There have been literature reports indicating that protocols involving two photosensitizing agents, in animal tumor models, can yield a synergistic result, i.e., more photokilling than can be obtained with either sensitizer alone at the same light dose. We have independently obtained similar results in a cell-culture system that permits a more detailed study of mechanisms involved. Using any of three agents that localize in lysosomes, we were able to show that low-dose lysosomal photodamage could substantially promote photokilling by benzoporphyrin derivative, an agent that primarily targets mitochondria. This effect was abolished by knockdowns of either of two genes normally associated with autophagy: ATG5 and ATG7. A mechanism that can account for these results is proposed.

The goal of this study was to develop and improve an infrared (IR) navigation system to deliver light dose uniformly during intracavitory PDT by tracking the movement of the light source and providing real-time feedback on the light fluence rate on the entire cavity surface area. In the current intrapleural PDT protocol, several detectors placed in selected locations in the pleural cavity monitor the light doses. To improve the delivery of light dose uniformity, an IR camera system is used to track the motion of the light source as well as the surface contour of the pleural cavity. Monte- Carlo simulation is used to improve the calculation algorithm for the effect of light that undergoes multiple scattering along the surface in addition to an improvement of the direct light calculation using an improved model that accounts for the anisotropy of the light from the light source.

Macroscopic modeling of the apparent reacted singlet oxygen concentration ([¹O₂]_rx) for use with photodynamic therapy (PDT) has been developed and studied for benzoporphryin derivative monoacid ring A (BPD), a common photosensitizer. The four photophysical parameters (ξ, σ, β, δ) and threshold singlet oxygen dose ([¹O₂]_{rx, sh}) have been investigated and determined using the RIF model of murine fibrosarcomas and interstitial treatment delivery. These parameters are examined and verified further by monitoring tumor growth post-PDT. BPD was administered at 1 mg/kg, and mice were treated 3 hours later with fluence rates ranging between 75 – 150 mW/cm² and total fluences of 100 – 350 J/cm². Treatment was delivered superficially using a collimated beam. Changes in tumor volume were tracked following treatment. The tumor growth rate was fitted for each treatment condition group and compared using dose metrics including total light dose, PDT dose, and reacted singlet oxygen. Initial data showing the correlation between outcomes and various dose metrics indicate that reacted singlet oxygen serves as a good dosimetric quantity for predicting PDT outcome.

The 2014 fiscal year (FY) continued to be a challenging one for all federal agencies despite the many Congressional strategies proposed to address the U.S. budget deficit. The Bipartisan Budget Act of 2013 passed by the House and Senate in December 2013 approved a two-year spending bill which cancelled the FY2014 and FY2015 required sequestration cuts (i.e., 4-5% National Institute of Health (NIH)/National Cancer Institute (NCI) budget reduction initiated on March 1, 2013), but extended the sequestration period through FY2023. This bill passage helped minimize any further budget reductions and resulted in a final FY2014 NIH budget of $29.9 billion and a NCI budget of $4.9 billion. Both NIH and NCI worked hard to maintain awarding the same number of NIH/NCI investigator-initiated R01 and exploratory R21 grants funded in FY2014 and similar to the level seen in FY2013 and previous years (see Tables 1 and 2). Since Congress only recently passed the 2015 spending bill in December 16, 2014, the final NIH and NCI budget appropriations for FY2015 remains unknown at this time and most likely will be similar to the FY2014 budget level. The NCI overall success and funding rates for unsolicited investigator-initiated R01 applications remained at 15%, while the success rate for exploratory R21 applications was 12% in FY2014 with similar rates seen in FY2013 (see Tables 1 and 2). The success rate for biomedical research applications in the Photodynamic Therapy and laser research field will be provided for the past few years. NIH provides numerous resources to help inform the extramural biomedical research community of new and current grant applicants about new grant policy changes and the grant submission and review processes.

