Optimization algorithms and Model parameterization. The results can develop new products.

PDT requires of efficient photosensitisers and of selective drug delivery strategies to target the optimum cellular localization within the body and furthermore, into their optimal organelle, so to enhance the treatment outcome. For this purpose, 2,7,12,17-methoxyethylporphycene has been conjugated to an antibody, an antibiotic and a lipophilic cation, namely, trastuzumab, gentamicin and triphenylphosphonium. An in-depth photophysical study and biological in vitro assays of the conjugates have been performed to evaluate their optical properties and the phototoxicity induced to eukaryotic and prokaryotic cells, examining the selectivity achieved.

For treatment-resistant Vulvar lichen sclerosus(VLS),PDT represents a valid therapeutic option.There have been reports of laser therapy as an effective treatment option of VLS.A 70-year old woman who has a history of refractory VLS was firstly treated with ALA-PDT,the same protocol was repeated for 3 times at a 2 week interval,then Holmium laser and ALA-PDT were combined to treat the same areas at a time.Satisfactory results were noted without any unbearable adverse effects.The combination of Holmium laser and ALA-PDT could be an effective treatment option with good tolerance for VLS patients who are refractory to other standard therapies.

Head and Neck Squamous Cell Carcinoma is the seventh most frequent cancer worldwide, its treatment is limited by tumor recurrence and metastasis, even with advances in traditional techniques. This scenario emphasizes the importance of non-traditional treatment techniques such as PDT. Photobiomodulation is based on applying light to modify cellular processes such as proliferation and tissue regeneration. Therefore, we investigated whether PBM contributes to the tumor regression by PDT due to the increase of the concentration of ROS in treated cells as well as promoting the absorption of photosensitizer by the tumor cells.

Objective: To observe the efficiency and safety of hematoporphyrin mono-methylether photodynamic therapy (HMME-PDT) in treating port-wine stains (PWS) with Chinese patients, and to evaluate the advantage of photograph、VISIA-CRTM system, and dermoscopy in efficacy evaluation. Method:52 patients were treated in our hospital over the past two years with HMME-PDT ,among which, 15 cases were pink type, 29 cases were purple type and remain 8 cases were Nodular thickening type. Initially, All patients received an intravenous injection of 5 mg/kg HMME ,and then the lesion areas of the patients were exposed to 532 nm LED green light after 10 minutes. The irradiation power density was range between 80–95 mW/cm2. By utilization of photograph、VISIA-CRTM system, and dermoscopy to evaluate the clearance after treatments, and then informing the patients to rank the pain level during the treatment via VAS scale, and recording the side effects.Result:After 1-7 times treatments.13 of the 52 cases were cured (25.00%) , 14 cases showed a good efficacy (26.92%), 16 cases indicates alleviation (30.77%), while 9 cases displayed no efficacy (17.31%).By observation, it showed that the pain level each patient could endure were distinct, and it’s remarkable that when receiving consecutive 11-12min of treatment, most of patients have showed with severe pain (according to VAS scale). The side effects after treatment mainly displayed with edema，crust，hyperpigmentation.Conclusion: it shows that after treating with HMME-PDT ,the efficacy is remarkable and with advantage of safety and less side effects, which worthwhile for further research and promotion.

A 46-year-old woman presented with a 6-month history of a recurrent lobulated nodule involving her perineum that had first appeared 6 months previously. Diagnostic biopsy specimen confirmed Bowen’s disease. Topical 5-aminolevulunic acid photodynamic therapy（5-ALA PDT） with was administered 6 times, over the course of 3 months. The patient was seen every 3 months after treatment. Twelve months after treatment, no clinical evidence of the condition was observed, with preservation of a normal perineum. 5-ALA PDT represents an effective strategy for treatment of Bowen’s disease, with structural and functional preservation of the involved organ.

This study compared the efficacy of photodynamic therapy (PDT) combined with carbon dioxide (CO2) laser treatment, PDT treatment and CO2 laser treatment in the treatment of condyloma acuminatum (CA) in anal tube of patients with HIV. The recurrence rate of PDT combined with CO2 laser treatment group at 3 months after treatment was the lowest. Photodynamic therapy combined with CO2 laser could be a good option for the treatment of CA in anal tube of patients with HIV.

Antibody-based near infrared photoimmunotherapy (NIR-PIT) is an attractive strategy for tumor treatment. GPA33 is overexpressed in most colorectal cancers and has been verified as an attractive theranostic target. We previously produced the single chain fragment of a variable antibody against GPA33 (A33scFv antibody). Here, we investigate the efficacy of NIR-PIT by combining A33scFv with the photo-absorber, IR700DX. In vitro studies showed that A33scFv-IR700-mediated PIT could rapidly and specifically kill GPA33-positive LS174T cells. Moreover, reactive oxygen species were accumulated in LS174T cells treated with NIR-PIT. In mice bearing LS174T tumor grafts, the NIR-PIT group had significantly smaller tumors than other control groups. Thus, the A33scFv-IR700 may be an alternative antibody-photosensitizer conjugate for NIR-PIT.

In this study, we demonstrated ROSED in an ongoing Phase II clinical trial of Photofrin-mediated PDT for pleural mesothelioma. We measured light fluence rate and PS concentration using isotropic detectors sutured on the pleural cavity wall and connected to a PDT dose dosimeter. Tissue blood flow was monitored using DCS with a contact probe sutured adjacent to an isotropic detector. Different dose metrics, including light fluence, PDT dose and [ROS]rx calculated based on light fluence, photosensitizer concentration, and relative blood flow were compared.

Photodynamic therapy (PDT) induced by protoporphyrin IX (PpIX) has been widely used in treatment of skin cancers. Clearance rate depends on different factors such as light irradiation, skin oxygenation and drug penetration. The penetration of 5-aminolevulinic acid (5-ALA) with topical application is currently limited and restrains the production of PpIX which could restrict PDT outcomes. A microneedle (MN) patch containing 5-ALA has been developed to accelerate drug delivery and enhance PpIX presence in deep skin layers. After an in vivo experiment on hairless rats, MN-patch’s parameters have been optimized in order to obtain maximum PpIX fluorescence.

The results of PDT with local application of the 12% Levulon gel indicate its high efficiency in combination with high cosmetic results. This type of treatment is recommended primarily to patients with superficial BCC with the tumor foci localization on the skin of face.

This study assessed the efficacy of autofluorescence endoscopy (AFE) using the Onco-LIFE system and numerical color value (NCV) estimation in comparison to white light endoscopy (WLE) in endoscopic surveillance for identification of early dysplasia in Barrett’s esophagus (BE). In the case of dysplasia photodynamic treatment (PDT) was carried out. In the case of 248 biopsies taken from sites with NCV below 1.0, two cases of unspecified dysplasia were recognized; in 14 biopsies with NCV above 2.0 in all cases the various grades of dysplasia were documented. Dysplasia was found in 42% of AFE+NCV- guided biopsy specimens, and in 7.1% of WLE-guided biopsy specimens. AFE+NCV detected high-grade dysplasia in 7 patients, 6 more than according to SP in WLE. In the group patients with dysplasia, PDT was successfully carried out.

Background: Pulsed dye laser (PDL)-resistant port-wine stain (PWS) is difficult to manage. Objective: To assess the efficacy and safety of vascular-targeted photodynamic therapy (PDT) in patients with PDL-resistant PWSs. Methods: We retrospectively analyzed 70 patients with PDL-resistant PWSs who were treated with vascular-targeted PDT. Its clinical efficacy was assessed by a chromameter and visual assessment of color blanching and size reduction of the PSW lesion. Adverse events and patient satisfaction were evaluated. Results: The median blanching rate of the PWS lesions was 29.81% after an average of 1.56 sessions of PDT. Overall, 47.2% of patients had excellent or good improvement, and 51.4% achieved >25% reduction of the lesion size. Adverse effects were transient and self-limiting. Conclusions: PDT is promising for treating PDL-resistant PWS. The anatomical location of the PWS lesions is an important predictor of the patient’s response to PDT.

The purpose of our study was the evaluation of the practical usefulness of endoscopic autofluorescence assessment (AFE) using the Onco-LIFE system, and the estimation of the correlation between the histopathological evaluation with the degree of lesions’ Numerical Color Value (NCV index) and the method’s sensitivity and specificity valuation. In the group of 67 patients, we found 44 cases of primary or secondary cancers and 7 cases of non-epithelial malignancies. In this group (51 patients) we identified 13 colorectal cancers and 38 upper gastrointestinal cancers. Based on the NCV index at NCV>1.0, we can show that the sensitivity for malignant neoplastic lesions was 100% and the specificity was 73%, while for NCV>1.5, the sensitivity for malignant neoplastic lesions was 86% and the specificity 100%.

Although fluorescence-guided surgery using the cancer-specific accumulation of PpIX is used to identify tumours during brain tumour surgeries, improvements in the technique are required for increasing its efficacy. In an earlier study, we discovered that inhibiting the oncogenic Ras/MEK signalling pathway enhanced the cancer-specific accumulation of PpIX in colon and breast cancer. Here we investigate whether MEK inhibition would enhance PpIX accumulation in brain cancers in vitro and in vivo. We also developed a MEK inhibitor-loaded nanoparticle with high affinity to glioblastoma cells and is capable of crossing the blood-brain barrier, to improve intraoperative visualisation of brain tumours.

Daylight PDT is a preferred treatment for field-change actinic keratosis. During dPDT, a patient should receive a minimum threshold effective light dose to ensure effective treatment. However, light attenuation by modern sunscreen formulations is not currently accounted for, and it is hypothesised that the increased protection at longer wavelengths offered by modern sunscreens may interfere with activation of PDT effects by daylight. By analysing the transmission spectra of several sunscreens, we show that effective dPDT dose is indeed reduced by a significant amount by sunscreen application, and that practitioners of dPDT should be aware of these findings.

Cutaneous leishmaniasis (CL) is a neglected disease, which promotes destructive lesions. Difficulties in treatment are related to resistance and toxicity. Methylene blue-mediated photodynamic therapy (MB-PDT) has emerged as a promising treatment considering low cost and no reports about resistance. We evaluated MB-PDT on Leishmania amazonensis, in vitro and in vivo using a red LED at different fluences. Our results demonstrated that the best fluence in vitro was not effective in vivo, and a higher dose was necessary to provide better responses mice. This study reinforces the idea that a well-planned protocol is necessary for a successful MB-PDT on CL.

The treatment of disseminated cancers such as peritoneal carcinomatosis (PCAR) involves chemotherapeutic regimens that are highly toxic and poorly tolerated by patients. Designing minimally toxic treatment strategies is therefore imperative. Taking advantage of tumor-specific metabolic pathways through targeted therapeutics and diagnostic agents therefore represents a promising treatment strategy, for which a better knowledge on cancer metabolism is necessary. In this presentation, we present the characterization of metabolic heterogeneity and plasticity of patient-derived organoids of PCAR, which guides the development of metabolically-enhanced theranostic treatment modalities such as photodynamic diagnosis and therapy.