Nonmelanoma skin cancer (NMSC), comprising basal cell carcinoma (BCC) and squamous cell carcinoma (SCC), is the most common form of human cancer worldwide. Effective therapies include surgical excision, cryotherapy, and ionizing radiation, but all of these cause scarring. ALA-based PDT is a non-scarring modality used routinely for NMSC in Europe but not in the USA, primarily due to lingering uncertainties about efficacy. We have identified three agents (methotrexate, 5-fluorouracil, and vitamin D) that can be used as neoadjuvants, i.e., can be given as a pretreatment prior to ALA-PDT, to improve the efficacy of tumor killing in mouse models of NMSC. Vitamin D (VD3) is the most recent neoadjuvant on this list. In this presentation we make the case that VD3 may be superior to the other agents to improve results of ALA-PDT skin cancer treatment. The active form of VD3 (calcitriol) is available topically as a pharmaceutical grade cream or ointment (FDA-approved for psoriasis), and works well for boosting ALA-PDT tumor treatment in mouse models. For deep tumors not reachable by a topical route, calcitriol can be given systemically and is very effective, but carries a risk of causing hypercalcemia as a side effect. To circumvent this risk, we have conducted experiments with the natural dietary form of VD3 (cholecalciferol), and showed that this improves ALA-PDT efficacy almost to the same extent as calcitriol. Because cholecalciferol does not increase serum calcium levels, this represents a potentially extremely safe approach. Data in mouse models of BCC and SCC will be presented.

Interstitial photodynamic therapy (iPDT) describes the use of implanted optical fibers for delivery of treatment light to activate photosensitizer in regions that can be located deep within the body. Since sensitive healthy structures are often located nearby, this requires careful treatment planning that is dependent on tissue optical properties. Determination of these values usually involves the insertion of additional fibers into the volume, or the use of flat-cleaved optical fibers as both treatment sources and detectors. The insertion of additional fibers is undesirable, and cylindrical diffusers have been shown to offer superior treatment characteristics compared to flat-cleaved fibers. Using cylindrical diffusers as detectors for spectroscopic measurement is therefore attractive. We describe the determination of the detection profile for a particular cylindrical diffuser design and derive the scatterer concentration gradient within the diffuser core. This detection profile is compared to previously characterized diffusers, and is shown to be dependent on the diffuser design. For diffusers with a constant scatterer concentration and distal mirror, the detection profile is localized to the proximal end of the diffusing region. For diffusers with variable scattering concentration along their length and no distal mirror, the detection profile is shown to be more uniform along the diffusing region. We also present preliminary results showing the recovery of optical properties using arrays of cylindrical diffusing fibers as sources and detectors, with a mean error of 4.4% in the determination of μ_eff. The accuracy of these results is comparable to those obtained with other methods of optical property recovery.

Photodynamic therapy (PDT) is a treatment modality in which cytotoxic reactive oxygen species are generated from oxygen and other biological molecules when a photosensitizer is activated by light. PDT has been approved for the treatment of cancers and age-related macular degeneration (AMD) due to its effectiveness in cell killing and manageable normal tissue complications. In this study, we characterized the effects of verteporfin-PDT on SVEC mouse endothelial cells and determined its underlying cell death mechanisms. We found that verteporfin was primarily localized in mitochondria and endoplasmic reticulum (ER) in SVEC cells. Light treatment of photosensitized SVEC cells induced a rapid onset of cell apoptosis. In addition to significant structural damages to mitochondria and ER, verteporfin-PDT caused substantial degradation of ER signaling molecules, suggesting ER stress. These results demonstrate that verteporfin-PDT triggered SVEC cell apoptosis by both mitochondrial and ER stress pathways. Results from this study may lead to novel therapeutic approaches to enhance PDT outcome.

A novel series of poly (aryl benzyl ether) dendrimer silicon phthalocyanines loaded block copolymers ethoxypoly(ethylene glycol)-poly (lactic-co-glycolic acid) (MPEG-PLGA)were formed. The time-dependent intracellular uptake of nanoparticles in HUVECs cells increased as they were incorporated into nanoparticles. With its highly effective selective accumulation on choroidal neovascularization(CNV). This treatment resulted in a efficacious choroidal neovascularization (CNV) occlusion with minimal unfavorable phototoxicity.

Type II photodynamic therapy (PDT) is based on the use of photochemical reactions mediated through an interaction between a tumor-selective photosensitizer, photoexcitation with a specific wavelength of light, and production of reactive singlet oxygen. However, the medical application of this technique has been limited due to inaccurate PDT dosimetric methods. The goal of this study is to examine the relationship between outcome (in terms of tumor growth rate) and calculated reacted singlet oxygen concentration [¹O₂]_rx after HPPH-mediated PDT to compare with other PDT dose metrics, such as PDT dose or total light fluence. Mice with radiation-induced fibrosarcoma (RIF) tumors were treated with different light fluence and fluence rate conditions. Explicit measurements of photosensitizer drug concentration and tissue optical properties via fluorescence and absorption measurement with a contact probe before and after PDT were taken to then quantify total light fluence, PDT dose, and [¹O₂]_rx based on a macroscopic model of singlet oxygen. In addition, photobleaching of photosenitizer were measured during PDT as a second check of the model. Changes in tumor volume were tracked following treatment and compared to the three calculated dose metrics. The correlations between total light fluence, PDT dose, reacted [¹O₂]_rx and tumor growth demonstrate that [¹O₂]_rx serves as a better dosimetric quantity for predicting treatment outcome and a clinically relevant tumor growth endpoint.

Photodynamic therapy (PDT) delivers a localized cytotoxic dose that is a function of tissue oxygen availability, photosensitive drug concentration, and light fluence. Providing safe and effective PDT requires an understanding of all three elements and the physiological response to the radicals generated. Interstitial PDT (IPDT) for solid tumours poses particular challenges due to complex organ geometries and the associated limitations for diffusion theory based fluence rate prediction, in addition to restricted access for light delivery and dose monitoring.

As a first step towards enabling a complete prospective IPDT treatment-planning platform, we demonstrate use of our previously developed FullMonte tetrahedral Monte Carlo simulation engine for modeling of the interstitial fluence field due to intravesicular insertion of brief light sources. The goal is to enable a complete treatment planning and monitoring work flow analogous to that used in ionizing radiation therapy, including plan evaluation through dose-volume histograms and algorithmic treatment plan optimization.

FullMonte is to our knowledge the fastest open-source tetrahedral MC light propagation software. Using custom hardware acceleration, we achieve 4x faster computing with 67x better power efficiency for limited-size meshes compared to the software. Ongoing work will improve the performance advantage to 16x with unlimited mesh size, enabling algorithmic plan optimization in reasonable time.

Using FullMonte, we demonstrate significant new plan-evaluation capabilities including fluence field visualization, generation of organ dose-volume histograms, and rendering of isofluence surfaces for a representative bladder cancer mesh from a real patient. We also discuss the advantages of MC simulations for dose-volume histogram generation and the need for online personalized fluence-rate monitoring.

Non-melanoma skin cancers (NMSCs) such as basal cell carcinoma (BCC) and squamous cell carcinoma (SCC) are the most common form of human cancer worldwide, and their incidence is increasing. Photodynamic therapy (PDT), mediated by topically applied aminolevulinic acid (ALA) and subsequent exposure to light (either a laser or a noncoherent source), is being increasingly used for the treatment of dermatological disorders, including BCC and SCC. However, therapeutic responses of NMSCs to ALA-PDT are currently not superior to standard therapies, although the latter have undesirable side effects including scarring. In this study, we report that preconditioning of skin tumors with calcitriol (active form of Vitamin D; Vit D) prior to ALA-PDT, significantly improves the treatment outcome. In BCC and UVB-induced SCC mouse models, we identified an increase in tumor-specific accumulation of ALA induced photosensitizer (protoporphyrin IX, PpIX) due to Vit D preconditioning, of up to 6- fold in vivo. In addition, increased expression of differentiation (145 fold, p < 0.02) and proliferation (42 fold, p <0.005) markers were identified in BCC tumors, all leading to increased tumor destruction (18.3 fold, p < 0.03) with the combination approach, as compared to ALA-PDT alone. Histomorphological changes identified using hematoxylin and eosin staining, and results of TUNEL staining, together documented a beneficial effect of Vit D pretreatment upon tumor cell death. We conclude that this new combination approach with Vit D and ALA-PDT has great potential to achieve complete remission of NMSC tumors, with excellent cosmetic results and an overall beneficial impact upon patient care.

Nanoparticles made from aluminum phthalocyanine (AlPc) raw material are non-fluorescent because of fluorescence quenching due to the molecular crystalline structure forming a stack of flat molecular layers. However, when AlPc molecules become detached from the particle, fluorescence occurs. First observations demonstrated the benefit of using aluminum phthalocyanine nanoparticles (nAlPc) for the assessment of the rejection risk of skin autografts in mice by measuring fluorescence intensities of detached AlPc molecules. Skin autografts showing a high fluorescence intensity of AlPc were finally rejected induced by an inflammatory process. In contrast, autografts with normal skin autofluorescence were accepted. This finding stimulated our work to reveal the mechanism of the AlPc fluorescence development from the nanoparticles. This could be used to specifically detect inflammatory processes or tumors and will have the potential of using nAlPc as a new treatment modality for PDT.