In dermatology, photodynamic therapy (PDT) relies on the photo-activation of the photosensitizer protoporphyrin IX (PpIX). We have previously developed a freely available website (http://www.oncothai.fr/light-efficiency-calculator/), which computes the PpIX-weighted irradiance of any uploaded spectral irradiance. Resulting from the weighting of this spectral irradiance by the normalized PpIX absorption spectrum, the PpIX-weighted irradiance allows to quantify the efficiency of the corresponding light source in photoactivating PpIX. Through this website, we have assessed a variety of light sources proposed for use in dermatological PDT and obtained a wide range of PpIX-weighted irradiances with a maximum value for daylight on a clear sunny day.

Photodynamic Therapy (PDT) is showing great potential in skin cancer treatment. Recently, new protocols, different light sources, and photosensitizers are being tested in order to improve PDT efficiency. Although PDT response evaluation is related to treatment follow up, predict PDT response is important while studying protocols. Imaging in vivo tissue auto-fluorescence lifetime this work aims to investigate PDT treatment response. Endogenous metabolic contrast lifetime images of basal cell carcinomas (BCC) after six months of PDT treatment were studied. Preliminary results show differences on fluorescence lifetimes between residual BCC and complete response after PDT treatment.

We conducted PDT on patients with oncological diseases and selected exudate from tumor tissue. Then, the exudate was checked for the lauch of apoptosis and was applied on surface of tissue culture flask that contained systematically crossed cancer cell culture. Exudate from a human tumor, in which cells with PDT-induced apoptosis are located, starts the process of apoptosis in most of the malignant cells in systematically crossed cancer cell cultures. The data that was obtained show a new way to combat malignant neoplasms and metastases based on the tumor apoptosis factor (TCApF) induced by PDT, and allow us to create a new type of “vaccine” against cancer.

Vessel recovery is observed some hours after photodynamic therapy and can be an angiogenic process or blood reperfusion. For the investigation of this vascular process after photodynamic therapy, the chorioallantoic membrane (CAM) was used with 10 μg/mL of photosensitizer Photogem® and subdoses of light. For quantification of these effects, an equation was determined, routine of MATLAB® was used to determine the percentage of blood vessels area and an analysis of the distribution of large and small vessels was performed with ImageJ®, showing that smaller vessels are most affected 3 hours after the therapy, recovering the number after 24 hours.

The unpredicted partial responses of PDT are a clear signal of the necessity for a monitored application of this therapy. To overcome the shallow penetration of the most used spectral range for PpIX fluorescence excitation (around 405 nm), we developed a system to both treat and monitor PDT using the NIR PpIX fluorescence emission (around 700 nm). The system uses the 633 nm treatment light itself to excite PpIX fluorescence emission in the NIR region, which is imaged in real time by a camera. The system was validated in clinic for the monitored treatment of skin lesion with promising results.

Photodynamic therapy (PDT) is an effective treatment modality in a variety of solid cancers due to the lack of cumulative toxicity and slight mucosal damage. In this study expression of biomarkers Ki-67, EGFR, p-EGFR, CD31 and CD44 before and after PDT was examined for 18 patients with histologically confirmed T1N0M0 micro-invasive squamous cell carcinoma of the buccal mucosa treated with Aminolevulinic acid (ALA)-based PDT. The immunoreactivity post PDT decreased in intensity and proportion in all the cases except 4 cases with recurrence of the disease. Immunohistochemical expression of biomarkers can help in the prognostification of PDT treated micro-invasive cancers.

Blood, considered a highly nutritive medium, can be the target of microorganism contamination. Blood bags containing erythrocytes, platelets, and plasma, used for transfusion, are also targets of contamination and can trigger serious diseases, especially blood infections. Some optical techniques may be used in the microbiological control of blood. In this study, PDI and UV radiation were evaluated in the in vitro decontamination of whole blood, erythrocytes, and platelet-rich plasma with S. aureus. The characteristics observed when PDI and UV were applied in blood and its components are presented, and results allow setting limitation and use conditions for techniques in blood.

Optical Coherence Tomography angiography can collect the information of blood vessels such as diameter and density. Moreover, photodynamic therapy is able to treat vascular disease such as port wine stain and malignant tumors through destroying blood vessels. In our study, we used the tumor which planted on the mice ear as our objective. All the mice ears with tumors were treated by PDT, additionally, applying OCTA to monitoring the vascular changing after PDT instantly, six hours, twenty-four hours，three days，five days. We found that the disappearing of vessels was associated with the effect of PDT, more obvious declining of vessels in the tumor, the volume of tumor was reduced more significantly.

Hyperspectral (HS) imaging systems have become important tools in an array of fields, but certain methods for spectral unmixing present a significant computational bottleneck in the HS imaging pipeline. This work uses GPUs to accelerate a method for real-time hyperspectral decomposition via a per-pixel non-negative least squares fitting routine that accommodates a wide range of spatial, spectral, and temporal resolutions. This algorithm achieves video-rate (> 15 fps) analysis for a range of HS data sets at an average throughput of 2.5 GB/s. The method is also applied to a hyperspectral fluorescence imaging system to show online 5-color optical biopsy (5 protein biomarkers) in a mouse model of ovarian cancer to monitor responses to PDT.

Literature shows a growing interest in using optical techniques for microbiological control. In transplantation procedures, the availability of organ donors is often reduced, and several times organs that are compatible and viable are dismissed due to contamination by pathogenic microorganisms. Our study aims to offer an alternative for decontamination of transplantation organ grafts by a “mechanical plus photophysical” process, using either ultraviolet light or photodynamic activation for decontamination of a circulating preservation solution during cold perfusion stages of transplantation procedures. Early results show reduction between 3-4 log for Staphylococcus aureus tests in PBS and in preservation solution for transplantation procedures.

The development of hyperspectral microendoscopes for pre-operation optical biopsy to quantify biomarker targets for precision fluorescence guided surgery and photmedicine provides motivation for adapting mosaicking algorithms to process a plurality of simultaneous hyperspectral channels. We present an algorithm that mosaics hyperspectral microendoscopic video by correlating channels of consecutive frames as a basis for calculating image alignments. Furthermore, we employ parallel processing via GPUs to alleviate computational bottlenecks and approach video-rate mosaicking speeds. This implementation lays the foundation for real-time multi-channel mosaicking to accompany video-rate hyperspectral microendoscopic probes.

Chromoblastomycosis is a rare fungal infection involving skin tissues, usually presenting as partial hypertrophic and warty plaque. Effective treatment is necessary to control the development of lesions, especially in patients with associated diseases. Photodynamic therapy (PDT) is an efficient and non-invasive treatment option. We reported the case of a 52-year-old male with refractory chromoblastomycosis and leukopenia, who was successfully treated with 5-aminolevulinic acid-based PDT (ALA-PDT). A complete cure, confirmed by clinical improvement and mycological detection was achieved after six sessions of every-other-week treatment. Post 6 months follow up no recurrence was observed. The case here suggests that ALA-PDT is a valuable therapy for refractory chromoblastomycosis.

Fiber optic scanning microendoscopy enables fluorescence microscopy deep within the body. These devices are essentially miniature laser scanning microscopes for linear (confocal or wide-field) and nonlinear (multiphoton) imaging applications. We present simple methods to fabricate a low-cost miniature fiber scanning microendoscope probe with specifications that promise video rate imaging applications.

The incidence of dengue (DEN), chikungunya (CHIK), and Zika (ZIK) has grown dramatically in recent years. In Brazil, for example, dengue infected around 47.393 people in 2018, causing approximately 3.341 serious cases. Photodynamic inactivation (PDI) through the use of natural photosensitizers (PS), such as Curcuma longa derivatives (Curcumin, Demethoxycurcumin and Bis-Demethoxycurcumin), has proved to be a promising technique to eliminate Aedes aegypti larvae in breeding sites (the main vector for DEN, CHIK and ZIK). This study shows original and relevant results of the high efficacy of curcumin photolarvicide properties against Ae. aegypti larvae, under field conditions.

Fiber lasers offer promise as a means to produce ultrashort pulses of light suitable for nonlinear microscopy in compact, robust and portable devices. We will present a protocol to fabricate a low-cost all-normal-dispersion ytterbium (Yb)-doped femtosecond fiber laser oscillator using commercially-available parts as well as basic fiber splicing and laser pulse characterization equipment. Methods to verify true versus seemingly (partial or noise-like) mode-locked performance will also be presented to illustrate the intellectual investment required to build a custom femtosecond fiber laser. These approaches make custom fabrication and routine use of femtosecond fiber lasers more accessible to a wide range of investigators.

Bacterial colonization and biofilm formation on catheters are the primary cause of nosocomial urinary tract infections, entailing a limitation for the long-term use. We have developed self-sterilizing silicone catheters with a covalently-attached layer of photosensitizer (PS) that showed strong antibacterial effects in vitro against a panel of bacterial species commonly associated with catheter-related infections. Exposure of the PS-derivatized catheter to 532 nm laser light induced killing of more than 96% of the bacterial population within 30 min of illumination. Biofilm formation by Pseudomonas aeruginosa or Staphylococcus epidermidis was prevented on the PS-tethered catheters compared with underivatized controls.

The use of photodynamic therapy (PDT) in cancer treatment has shown remarkable efficacy when used in combination with other approved therapies. However, the large parameter space to be explored when implementing combination treatment with PDT can be daunting. A challenging aspect of this design is the timing of treatment administration, as this requires longitudinal measurements of cancer cell behavior. Repeatable, in-vivo molecular imaging with cellular resolution would allow detailed knowledge of cancer response to targeted therapies and may identify temporal windows for optimal treatment. Here we report recent results demonstrating multicolor microendoscopy as a new diagnostic for in-vivo molecular imaging.

Photodithazine® (FS) photosensitizer interaction in Candida albicans (CA) applied to photodynamic therapy. Temporal and spectral experiments using confocal fluorescence microscopy were performed to determine the uptake of FS by CA, demonstrating internalisation by the excitation light for FS. Induction performed in low doses of light radiation of FS in large areas. Excitations with diffused light by LEDs (660 nm). Cell viability with applied doses of continuous and serial light. Serial application promotes greater FS entry in the AC cell and greater cellular lethality compared to the single dose of the same value.

Daylight photodynamic therapy (dPDT) is an effective and commonly used treatment for actinic keratoses, which are caused by chronic ultraviolet (UV) exposure. Using historical data and a two-hour exposure, the standard erythemal dose (SED), UVA dose and dPDT dose were calculated and compared. Peak UV exposure occurs around solar noon, during the summer and is higher at lower latitudes. A minimum threshold dPDT dose can be achieved at times of the day and year when UV exposure is lower. These data may provide useful information for clinicians to aide administration of dPDT during periods of lower UV exposure.

Control of pathogenic microorganisms from peri-implant pockets is one important step to treat peri-implantitis. 20 implants were randomly into two groups: I) received conventional treatment; and II) group PACT received conventional treatment and PACT mediated by PapaMBlue® and red laser. We have used laser device (Therapy EC, DMC, São Carlos, Brazil) emitting wavelength of λ=660 nm and out-put power of 100 mW for 2 min at each site, 30 J/cm2 radiant exposure and power density I=250 mW/cm2. We have collected microbiological samples of both groups before and immediately after treatments. Data showed that PACT decreased bacteria boarder in 1 log.

Healthcare-associated infections (HAI) are important life-threatening conditions, and the link between oral cavity health status and HAI is of great importance. Periodontal disease is a common form of oral infection and its relationship with HAI is of great interest nowadays. In this study we present the importance of aPDT on periodontal disease treatment regarding the control of pathogens connected with HAI. According to our results periodontal treatment associated with aPDT, performed with methylene blue and a 660nm diode laser, can improve clinical aspects of patients with chronic periodontal disease, and the compromised sites, that were colonized by HAI related pathogens, one week after treatment were free of HAI related pathogens.

Ventilator Associated Pneumonia (VAP) is one of the most dangerous respiratory disease that can be caused by Staphylococcus aureus biofilm on endotracheal tube. Almost 69% of the pacients with the endotracheal tube developed VAP at Intensive Care Unit (ICU). This complication increased the permanency these pacients in the hospital. The treatment for VAP have been a challenge due to the bacterial resistance to antibiotics. The photodynamic therapy using curcumin and blue light were chosen for inactivated the S. aureus biofilm at the endotracheal tube.

Oral cancer represents over 30% of cancers reported in low middle-income countries. Surgery, radiation and chemotherapies are current modalities which are expensive and have side effects. We combined engineering, optics and biochemistry to produce low-cost, mobile LED-based light source with 3D-printed light applicators for smart phone-based, photodynamic therapy. After validating the devices in preclinical models, we performed an ergonomics study on 10 healthy volunteers, where the comfort level of the applicators was evaluated. Further, the device was tested in clinical studies of early oral cancer in India. This study offers an alternative treatment modality for early disease without associated morbidities.

Determination of in-vivo tissue optical properties for anal PDT is challenging due to the light integrating-sphere effect in an enclosed cylindrical cavity. In this study, we developed an analytical model to calculate for scatter light fluence rate and optical properties based on measurements of light fluence rate from an enclosed cylindrical cavity. A special anal PDT light applicator was used to performed measurements in tissue mimicking liquid phantoms. The agreements between measured and calculated scatter light fluence rate on the surface of the cylindrical geometry and the optical properties will be compared.

Biofilm matrix blocks the diffusion and prevent the action of a variety of molecules, including photosensitizers used for antimicrobial photodynamic therapy (aPDT). We explored pH to overcome penetration issues and optimize aPDT. Staphylococcus aureus was grown in biofilms and treated with methylene blue (MB; 50 or 100 µM) in different pH (4, 5, 6 or 7.4), following irradiation (657 nm LED; 50 J/cm2) and resazurin assay. Acidic aPDT was more effective in reducing biofilm viability than aPDT in neutral pH. Further investigation is required but controlling pH might be the key to achieve complete eradication of biofilms with aPDT.

Cutaneous leishmaniasis is a neglected disease, with a treatment aggressive, and Photodynamic therapy is a promising treatment. This study evaluated the damages triggered by PDT with porphyrin Leishmania promastigotes. Porphyrin was localized in cytosol and flagellum, Viability was not affected in groups treated in the dark, however, post PDT was observed only 20% of viability in the groups treated in the highest concentrations. There was morphological alterations pos pdt, yet, seems that tubulin structure was not completely disrupted. These results suggest that porphyrin may be a promising photosensitizers to be used in the PDT treatment of cutaneous manifestations of leishmaniasis.

We evaluated the potential of electromagnetic (EM) tracking for interstitial photodynamic therapy (I-PDT) light source placement. Treatment planning for I-PDT defines light-source positions for effective PDT. Accurate light-source placement to these positions is necessary to ensure correct light dose and light dose-rate. Plastisol phantoms containing targets were constructed and catheters containing graphite were inserted at each expected target location using EM tracking and ultrasound. CT images post-graphite injection revealed graphite was accurately injected at target sites in opaque phantoms using the EM tracking system. We propose utilizing EM tracking to assist with light-source placement for I-PDT.

Dosimetry during photodynamic therapy has been one of the most significant challenges in the application of this promising cancer therapy. Real-time feedback on the distributions of the photosensitizer and singlet oxygen during treatment will be a valuable tool for PDT researchers and clinicians. Accessibility to this information could lead to much higher efficacy of clinical PDT treatments. Physicians would be able to personalize light doses for PDT adapting to the different treatment responses of individual patients.

Aspergillus fumigatus is the most common isolated agent in chronic invasive fungal rhinosinusitis with patients enduring painful and recurrent sinus symptoms. Prompt diagnosis and initiation of appropriate therapy is essential to avoid a protracted or fatal outcome. Our group has developed a novel methylene blue-based photodynamic therapy approach to treatment of Aspergillus rhinosinusitis. Briefly, A. fumigatus was encapsulated within 250μm agar beads in a validated model and titrated into the rabbit maxillary sinus. A. fumigatus burden after Sinuwave™ antimicrobial photodynamic therapy was compared to control, with > 99.9% reduction in recoverable fungus, a highly significant outcome.

The effects promoted/observed during the interaction of light with the most varied biological tissues make it an interesting tool both for diagnostic and therapeutic purposes. Monte Carlo simulations are considered important and reliable tools for studies on light propagation in tissues. The aim of the present study was to use Monte Carlo eXtreme to predict possible changes in light propagation when adding a layer of transparent material between the air and the turbid medium. The simulations performed in this work demonstrated that the addition of material between the air and the phantom modifies the shape of the light distribution.

Our objective was evaluating its photodynamic inactivation of bacteriochlorin on Candida albicans. The bacteriochlorin was synthesized from the extraction of bacteriochlorophylls from non-sulfurous purple bacteria and then converted to bacteriochlorin. For the experiments, the Candida albicans were standardized at the concentration of 10^6 CFU/mL. Different treatments were applied to the culture (light doses associated with varying concentrations of photosensitizer). The quantitative evaluation of viable cells was performed by plate counting using the spread plate method in Sabouraud Dextrose Agar, incubating at 36±1ºC for 24 hours. The results showed that the photodynamic inactivation for Candida albicans was considerable; the treatments reduced cell viability maximum 2 logs.

Photodynamic therapy (PDT) combines light, a photosensitizer, and molecular oxygen in tissue to kill malignant diseases. Normally, ALA is administered orally or topically and is preferentially retained by the target tissue, which is subsequently exposed to light at 630 nm. We have performed an ALA-PDT in mice bearing radiation-induced fibrosarcoma (RIF) tumors on the shoulder to determine the photochemical parameters necessary to determine reactive oxygen species concentration ([ROS]rx) during PDT.

Since the 1990s, the number of published articles carrying the keywords [“antimicrobial photodynamic” OR “photodynamic antimicrobial”] has been increasing exponentially, mainly from 1998 on. Brazil accounts for 26% of all aPDT publications in the world, 55% of the Americas and 95% in South America, contributing extensively to unveil molecular mechanisms, developing protocols for oral, dermatologic and veterinary infections, and to improve the technique so it can become a clinical reality. In this presentation, we cover the numbers of South America’s research and provide a full network list to encourage scientific collaborations.

In this study, we use Monte Carlo modelling to investigate the effect of tissue optical properties on light scattering inside an enclosed cavity surrounded by tissue. MC simulations are performed for absorption coefficients between 0.1 – 1 cm-1 and reduced scattering coefficient between 2 – 40 cm-1. The MC simulation codes are based on either an in-house code for spherical and cylindrical geometries and the open source mmc (Mesh Monte Carlo) code modified to allow multiple scattering inside the non-scattering cavities.

We report incorporating finite-element-based light propagation simulations with photokinetics of the light-activated drug (PS) to create a dosimetry system for light fluence rate, fluence, and induced fluorescence, in near real-time and in 3D, for PDT treatment in the clinic. Recent experiments in phantoms at Roswell Park Cancer Institute show excellent agreement with calculations. Future efforts will develop a novel hardware and software prototype platform with feedback and monitoring that integrates real-time light and PS dosimetry.

Onychomycosis is a nail disease caused by fungi, and isthe treatment is difficult, which consists of the antifungal agent’s administration. Due to the increase of drug-resistant microbial strains and the high incidence of this infection, it is necessary the development of new technologies, such as photodynamic therapy. The purpose of this study is to implement PDT as a technique for the treatment of onychomycosis using a Brazilian curcumin as photosensitizer (PDT Pharma, Brazil) besides to suggest new diagnostic tools using thermographic and fluorescence imaging. A clinical study was carry out for the treatment of 50 patients with onychomycosis using a specific device for this application. The low cost, the simple operation with fast clinical results are important factors for the implementation of this technology in the treatment of onychomycosis.

Methicillin resistant Staphylococcus aureus (MRSA) is a concern to public health. Here we report a dual-wavelength combination therapy approach, using 460 and 405 nm light, that exploits the STX photo-bleaching effect of 460 nm light to sensitize MRSA to 405 nm aBL. Four MRSA strains including ΔctrM mutant (non-STX producer) were exposed 180 J/cm2 of 460 nm light immediately before exposure to 108 J/cm2 405 nm aBL. Findings demonstrated that pre-exposing wild-type strains to 460 nm wavelength significantly potentiated the antimicrobial effects of 405 nm aBL (P<0.001); suggesting this combination therapy may offer a novel approach to treating MRSA infections.

Oral leukoplakia is one of the most common oral potentially‐malignant disorders with complex causes, a long disease course and a high tendency for recrudescence. Although a variety of methods exist for treating this disease, canceration rates remain high. Herein, we described a case of 72-year-old male patient with oral leukoplakia of the palatine mucous membrane who had achieved complete remission after being treated with five sessions of plum-blossom needle assisted 5-aminolevulinic acid-photodynamic therapy. The patient had since been subsequently placed under close observation (>12 mo). To date, there has been no recurrence. Plum-blossom needle assisted photodynamic therapy might be suitable for the treatment of oral potentially‐malignant disorders in patients presenting with epithelial hyperkeratosis.

In this study, Cylindrical Diffusing Optical Fiber Probe (CDOFP) with optimal shape for tumor infiltration is suggested and self developed laser scribing equipment is used to develop and analyze 5mm~40mm diffusing length CDOFP. CDOFP was infiltrated inside a tissue to process laser coagulation test. Coagulation and thermal damage was measured with twice the maximum intensity of laser, where maximum intensity in-vivo test is 1W 200J. The CDOFP design depending on tumor's shape and size will be analyzed by the comparative test with the existing probes in various cases and then, the best CDOFP design will be proposed.

Methicillin resistant Staphylococcus aureus (MRSA) is a threat to public health, with limited treatment options available. Here we report a novel anti-MRSA combination therapy using antimicrobial blue light (405 nm) and pyocyanin (aBL+pyocyanin) that effectively bleaches STX and results in the elimination of MRSA by aBL. aBL+pyocyanin effectively eliminated MRSA in vitro, significantly potentiating antimicrobial effects when compared to either monotherapy (P<0.001); which was attributed to a powerful photo-induced chemical bleaching effect by aBL+pyocyanin. These findings support the notion that aBL+pyocyanin combination therapy, if fully optimized and developed, may be an effective strategy to eliminate MRSA infections.

Moraxella Catarrhalis is a major pathogen of otitis media (OM) in childhood and chronic obstructive pulmonary disease in adults. This pilot study aimed to investigate the potential of antimicrobial blue light (aBL), an innovative non-pharmacological approach, for M. catarrhalis infections. In planktonic suspensions, an exposure of 216 J/cm2 aBL (405nm, 60 mW/cm2) resulted in 4 to 6-log10 CFU reduction in ATCC strains and clinical isolates, while in biofilms, over 2-log10 CFU reduction was achieved. Protoporphyrin IX and coproporphyrin were the most abundant endogenous photosensitizing chromophores in M. catarrhalis. Our data suggested that aBL is a promising approach against M. catarrhalis.

Laser diodes have been widely used as monochromatic light source in photodynamic therapy (PDT). Here, we discuss the fabrication of a light emitting diode (LED) array to realize a low-cost, simple and customizable PDT setup. The maximum light intensity that reaches the biological sample plane is discussed as well as the spectrum and the stability under conditions of practical use. We offer this development as a viable alternative to comparatively expensive lasers for light delivery in PDT applications. Finally, a protocol for assembling a low-cost electronic control circuit is described to automate the sample exposure time without the need for a mechanical shutter.

This review summarizes the available data related to programmed cell death already published for the cattle parasite T. foetus and attempts to clarify some crucial points to understand this mechanism found in non-mitochondriates parasites. T. foetus can choose among different pathways how to initiate cell death. Thus, a major challenge for cellular death research remains the identification of the molecular cell death machinery of this protist and the prospects in the future research. Although, the possibility of the existence of different pathways to cell death in trichomonads is discussed and a model for possibles executioners pathways during T. foetus cell death is proposed.

Antimicrobial Photodynamic Therapy (aPDT) has been investigated as an alternative method for the inactivation of microorganisms. However, this treatment has a reduced effectiveness when the microorganisms are organized as biofilm. Recently, Sonodynamic Therapy (SDT) has also been suggested as an antimicrobial treatment and has the advantage of activating photosensitizer by the use of ultrasound (US), which propagates deeper into the tissue and is able to disrupt the biofilm. Thus, this study demonstrated that the association of US with aPDT mediated by curcumin, exhibits a higher and significant effect against Staphylococcus aureus biofilms than SDT and aPDT.

Aim: Observed the therapeutic effectiveness of Photodynamic therapy (PDT) on Vaginal intraepithelial neoplasia (VaIN). Methods: 30 patients was irradiation with 630 nm laser (100 mW/cm2, 20min) on vagina 72 h after intravenous injection of Hematoporphyrin derivative (3mg/kg). Results: 19 (63.33%)patients had histologically confirmed normal. 14 (46.67%) patients had normal Pap smear results 3 months later, and 24 (80.00%) patients had normal results 1 years later. 23 (76.7%) patients retained HPV infection3 months later, while 1 years later 15 (50.0%) patients retained HPV infection. The multivariate analysis showed that only the age was a significant variable predicting disease recurrence (P= 0.0151).

Plasmonic photothermal therapy has been recently suggested as one of the potential treatment for cancer therapy with various gold nanoparticles that can generate destructive heat inside the tissue. However, photothermal properties of the gold nanoparticles varied with the laser power, optical density of the particles, or depth of the texture, used for clinical treatment have not been established due to the insufficiencies of the heating regulation and temperature measurement techniques, leading to unreliable and inaccurate treatment. In this study, the heat generation of the nanoparticles was quantitatively characterized with all variable control for clinical treatment with breast cancer tissue model.

Zinc Phthalocyanine is a type of powerful photosensitizer with high yield of singlet oxygen generation. A number of phthalocyanine-type photosensitizers are currently in clinical use or in trials for anti-tumor applications. As an example of antimicrobial application, we synthesized a series of PSs with different positive charges (ZnPc(Lys)n, where n = 3, 5, 7, and studied their antibacterial activities and mechanisms against E. coli, including binding kinetics, bacterial surface hydrophobicity, zeta potential of bacteria, membrane permeability and bacterial signaling pathways. Our results clearly demonstrate that aggregation state of the photosensitizer on bacterial surface is a key parameter determining the antibacterial efficacy.

Recently, anti-PD-1/PD-L1 immunotherapy has shown promising clinical results. However, there is still a large number of patients respond poorly, therefore more potent combinational therapies are required. In this context, immunotherapy based on the modulation of immune checkpoint molecules alone, or in combination with PDT, is now in focus as a means of developing new therapeutic strategies for more resistant cancer. Herein, using optimized PDT protocols we design novel experimental strategy based on the local effect of Redaporfin-PDT with systemic inhibition of PD-1/PD-L1 immune checkpoint, that can result in strong antitumor response - essential for the therapeutic effect of combinational treatment.

Antimicrobial photodynamic therapy (aPDT) has been proposed to cope with the increasing antibiotic resistance among pathogens. We evaluated the antibacterial effect of sinoporphyrin sodium (DVDMS)-mediated aPDT on bacteria and MDR-bacteria in vitro and in vivo. Among the strategies to enhance the efficacy of aPDT against Gram-negative bacteria, new octacationic zinc phthalocyanines (ZnPcn+, n=4 or 8) offers the attractive prospect for improving both the water solubility and the localization of photoactivatable drugs in bacteria. We designed and synthesized hydrogel containing growth factors and PSs to improve the bactericidal activity of burn infection for topical application in clinics.

Tetrapyrrolic photosensitizers are currently highly attractive targets for the development of novel therapeutic agents not only for PDT, but also photoinactivation of microorganisms (PDI). Herein, we compare the physicochemical and photophysical properties of the series of structurally-related halogenated porphyrins in the homogeneous and heterogeneous systems in the context of their PDI applications. The mechanistic studies concerning ROS generation by these compounds are also highlighted. The results indicate that chemical modification of the macrocycle by tunable substituents as well as their impregnation on TiO2 surface provide an access to biological properties needed for efficient treatment without the development of resistance mechanisms.

In a clinical infection, microbial cells are present in distinct growth phases and it is important for any antimicrobial therapy to be equally effective in all different situations. This study evaluates molecular targets of Photodynamic Inactivation (PDI) mediate by methylene blue and a 660nm LED over Candida albicans at exponential growth phase in an attempt to identify the targets and therefore the potential for microbial recovery after PDI. Our results demonstrated that cells in exponential growing phase the internal targets as well nucleic acids are the main structures molecularly affected after exposure to PDI.

Cryo-electron tomography (cryo-ET) is an emerging technology that enables thin samples, including small intact prokaryotic cells, to be imaged in three dimensions in a near-native 'frozen-hydrated' state to a resolution sufficient to recognize very large macromolecular complexes in situ. This study used cryo-ET to evaluate the photodynamic effect on the viability and envelope architecture of a Gram-negative bacteria. E. coli minicells were submitted to photodynamic treatment and analyses using Cryo-ET. The imagens showed detachment of bacterial cell walls and mesosome-like structures. Even in sites where rupture was not observed, bacterial envelope was thicker, with relatively low density Cryo-electron tomography revealed that the effects of photodynamic therapy on Gram negative bacteria was based on damage to the outer membrane, cell wall and inner membrane and occurs in an energy-dependent manner.

Antimicrobial photodynamic inactivation (aPDI) is considered as an innovative alternative method to treat multidrug-resistant infections. we investigated the combinatorial antimicrobial therapy using a new water-soluble decacationic chlorin mediated aPDI with covalently-bonded vancomycin (VCMe-mChlPd-N10+) for killing methicillin-resistant Staphylococcus aureus (MRSA) upon illumination at 660 nm wavelength (40 mW/cm2) and 405nm wavelength (12 mW/cm2). VCMe-mChlPd-N10+ showed enhanced efficacy of aPDI of MRSA, compared with its analogous decacationic chlorin (ChlPd-N10+), non-cationic chlorin (ChlPd-EGn) and vancomycin alone. The study indicates VCMe-mChlPd-N10+ as a promising photosensitizer for aPDI of MRSA and provides a new approach of combinatorial antimicrobial therapy for localized multidrug-resistant infections.

Cutaneous Leishmaniasis affects the skin and mucous membranes. The treatment mostly effective and indicated for this disease have severe side effects evidencing the need for new treatment modality. Photodynamic Therapy (PDT) results in generation of reactive oxygen species, eradicating the parasite. The identification of the biochemical alterations that the therapy causes in the pathogen may help improve therapeutic efficiency. This study reports the use of FT-IR to investigate the chemical profiles of Leishmania major promastigotes in culture after PDT and compared with untreated pathogens. Results suggest increase in proteins, nuclear material, and decrease in lipids and carbohydrates.

Non melanoma skin cancer (NMSC) can be seen as a multifaceted problem and, based on this several problems, Photodynamic Therapy for small skin lesions may be one of the best solutions from an economic point of view. Using a strategy involving companies, national bank and medical partners, equipment, medication and protocols were tested in a multicenter study over Brazilian territory. With results collected over 5 years from a national program to implement PDT for non melanoma skin cancer, it was possible to reach a great economic evaluation of advances concerning the use of PDT for skin cancer.

Nitromedicine is a new medical specialty that focuses on developing diagnostic and the therapeutic protocols to enhance stem cell therapies and manage oxidative stress related diseases by modulating nitroredex sensitive epigenetics and biochemical pathways. The effect of ROS on oxidative stress related diseases is double edged sward . The production of ROS during normal signaling is essential for regeneration and healing effects , while excessive ROS production and consequent ROS accumulation lead to detrimental processes that cause degeneration and cellular injury. Oxidative stress related diseases occurs when antioxidant defenses cannot balance the production of ROS. The age-dependent Nitroredox imbalance is closely related to oxidative stress. ROS overproduction is involved in several oxidative stress related diseases such as degenerative diseases , autoimmune diseases , metabolic diseases , chronic inflammatory diseases , chronic infectious disease , cancer and pain conditions. To develop an effective therapeutic approaches against Oxidative stress related diseases , it is important to identify the mechanisms underlying the initial pathological events. Targeted antioxidant treatments could be a promising therapeutic approach. However, the complex nature of Oxidative stress related diseases and failure of monotherapies suggest that an antioxidant therapy should be accompanied by nitroredox multi-therapeutic interventions to enhance the restoration of the Nitroredox balance.

We intend to irradiate a laser of 664 nm through a laser catheter to the myocardium, shortly after the photosensitizer injection. The photosensitizer distribution would change drastically after the injection. We studied the dependence of the necessary time for myocardial cell necrosis on the photosensitizer contact time to find the optimal timing of laser irradiation n our proposed catheter ablation application. We found that the necessary time for cell necrosis decreases with the drug-light interval until around 15 min. We suggest that the effective timing for the photodynamic reaction with short drug-light interval exist because of the photosensitizer uptake process.

To investigate the use of photochemically triggered manipulation of cell signalling pathways, we exposed mouse osteoblast precursor cells and rat primary mesenchymal stromal cells to low-dose PDT. Our results demonstrate that low-dose PDT can promote osteoblast differentiation via the activation of activator protein-1 (AP-1). Although PDT has been used primarily as an anti-cancer therapy, the use of light as a photochemical “molecular switch” to promote differentiation should expand the utility of this method in basic research and clinical applications.

Chlorin-e6 (Ce6) is the one of the PS. We performed photodynamic diagnosis (PDD) using Ce6 in pancreatic cancer. We selected mouse tumor diameter about 5mm and Ce6 was injected in tail-vein. As a result, highly Ce6 was accumulated in tumor spot. Then shoot the light at the cancer place and we confirmed fluorescence. In result, intensity was decreased after PDT, we will study how to find pancreatic tumor region using laparoscopy and apply PDD and PDT. If this experiment was successfully completed, Ce6 can be applied for pancreatic cancer diagnosis and treatment simultaneously.

In our study, 808 nm near infrared light excited upconversion nanoparticles (UCNPs) were simultaneously loaded with the photosensitizer chlorin e6 (Ce6) and the glypican-3 (GPC3) antibody that isoverexpressed on hepatocellular carcinoma cells but not on normal liver tissue. The multitasking UCNPs@mSiO2-Ce6-GPC3 nanoparticles under 808 nm laser irradiation with enhanced depth of penetration would enable effective targeted PDT. We found the UCNPs@mSiO2-Ce6-GPC3 nanoparticles had good biocompatibility, low toxicity, excellent cell imaging in HepG2 cancer cells and high anti-tumor effect in vitro and in vivo.

The aim of this study is to investigate the tumor imaging capability of i.v. administered indocyanine green (ICG) in a rat colon carcinogenesis model. The ICG fluorescence was observed in the colon tumors 1 day after injection of ICG. ICG fluorescence in the tumor tissues was localized in the stromal cells at the vascular interstitial tissue of the luminal surface. These results suggest that fluorescence imaging following the administration of i.v. ICG can detect the colon tumors. The tumor tissue preference of ICG in the current model can be attributable to the stromal cellular uptake and retention of ICG.

While many have contributed to the field of ‘photodynamic therapy’, Thomas J. Dougherty played the central role. Without his efforts, it seems unlikely that PDT would have advanced much beyond being a medical curiosity. This summary discusses both his many contributions to the field and the wide range of discoveries made possible by his pioneering studies that brought PDT to the attention of the funding agencies.

Though my area of expertise (synthetic organic chemistry) is different from those of Tom Dougherty, we nonetheless managed to actively collaborate on several PDT projects through the intermediacy of Professor Ravi Pandey. This brief talk will concentrate on those projects that we published jointly, and on my memories of Tom, a great scientist, and an even greater friend.

The effect of PDT on the host immune response and the role the host immune response plays in PDT efficacy against cancer has been the subject of intensive research for the past 2 plus decades. During that time we have learned that 1) the patient’s immune health can affect the efficacy of PDT; 2) PDT can both enhance and suppression immunity; 3) treatment regimens can be devised to enhance anti-tumor immunity; and 4) PDT has the potential to be an effective immunotherapy. This presentation will focus on how these findings came about and the role Dr. Dougherty played in the discoveries.

Since 1978 the initial research on PDTand PDD using canine lung cancer model and then clinical research of early stage cancers of lung, esophagus, stomach and cervix was performed with hematoporphyrin derivative and argon dye laser. A lot of meetings on PRT which was called initially, mainly in USA, Late in 1980's IPA was founded during the International Meeting of Clinical Application of PRT in Tokyo. After the extended clinical trials of PDT on early stage cancers by the support of Government in 1980' Japanese Government approved PDT with Photofrin and argon dye laser or excimer dye laser for clinical use in 1992. A New photosensitizer of Laserphyrin ( Chrolin e6) wad developed after the preclinical investigation by Prof. David Kessel. This photosensitizer showed less skin photosensitibity than previous Photofrin.

Maximizing extent of resection for brain tumors has been shown to improve patient outcomes. Fluorescence guided surgery (FGS) is a safe and effective adjunct improving extent of resection. Nevertheless, state-of-the-art FGS uses qualitative assessments of the fluorescence which suffers from substantial sensitivity limitations in HGG and more significantly in LGG. Quantitative fluorescence assessments, which correct for the distorting effects of tissues optical properties, have demonstrated improved diagnostic accuracies across a broad range of tumors including high and low grade gliomas, meningiomas, and metastases. Here we discuss the advantages and disadvantages of qualitative and quantitative fluorescence in FGS.

Standard chemoradiation often enriches drug-resistant tumor cell populations that can lead to recurrent and treatment-refractory disease. For instance, preclinical models of several cancers suggest that the cancer stem cell subpopulation becomes enriched and re-populates the tumor milieu following conventional therapies. Here, we show evidence that photodynamic therapy (PDT) is effective against several patient-derived glioblastoma stem cell cultures. Moreover, sub-lethal PDT results in re-sensitization of cancer cell phenotypes with induced drug-resistance to chemotherapy. This talk will also introduce some of our efforts to establish a new program aimed at developing personalized and targeted PDT to overcome drug-resistance.

The efficacy of Rutherrin based on the Photosensitizer TLD1433 was evaluated in the RG2 preclinical glioma model in vitro and in vivo. The in vitro LD50 as a function of photons absorbed by the photosensitizer was lower than for ALA-induced PpIX using previously published protocols. In vivo Rutherrin demonstrated a higher specific uptake ratio, longer survival times, lower edema and a higher infiltration of CD8+T immune-regulating cells and the in vivo threshold (number of photons absorbed to cause cell death) is two orders of magnitude lower compared to ALA-induced PpIX treated animals.

Glioblastoma is a malignant brain tumor with a median overall survival of approximately 15 months with the current standard of care and for which adjuvant therapies are highly expected. 5-ALA PDT have been reported with promising results to treat glioblastoma. We present here a clinical trial to evaluate 5-ALA PDT delivered intraoperatively to treat newly diagnosed GBM. Ten patients have been enrolled. Currently, therapy has been delivered without significant toxicity or adverse event and are fulfilling the primary endpoint of this feasibility study. Secondary endpoints still being under investigation are progression-free survival, overall survival and patients' quality of life.

We studied the correlation between preoperative positron emission tomography with 11C-Methionine (Met-PET) and intraoperative 5-aminolevulinic acid (5-ALA) fluorescence in glioma surgery. 16 cases with astrocytic tumors were analyzed. Fluorescence intensity was categorized into four groups. The regional uptake of Met-PET was significantly high in the strongest fluorescence group in newly-diagnosed cases (p=0.017). This group showed significant difference when compared with other groups together, in all cases and newly-diagnosed cases (p=0.01 and p=0.004, respectively). In our study with objective spectroscopic measurement of fluorescence, strong 5-ALA fluorescence reflected high Met-PET uptake. This could indicate the importance of bright fluorescence by 5-ALA.

Here we demonstrate that photodynamic effects (PD) opens the blood-brain barriers (BBB) that is new important informative platform for development of perspective strategies for brain drug delivery including liposomes as a good candidate for drug delivery system.

This talk will introduce spatiotemporal synchronization of photodynamic therapy (PDT) combined with molecular targeted therapies. We newly developed photoactivatable multi-inhibitor nanoliposomes (PMILs) for photodynamic tumor cell and microvessel damage in concert with photo-initiation of tumor-confined, multikinase inhibitor release. Preclinical data in mouse models of pancreatic cancer demonstrates that this concept facilitates suppression of the VEGF and MET signaling pathways—both critical to cancer progression, metastasis and treatment escape. Remarkably, a single PMIL treatment using low doses of a multikinase inhibitor (less than 1/1,000th of the standard cumulative dose) achieves sustained tumor reduction and suppresses metastatic escape. The PMIL concept is amenable to a number of molecular inhibitors and offers new prospects for precise and efficient use of potent but toxic multimolecular targeted therapies, reducing systemic drug exposure and associated toxicities.

Considerable attention has been given to improving the photoactivity and biocompatibility of hydrophobic photosensitizing drugs. It is increasingly clear that photosensitizing biomolecules, based on chemical conjugation or association of photosensitizers with biomolecules, strongly influence the performance of a given photosensitizer in biological environments. However, the numerous studies that have revealed PSBMs are not readily comparable as they cover a wide range of macromolecules, evaluated across a range of experimental conditions. Here, we prepared and characterized a series of well-defined PSBMs and pure drug crystal based on a clinically used photosensitizer—benzoporphyrin derivative (BPD). Our results illuminate the variable trafficking and end effects of clinically relevant PSBMs and BPD nanocrystals, and demonstrate that biologically-informed combinations to target multiple subcellular organelles may lead to enhanced therapeutic effects in gliomas.

In this study we tested the ability of Indocyanine Green (ICG, a photosensitizer) loaded plasmonic gold nanorods (AuNP) assisted combined photothermal and photodynamic therapy for laser induced choroidal neovascularization (CNV) in pigmented mice. Near infrared laser (810 nm) was used for the therapy. In-vivo ocular imaging (SD-OCT and cSLO) revealed increased CNV healing and decreased leakage with treatment. ROS induced DNA damage was assessed by immunohistochemistry with 8-Hydroxyguanosine antibody using confocal microscopy imaging. These results suggest that ICG-AuNP allows localized and targeted photodynamic/photothermal therapy while minimizing laser-induced damage to healthy tissues.

The present study is aimed at the development of drug-in-cyclodextrin-in-liposome nanoparticles by coupling two independent delivery systems: cyclodextrin/temoporfin inclusion complexes and liposomal vesicles to improve the transport of temoporfin to target tissue. Liposomes offer an excellent opportunity to achieve selective drug targeting what is expected to prevent local irritation and reduce drug toxicity. Cyclodextrins have been utilized as independent carriers for the improvement of temoporfin pharmaceutical properties such as solubility, stability, and bioavailability. Therefore, it was assumed that encapsulation of cyclodextrin-complexed drugs into liposomes may increase drug loading capacity and restrain the dissociation of drug-cyclodextrin complexes and prolong its systemic circulation.

The molecular properties that contribute to the potency and to the efficacy of photosensitizers are discussed with special emphasis on porphyrins, chlorins and bacteriochlorins. Amphiphilicity, size, molar absorption coefficients, photostability, lifetimes, triplet and singlet oxygen quantum yields, cell uptake and subcellular localization are related to potency and efficacy. Examples are given to illustrate the impact of such factors, namely using structure-activity relationships and isomers. The concentration of reactive oxygen species generated, their subcellular localization and tissue distribution are critical factors for potency and efficacy.

Transition metal complexes are of interest for their use as photosensitizers (PSs) in light-based applications, including photodynamic therapy (PDT). Certain coordination complexes of ruthenium (Ru) yield potent PDT effects in a variety of in vitro and in vivo cancer models. One example is our own TLD1433, which is the first Ru PS to enter (and successfully complete) a human clinical trial for treating cancer with PDT. The design and development of transition metal complexes for PDT will be discussed.

Translocator protein (TSPO) is an attractive target for cancer treatment due to its over-expression in highly aggressive tumors. We report here new TSPO-targeted agents for photodynamic therapy (PDT). We synthesized three TSPO ligands, optimized their binding affinities and conjugated them to a photosensitizer, IR700DX. The tetrahydrocarbazole-based TSPO-PDT agent showed desirable nanomolar affinity to TSPO and significant PDT effects on aggressive cancer cells and tumors.

The possibility of fine modulation of the photophysical signatures and singlet oxygen production of BODIPY chromophores by a precise molecular design has been demonstrated. This dye can be postulated as a good PDT agent, since it: (1) synthetic accessibility, (2) robustness to endure relatively-high density photo-excitation, (3) efficient triplet-state population to enable generation of 1O2, (4) absence of dark toxicity, (5) ability to be immobilized on nanoparticles and (6) high biocompatibility to promote specific localization into cell. Moreover, brominated BODIPYs, biscyclometalated Ir(III)-BODIPY complexes or, particularly, orthogonal BODIPYs dimers (without using heavy atoms) could be used as biomaterials for theragnosis due to their high-enough fluorescence making bioimaging possible together with their ability to generate singlet oxygen cytotoxic species to efficiently kill cancer cells.

We investigated the efficacy of Ruthenium and Osmium-based coordination complexes as potential photosensitizers (PS’s) for photodynamic therapy (PDT). These compounds act as an alternative to porphyrin-based PS’s, and are capable of activation over a broad range of wavelengths. Compounds were tested at 532 nm and 630 nm wavelengths in-vitro, using metastatic murine melanoma (B16F10), murine squamous cell carcinoma (SCCVII), and human adenocarcinoma (A549) cell lines. Results revealed effective PDT-mediated cytotoxicity in all three cell lines, with minimal dark toxicity observed. These novel PS’s are anticipated to be effective against multiple cancers.

The intention of this study was to improve the photosynthetic drug delivery of ZnPcS4 in lung cancer cells, by enhancing its chemical structure. ZnPcS4 was successfully conjugated to pegylated gold nanoparticles, as well as bound to specific active tumour-associated antibody-antigens (Cetuximab) to aid specific targeted PS delivery. Within in vitro lung cancer (A549) cells, this molecular drug conjugate proved to have enhanced cellular uptake. Moreover, after conducting in vitro PDT experiments, a significant amount of cell death and cytotoxicity was noted, suggesting that this molecular drug conjugation combination proved to enhance the PDT treatment capabilities for lung cancer

This talk will focus on the properties of two classes of photosensitizers, namely pyridone-substituted porphyrins and BODIPY dyads, both of which generate singlet oxygen and have aromatic units capable of [4+2] cycloaddition reactions with singlet oxygen. A discussion on their applicability to phototherapy will be accompanied by in vitro evaluation.

Five-aminolevulinic acid (5-ALA) derived tumor porphyrins are being explored for a variety of tumors of the CNS for fluorescence-guided surgery (FGS) and more recently, for photodynamic therapy. Nevertheless, the main application remains FGS for malignant gliomas. This paper will briefly review the regulatory history of 5-ALA FGS, the state-of-the art applications for high grade glioma surgery and possible future directions in intra-operative porphyrin fluorescence detection. In this regard it will also discuss some of the possible pitfalls and hurdles potentially involved in establishing such technologies in order to ensure ongoing safe and effective use of fo 5-ALA FGS.

This Phase II RCT evaluated the performance of a handheld imaging device, PRODIGI, combined with the pro-drug 5-aminolevulinic acid (5-ALA) HCl for intraoperative visualization of invasive breast carcinomas. Patients received 0, 15 or 30 mg/kg 5-ALA HCl (n = 15/group). 5-ALA improved tumor-to-normal contrast and visualization of clinically occult carcinoma. Positive predictive value for detecting tumor inside and outside the demarcated tumor border was 100.0% and 55.6%, respectively (15 mg/kg dose group) and 100.0% and 50.0%, respectively (30 mg/kg dose group). Technical feasibility and clinical integration were confirmed. This is the first report of intraoperative visualization of 5-ALA-induced fluorescence in breast cancer using a fully handheld imaging device.

Clinical investigations of 5-ALA based iPDT for malignant glioma showed promising longterm progression free and overall survivals. High light fluences are applied with our approach for iPDT, aiming for a deep treatment penetration into the infiltration zone, while preserving functional brain thanks to the highly selective accumulation of photosensitizer and fast photobleaching. Study of PDT induced immune response and individualization of treatment parameters may lead to further improvement of clinical outcomes.

We examined the usefulness of 5-ALA-FD (5-ALA fluorescence guided detection) for detection of various cancers by reviewing our collaboration studies conducted in Japan. 5-ALA was orally administrated at 20mg/kg or 1g per patient 2-4 hours before surgery or diagnosis. We focused on effectiveness of 5-ALA-PD to detect the followings: 1) carcinoma in situ of non-muscle-invasive bladder cancer, 2) invisible peritoneal metastases of gastric cancer, 3) peritoneal nodules of pancreatic cancer, 4) preoperative diagnosis of lung cancer. Those studies indicate that 5-ALA-FD is useful for distinction of small tumor lesions and contributes to selection of an appropriate therapy without non-curative resection

Aminolevulinic Acid and its derivatives are the most common photosensitizer’s precursors allowing the Protoporphyrin IX accumulation in topical Photodynamic Therapy. We tested dissolving MNs using 5% ALA concentration applied in three healthy volunteers; OCT images and the superficial PpIX formation of the dissolving MNs were compared with the 20% MAL cream. The study showed even with a lower concentration, the MNs were able to produce a similar amount of PpIX compared to the cream in the same incubation time. The results were encouraging to perform an animal tumor model to support the understanding of the PDT.

The purpose of this research project was to demonstrate the the efficacy of photodynamic therapy (PDT) as an alternative to antibiotic therapy for the treatment of infected chronic LUs. PDT using irradiation at fluency 80 J / cm2 and 20% 5-aminolevulinic acid topically applied for 4 hrs, was performed during 10 days of hospitalization in 10 patients both sexes aged 40-85, with chronic leg ulcers, who had not responded well to conventional treatment. This was a randomized, placebo-controlled trial (local Octenidine dihydrochloride). The numerical scale of pain was used. Treatments were carried out at 3-week intervals. During 8 months follow up, complete remission (CR) was obtained in 4 patients (40%), partial response (>50% reduction in ulcer diameter), in 3 patients (30%), no response in 1 patients (10%). In two patients (20%) was observed deterioration of the local condition, with swelling, erythema and inflammation.

A total of 62 female patients who underwent ALA-PDT at the Gynecology Department of Henan Province People's Hospital in 2018 were retrospectively assessed. Lesions included 24 cases of LSIL, 13 cases of HSIL, 12 cases of VAIN, 2 cases of VIN and 11 cases of VL. Three sessions of ALA-PDT were carried out at intervals of ten days. After six month follow-up, The effective rate of LSIL、HSIL、VAIN、VIN and VL were 87.5%、92.3%、91.7%、100% and 100% . ALA-PDT is an effective treatment for female genital precancerous lesions.

Simultaneous metabolic and oxygen imaging is a promising idea to follow up therapy response, disease development and to determine prognostic factors. A common property during tumor development is altered energy metabolism, which could lead to a switch between oxidative phosphorylation(OXPHOS) and glycolysis. The impact of this switch for theranostic applications is significant. FLIM of metabolic coenzymes, as NADH and FAD, is now widely accepted to be the most reliable method to determine cell metabolism. During photodynamic therapy (PDT) also oxygen consumption has to be taken into account to understand treatment responses. Interestingly, the phosphorescence lifetime of phosphorescent photosensitizers (PSs) is able to indicate local oxygen changes. Within this work, correlated imaging of PS phosphorescence and fluorescence lifetime parameters was implemented and will be discussed for new theranostic applications. The technique for simultaneous FLIM/PLIM is based on time correlated single photon counting (TCSPC). For this a laser scanning microscope (LSM 710, Zeiss) was equipped with a two channel TCSPC system (Becker and Hickl, Berlin) and a fs pulsed Ti:Sa Laser (MaiTai DeepSee, Spectra Physics) for short pulsed two photon excitation. The photosensitizer TLD1433 (Theralase Inc. 1945 Queen St E, Toronto, ON M4L 1H7), which is based on a ruthenium (II) coordination complex, was used for PLIM. TLD1433 possess a variety of different triplet states, which enables complex photochemistry and redox reactions (see Jablonski diagram). TLD1433 can be used as a phosphor to follow up local oxygen concentration and consumption during PDT. Results obtained in human bladder carcinoma cells will be discussed.

TDL1433 mediated PDT for NMIBC for patients who failed standard BCG immunotherapy was shown to be safe and well tolerated at the therapeutic dose. Contributing to the safety of the treatment is instilling the PS to minimize pharmacokinetic influences on the drug dose, and monitoring the irradiance on the bladder wall to minimize optical effects when illuminating a cavity with unknown optical properties. Light source limitations resulted in long irradiation times, and higher total power fiber optical delivery must be attained for clinical translation. The exploratory efficacy data is encouraging.

Surgical management of peritoneal metastases raises the problem of the theoretical involvement of the entire peritoneum. Intraperitoneal photodynamic therapy (PDT) has been limited by the lack of specificity of photosensitizers and difficulties to bring light into the cavity. Recent data may give a new breath for PDT. Indeed, targeted molecules are being developed, and optimization of illumination systems allows the development of PDT by minimally-invasive approaches. Expected abscopal effect of PDT could enhance a systemic immune response. The objective of this review is to present the most recent data and possible approaches for the treatment of peritoneal carcinomatosis by PDT.

A total of 50 patients with histologically confirmed cervical LSIL were treated with ALA-PDT at the Gynecology department of Ren Ji Hospital, School of Medicine, Shanghai JiaoTong University. Three sessions of ALA-PDT were carried out at intervals of seven days. The efficacy was evaluated by TCT, HPV typing and colposcopy directed biopsy. Three months follow-up demonstrated that the overall response rate for cervical LSIL was 74% (37 / 50), and the total virus remission rates was 66% ( 33 / 50). In addition, age and HPV type didn’t affect HPV remission rate. ALA-PDT is an effective treatment for cervical LSIL.

Ovarian cancer is the most lethal gynecological cancer in women. In this study, we evaluated both in vitro and in vivo the effect of a new photosensitizer (PS2) on ovarian peritoneal ovarian cancer. We showed that, in vitro and upon 1h of illumination, the PS2-PDT was capable of inducing ovarian cancer cell death and activate the mitochondrial metabolism of some immune cells. Furthermore we developed a mouse model of ovarian cancer stably expressing luciferase in OVCAR-3 cell-line allowing us to follow tumor development. The PS2 was well tolerated by mice and our preliminary results showed a tumor regression upon illumination.

This study aims to compare PDT and administration of 70% trichloroacetic acid (TAA) in condylomas. The protocol involved application of 20% methyl aminulevulinate cream and the illumination was performed with a total dose of 150J/cm², showing the need to develop new devices. 22 patients have been evaluated, totaling 15 patients treated with TAA and complete response of 33% and 8 with PDT (complete response of 75%). It is important to highlight that 2 patients showed recurrence for TAA and 4 patients discontinued the treatment with TAA, while, for PDT, there were no cases both recurrence and withdrawal.

Summary High grade cervical intraepithelial neoplasia (HCIN) is considered a precursor lesion of cervical cancer that can limit the young women reproductive, mainly if it progresses to invasive disease. Attempting to reduce the uterine cervix loss, which is considered standard treatment to treat HCIN, we suggest Photodynamic therapy. This treatment reduces viral load, return Pap smear to normal standards, preserves the cervix and enabling future pregnancies. The possibility of application in developing countries can be considered an innovation, in addition, reducing treatment costs.

The beginnings of photodynamic therapy date from 1900 but mechanisms relating to photokilling were unknown until 1991 when Oleinick’s group first described PDT-induced apoptosis. Routes to apoptosis have been determined along with the role of sub-cellular photodamage as a factor in initiation of cell death. We have recently reported on the role of paraptosis, another death mode that can be operative in cells with an impaired apoptotic program. It appears that the sensitivity of malignant cell types to lethal photodamage can be magnified by careful selecting of sub-cellular targets. In this report, recent progress relating to potential routes to improved PDT responses is described.

Photochemical internalization (PCI) is a drug delivery modality where PDT with an amphiphilic photosensitizer, is combined with a molecular drug which accumulate in endocytic vesicles. Light exposure destabilize the endo/lysosomal membrane so that the macromolecule is released into the cytosol where it can exert its therapeutic effect. PDT-induced induction of immunogenic cell death is highly dependent on the chemical composition of the photosensitizer and its intracellular localization. Little is, however, known on immunogenic cell death following PDT with PCI relevant photosensitizers such as TPCS2a. Also the macromolecular drug used in PCI may impact on the immunogenic treatment response. In the present study we elucidated mechanisms of immunogenic cell death following PCI of a VEGFR targeting toxin, VEGF121/rGel.

Photodynamic therapy (PDT) represents a possible alternative to ionizing radiation and chemotherapy for breast cancer. During PDT, 5-aminolevulinate (ALA) is converted into protoporphyrin IX (PpIX) within mitochondria, then activated by light. Systemic 5-fluorouracil (5FU) prior to ALA increases PpIX levels and improves efficacy, but toxicity remains a concern. Here, we employed Capecitabine (CPBN), a 5FU precursor selectively metabolized to 5FU within cancer cells, as a neoadjuvant. In a murine 4T1 breast cancer model, a CPBN/PDT combination enhanced tumor cell differentiation, PpIX levels, and tumor cell death. It also reduced metastatic spread. This combination approach offers significant potential for clinical translation.

A red absorbing, water soluble and mitochondria targeting BODIPY based photosensitizer was realized and its potential as photodynamic therapy agent was evaluated. The photosensitizer not only effectively functioned under hypoxic conditions but also showed highly selective photocytotoxicity towards cancer cells. To the best of our knowledge this marks the first example of a mitochondria targeted BODIPY photosensitizer that functions under hypoxia.

Photodynamic therapy (PDT) and diagnosis (PDD) using 5-aminolevulinic acid (ALA) to drive the production of an intracellular photosensitizer, protoporphyrin IX (PpIX), are in common clinical use. Dormant cancer cells, one of factors of recurrent metastatic cancer, whose physiological functions pause or become quiescent are relatively insensitive to most chemotherapeutic drugs and radiation. We demonstrated PC-3, prostate cancer, exhibited dormancy in a cell density-dependent manner not only in 2D culture but in 3D culture. Dormant cancer cells accumulated high PpIX levels and were sensitive to ALA-PDT. Furthermore, PpIX accumulation and ALA-PDT cytotoxicity were enhanced by G0/G1-phase arrestors in non-dormant cancer cells.

For four decades, our research has focused on one challenge: improving the delivery and efficacy of anti-cancer therapies. Working on the hypothesis that the abnormal tumor microenvironment fuels tumor progression and treatment resistance, we developed an array of novel imaging technologies and animal models as well as mathematical models to unravel the complex biology of tumors. Using these tools, we demonstrated that the blood and lymphatic vasculature, fibroblasts, immune cells and the extracellular matrix associated with tumors are abnormal, which together create a hostile biochemical and physical tumor microenvironment (e.g., hypoxia, high interstitial fluid pressure, high solid stress). Our work also revealed how these abnormalities fuel tumor progression and metastasis, while preventing treatments from reaching and attacking tumor cells. We next hypothesized that if we could reengineer the tumor microenvironment, we should be able to improve the treatment outcome. Indeed, we demonstrated that judicious use of antiangiogenic agents—originally designed to starve tumors—could transiently “normalize” the tumor vasculature, alleviate hypoxia, increase delivery of drugs and anti-tumor immune cells, and improve the outcome of radiation, chemotherapy and immunotherapy in a number of animal models. Moreover, our trials of antiangiogenics in brain and breast cancer patients supported this concept. They revealed that the patients whose tumor blood perfusion/oxygenation increased in response to anti-VEGF therapies survived longer than those whose blood perfusion/oxygenation did not increase. The normalization hypothesis also explained how anti-VEGF agents could improve vision in patients with wet age-related macular degeneration, and opened doors to treating other non-malignant diseases harboring abnormal vasculature that afflict more than 500 million people worldwide [e.g., neurofibromatosis-2 (NF2), which can lead to deafness; tuberculosis; plaque rupture]. Our clinical finding led to the approval of bevacizumab for NF2 patients in UK in 2014. In parallel, by imaging collagen and measuring diffusion and perfusion in tumors in vivo, we discovered that the tumor cells and the extracellular matrix compress blood vessels and impede drug delivery in desmoplastic tumors (e.g., pancreatic cancer, triple negative breast cancers). We subsequently discovered that angiotensin blockers – widely prescribed to control hypertension – are capable of “normalizing” the extracellular matrix, opening compressed tumor vessels, and improving the delivery and efficacy of molecular and nano-therapeutics. This finding offers new hope for improving treatment of highly fibrotic tumors and led to a successful phase II clinical trial at MGH on losartan and chemo-radiation therapy in pancreatic ductal adenocarcinomas (PDAC) (NCT01821729). This trial demonstrated that the addition of losartan to the standard of care led to an unprecedented R0 resection rate of 61% in locally advanced PDAC and significantly improved survival of these PDAC patients. This finding has led to a multi-institutional randomized trial. In my presentation I’ll also discuss how these two broad strategies – “vascular normalization” and “matrix normalization” – can improve the delivery and efficacy of various cancer therapies, including immunotherapy.

To understand the origin of life on Earth, and to evaluate the potential for life on exoplanets, we must understand the pathways that lead from chemistry to biology. Recent experiments suggest that a chemically rich environment that provides the building blocks of membranes, nucleic acids and peptides, along with sources of chemical energy, could result in the emergence of replicating, evolving cells. Diverse photochemical processes are thought to have been important for the origin of life, from photochemical steps in prebiotic synthetic pathways, to the generation of useful forms of chemical energy, to the selection of the canonical nucleotides based on their photostability. I will discuss the many ways in which UV light may have influenced the origin of life.

The clinical proof of concept that modulation of the immune system could lead to meaningful therapeutic benefit in cancer treatment was propelled into the public eye with the approval of anti-CTLA-4 immunotherapy ipilimumab in 2011 for unresectable or metastatic melanoma. A so-called “checkpoint inhibitor,” CTLA-4 is expressed on the surface of T cells and can be exploited by tumors to evade an anti-tumor immune response by turning off T cell proliferation and activation. Blockade of another checkpoint inhibitor, PD-1, or its ligand expressed on tumors called PD-L1, demonstrated higher response rates across multiple tumor indications with an improved safety profile. Interestingly, co-blockade of CTLA-4 and PD-1 has led to enhanced clinical benefit – with both greater response rates and durability – which has led to a new wave in combination immunotherapy, targeting next-generation checkpoint inhibitors as well as novel nodes of the suppressive tumor microenvironment. Next-generation checkpoint inhibitors including TIM-3 and LAG-3 have broad expression profiles, and preclinical research reveals novel and critical mechanisms of action for these pathways. Translational data from clinical trials also informs understanding of novel mechanisms. Further next generation targets in development, including combinations of immunotherapy with standard of care chemotherapy and targeted therapy, and novel targets of suppressive molecules/pathways in the tumor microenvironment, will expand the repertoire of key immuno-modulatory agents to harness the anti-tumor immune response.

The use of endoscopic optical coherence tomography (OCT) in pulmonary medicine has shown significant promise for aiding in the detection and diagnosis of airway pathology, in monitoring disease progression over time, and in assessing dynamic microstructure and function in vivo. Recent technical advances including enhanced resolution and contrast together with novel sophisticated catheter designs to improve usability are key contributing factors to the increased clinical utility of OCT in pulmonology. We will discuss some of these recent advancements and present results from clinical studies highlighting the potential impact for patient care.

Introduction/Aim: Photodynanic therapy (PDT), is a treatment modality for many cancers, and uses a tumor-specific photosensitizer and laser irradiation. Recently, we have developed a laser probe In this study, we aimed to develop a new endobronchial treatment for peripheral cancer using PDT, and we evaluated the feasibility of PDT for peripheral lung cancer. Methods: This phase I study enrolled 7 patients with peripheral lung cancers (primary tumor< 20 mm, stage IA), which were definitively diagnosed by bronchoscopic modalities such as EBUS-GS and brocnhoscopic navigation system. Results: We performed PDT for 3 patients with c-stage IA peripheral lung cancer, using a laser dose (120mW, 50J/cm2), and confirmed the feasibility of the dose. We escalated the laser dose and performed 4 patients using a laser dose (120mW, 100J/cm2). Seven patients met our criteria, and 5 cases were adenocarcinoma and 2 case squamous cell carcinoma. Two weeks and 3 months after NPe6-PDT, complication

Abstract

5-ALA is approved in the diagnosis of brain tumor and bladder tumor by the Ministry of Health, Labour and Welfare. 5-ALA is a natural amino acid contained in a living body and is a precursor of hemoglobin. When absorbed into the body by oral ingestion, porphyrin synthesis by heme synthesis is specifically performed in a tumor-specific manner, and accumulation of fluorescent protoporphyrin IX (pp IX) is observed. However, pp IX is metabolized and disappeared in normal tissues. When tumor tissue is irradiated with excitation light, at the site where pp IX accumulates, it exhibits red fluorescence. We applied 5-ALA for photodynamic diagnostic of thoracic malignancies.

The iPDT multimodal treatment for MPM is applicable and manageable in a European expert center, involving local skills and dedicated teams. The safety profile of the iPDT in Lille center was favorable, as validated by an external board. Median overall survival was promising (≈28 months), similar to previous US results. Our center is expected to join soon a large phase II randomized, multicentric US trial assessing MPM multimodal treatment (P/D, chemotherapy) ±iPDT (NCT02153229; UPENN, USA).

The report of patients with lung cancer who could receive PDT combined with other modalities like surgery and chemotherapy is relatively rare. Combination of PDT and surgery is useful for multiple lung cancers or reduction of resection line of superficial invasion. Also, PDT combined with chemotherapy for advanced lung cancer with central airway stenosis seems to be usefull for local control and improvement of patient’s QOL. In MPLCs, CRs were achieved by surgery combined with PDT in all patients. After induction PDT, reduction surgery could be successfully performed in 82%. In PDT combined with chemotherapy, all patients improved symptoms and QOL after treatment. PDT combined with other modalities may be a promising strategy in lung cancer treatment.

Operation, chemotherapy and radiotherapy are typical treatments of lung cancer today. Curative tumor resections are still the mainstream treatment for early-stage small peripheral lung cancer. However, non-surgical treatments such as trans-bronchoscopy ablation, including bronchoscopy microwave ablation and bronchoscopy PDT, are also potential options. We proposed a novel Photodynamic Therapy technique using Lipiodol as light scattering agent for the peripheral lung tumor. Lipoidol (high refractive index 1.49) was injected into bronchus and light can delivered in the bronchus similar as optic fiber. We also found that dissolving Indole-3-acetic acid (IAA) in Lipiodol could create a new drug, which can be used as light scattering agent and oxidant as a novel photodynamic therapy method.

Photochemical internalization (PCI) is a technology enhancing intracellular drug delivery by light-induced translocation of endocytosed therapeutics into the cytosol. PCI of BLM did not induce a curative effect in athymic mice while more than 90% of the thymic mice were cured. Cured thymic mice, re-challenged with CT26.CL25 tumor cells on the contralateral leg, rejected 57-100% of the tumors inoculated up to 2 months after PCI. T-cells from the spleen of PCI-treated mice were found to inhibit the growth of CT26.CL25 cells in naïve thymic mice showing that treatment of CT26.CL25 tumors in thymic mice by PCI induces systemic anti-tumor immunity.

Enhancement of anti-tumor immunity by PDT is dependent upon activation of T cells, which is mediated by the extent of acute inflammation occurring within 4-8 hours post-PDT. The acute inflammation is characterized by neutrophil migration into the treated tumor and tumor draining lymph node. The increase in T cell activation is accompanied by increased expression of IFN-γ. Enhanced anti-tumor immunity correlates with increased expression of immune checkpoint inhibitory molecules on tumor and immune cells. Addition of immune checkpoint inhibitors improves both the efficacy of PDT and the functional status of anti-tumor T cells following PDT.

Photodynamic therapy (PDT) is an effective anticancer procedure that relies on tumor localization of a photosensitizer followed by light activation to generate cytotoxic reactive oxygen species (e.g., 1O2). We recently reported nanoscale metal-organic frameworks (nMOFs) as exceptionally effective photosensitizers for PDT of cancers. These nMOFs efficiently generate 1O2 owing to site isolation of photosensitizing ligands and facile 1O2 diffusion through porous nMOFs. Consequently, nMOFs displayed greatly enhanced PDT efficacy both in vitro and in vivo, leading to complete local tumor eradication in the mouse models. We further combined nMOF-mediated PDT and an immunotherapy agent to elicit systemic antitumor immunity. We elucidated the underlying immunological mechanisms for the combination therapy. We believe that nMOF-enabled PDT has the potential to significantly enhance checkpoint blockade cancer immunotherapy.

Cytokines are potent immunomodulatory agents which play important roles in countering the highly immunosuppressive tumor microenvironment. A major hurdle in systemic cytokine therapies is achieving effective therapeutic levels within the affected tissue with minimal systemic toxicity. While the pleotropic cytokine IL12 has been investigated with encouraging results in animal studies, clinical progress is hampered by low tumor uptake and high rate of adverse effects. Herein we propose a photo-triggered liposomal nanocarrier for IL-12 which provides spatio-temporal control on immunotherapy delivery to solid tumors by leveraging the enhanced permeation and retention effect for enhanced accumulation, followed by image guided light triggering to obtain tumor specific bolus release of IL-12. Liposome based cytokine delivery offers opportunities for synergistic combination treatment with other chemo and phototherapy agents as well.

Targeted photo-immunotherapy (PIT) may reduce the adverse effects of non-targeted PDT. Cetuximab-IRDye700DX showed significant tumor to normal ratio in the skin-fold chamber model using intra-vital microscopy. Treatment lead to vascular responses in tumor and normal tissue. Subcutaneous OSC-19 tumors were transdermally illuminated with 100 J.cm-2 at 20, 50 and 150 mW.cm-2. All animals treated with 20 mW.cm-2 showed no tumor up to 90 days post treatment (n=4) compared to 1 of 4 in the 50 and 150 mW.cm-2 groups. The remaining tumors reached 200% after 17.9±5.2 and 19.5±7.4 days. The effect of targeted-PIT is strongly dependent on fluence rate.

ALA-PDT is an effective treatment for actinic keratosis (AK). However, conventional PDT causes pain during illumination. In a relatively new regimen called metronomic PDT (mPDT) or painless PDT, the illumination is delivered concurrently with ALA application. We treated murine AK lesions with either mPDT or PDT followed by analyses of lesion clearance and mechanistic events. PDT and mPDT led to similar lesion clearance, decreased apoptosis, autophagy, and an immune response characterized by infiltration of neutrophils and macrophages. Preliminary data suggest that mPDT is just as effective as conventional PDT for treatment of AK, but the mechanisms may be quite different.

Photoimmunotherapy combines the phototoxicity of photosensitizer and targeting capability of antibody to improve potency and reduce the adverse effect of photodynamic therapy (PDT). This paper describes the conjugation of IRDye 700DX (IR700), a water-soluble photosensitizer, with anti-Cadherin-17 (CDH17) humanized monoclonal antibody (ARB102) to achieve targeted PDT in gastrointestinal (GI) cancers. CDH17 is a cancer antigen overexpressed in GI cancers with restricted normal tissue expression. ARB102-IR700 can target CDH17+ cancer cells and induce cytotoxicity at high potency and specificity upon light irradiation. Selective killing of CDH17+ cancer cells via ARB102-IR700 PDT and the underlying cell death mechanism are under investigation.

Photodynamic therapy (PDT) is an excellent treatment modality for superficial skin cancers. However, thick lesions may not respond. Here we describe new approaches to improve PDT treatment efficacy by manipulating the biology of tumor cells, specifically through the addition of 5-fluorouracil, methotrexate, or Vitamin D as combination agents (neoadjuvants). Another limiting aspect of PDT is severe pain during illumination; we report on a clinical trial in which PDT parameters were customized to greatly reduce pain. Finally, we describe a new clinical trial that was recently initiated to investigate the relationship between Vitamin D levels and PDT efficacy for BCC.

The most interesting aspect in 5-aminolevulinic acid (ALA)-mediated photodynamic therapy (PDT) on cutaneous squamous cell carcinoma (SCC) is its potential in inducing antitumor immune responses. It was found that ALA-PDT not only kills tumor cells directly but also rapidly recruits and activates immune cells favoring the development of antitumor adaptive immunity. Our study showed that ALA-PDT enhanced the expression of calreticulin (CRT), heat shock proteins 70 (HSP70), and high mobility group box 1 (HMGB1), IL-1 and several chemokines, then recruited T cells and acitived DC. The clinical value of ALA-PDT-induced antitumor responses in long-term control of SCCs deserves further study.

Protoporphyrin IX (PpIX) fluorescence imaging was conducted during routine dermatological methyl aminolevulinate-PDT. Linear and logistic regressions modelled any relationships between variables that may have affected PpIX accumulation/photobleaching and thus clinical outcome. Only reductions in photobleaching consistently adversely affected efficacy. Clinical clearance was reduced in lesions located on the limbs, hands and feet with lower PpIX accumulation and subsequent photobleaching adversely affecting outcomes. PpIX photobleaching was lower when air cooling pain relief was utilised, resulting in ~3-fold reduction in achieving clearance. PpIX photobleaching during the first treatment was concluded to be an excellent predictor of clinical outcome.

Photodynamic therapy (PDT) for squamous cell carcinoma (SCC) in situ continues to provide promising results in terms of its safety and efficacy. The most common indications are large or multiple lesions, poorly healing sites, and lesions in non-surgical patients. Current recommendations are the use of aminolevulinic (ALA) or methyl aminolevulinate (MAL), 3-4 hour incubation with a red light, 75 -100 J/cm, 2 to 3 sessions, 1-2 weeks apart. Compared to conventional therapy, MAL-PDT had a response rate of 93% vs cryotherapy at 86% and 5-Fluorouracil cream at 83%. Recurrence rates were higher than those seen with surgery. Overall, cosmetic scores were higher with PDT. These results indicate that PDT can be an effective treatment for SCC in situ. Prospective and randomized trials are needed to standardize PDT parameters and to obtain future approved indications.

The EU-approved protocols for photodynamic therapy of actinic keratosis require photoactivation with either red-light or daylight. Due to high irradiances, standard red-light photoactivation is painful. Several clinical studies have reported similar efficacy and better tolerability for red-light photoactivation at increasingly lower irradiances compared to standard red-light photoactivation. We have compared the irradiances measured during photoactivation with the Aktilite CL 128 (Galderma SA, Switzerland) with the associated complete responses at 3 months. The comparison purpose was to determine whether there is a minimum irradiance threshold for an effective red-light photoactivation. From this comparison, no such minimum irradiance threshold could be found.

Port-wine stains (PWS) are congenital vascular malformations, consisting of ectatic capillaries and post-capillary venules in the papillary and mid-reticular layers of the dermis. Although red PWS shows good therapeutic effect with laser, purple and hypertrophic PWS show tolerance which is associated with depth and diameter of vessels. We used dermascopy, VISIA, doppler ultrasound and two-dimensional (2D) ultrasound to evaluate treatment efficacy of PWS. Hypertrophic PWS showed reduction of blood flow by doppler ultrasound and attenuation of dermis and subcutaneous tissue by 2D ultrasound. Upon dermascopic examination, we found different types of PWS showed reduction of vessels and faded in color after treatment. In addition, VISIA was used to calculate the mutative area of erythema by treatment. Using non-invasive evaluation technique could be effective and scientific to evaluate the curative effect.

Long-term improvement of photoaging treated by topical ALA-PDT was studied via a split-face clinical study. Which means collagen in dermis was increased after ALA-PDT. Mechanism study aimed to investigate the molecular communication in ALA-PDT occurring between epidermal keratinocytes and dermal fibroblasts. Skin biopsies from mice were used to analyze the histological changes in dermal collagen and PpIX distribution. PpIX fluorescence was densely distributed in photoaged mouse epidermis while collagen in dermis underwent remodeling. Fibroblasts co-cultured with low-dose ALA-PDT showed ALA-PDT can stimulate keratinocytes to release TGF-β1, activating the TGF-β pathway in fibroblasts to remodel collagen and improve skin photaging.