The purpose of this study is to develop an alternate empirical approach to estimate near-infra-red (NIR) photon propagation and quantify optically induced drug release in brain metastasis, without relying on computationally expensive Monte Carlo techniques (gold standard). Targeted drug delivery with optically induced drug release is a noninvasive means to treat cancers and metastasis. This study is part of a larger project to treat brain metastasis by delivering lapatinib-drug-nanocomplexes and activating NIR-induced drug release. The empirical model was developed using a weighted approach to estimate photon scattering in tissues and calibrated using a GPU based 3D Monte Carlo. The empirical model was developed and tested against Monte Carlo in optical brain phantoms for pencil beams (width 1mm) and broad beams (width 10mm). The empirical algorithm was tested against the Monte Carlo for different albedos along with diffusion equation and in simulated brain phantoms resembling white-matter (μ_s’=8.25mm^-1, μ_a=0.005mm^-1) and gray-matter (μ_s’=2.45mm^-1, μ_a=0.035mm^-1) at wavelength 800nm. The goodness of fit between the two models was determined using coefficient of determination (R-squared analysis). Preliminary results show the Empirical algorithm matches Monte Carlo simulated fluence over a wide range of albedo (0.7 to 0.99), while the diffusion equation fails for lower albedo. The photon fluence generated by empirical code matched the Monte Carlo in homogeneous phantoms (R²=0.99). While GPU based Monte Carlo achieved 300X acceleration compared to earlier CPU based models, the empirical code is 700X faster than the Monte Carlo for a typical super-Gaussian laser beam.

The intermolecular electron transfer between the novel dendritic zinc (II) phthalocyanines (G₁-DPcB and G₂-DPcB) and anthraquinone (AQ) was studied by steady-state fluorescence and UV/Vis absorption spectroscopic methods. The effect of dendron generation on intermolecular electron transfer was investigated. The results showed that the fluorescence emission of these dendritic phthalocyanines could be greatly quenched by AQ upon excitation at 610 nm. The Stern- Volmer constant (K_SV) of electron transfer was decreased with increasing the dendron generations. Our study suggested that these novel dendritic phthalocyanines were effective new electron donors and transmission complexes and could be used as a potential artifical photosysthesis system.

A macroscopic singlet oxygen model for photodynamic therapy (PDT) has been used extensively to calculate the reacted singlet oxygen concentration for various photosensitizers. The four photophysical parameters (ξ, σ, β, δ) and threshold singlet oxygen dose ([¹O₂]_r,sh) can be found for various drugs and drug-light intervals using a fitting algorithm. The input parameters for this model include the fluence, photosensitizer concentration, optical properties, and necrosis radius. An additional input variable of photobleaching was implemented in this study to optimize the results. Photobleaching was measured by using the pre-PDT and post-PDT sensitizer concentrations. Using the RIF model of murine fibrosarcoma, mice were treated with a linear source with fluence rates from 12 - 150 mW/cm and total fluences from 24 – 135 J/cm. The two main drugs investigated were benzoporphyrin derivative monoacid ring A (BPD) and 2-[1-hexyloxyethyl]-2-devinyl pyropheophorbide-a (HPPH). Previously published photophysical parameters were fine-tuned and verified using photobleaching as the additional fitting parameter. Furthermore, photobleaching can be used as an indicator of the robustness of the model for the particular mouse experiment by comparing the experimental and model-calculated photobleaching ratio.

In this study, effect of a novel LED-based light source developed for 96-well-plates cell culture applications, was tried on AGS stomach cancer cell line, in combination with Poly(amido amine) (PAMAM) modified – porhyrin molecule. For each 4 generation of modified PpIX molecule 5 different concentrations tried. According to results PAMAM molecule doesnt have any photosensitizer property also didn’t show any toxic effect even if higher concentrations. Morphology and real time monitoring analysis results hold up each other and confirmed that, PpIX molecules with and without modificated high concentrations (>100μM) caused cell death via toxicicity this reason optimal concentration for PAMAM modified PpIX should be between 25 - 50 μm concentration .

Photodynamic therapy (PDT) has shown promising results in targeted treatment of cancerous cells by developing localized toxicity with the help of light induced generation of reactive molecular species. The efficiency of this therapy depends on the product of the intensity of light dose and the concentration of photosensitizer (PS) in the region of interest (ROI). On account of this, the dynamic and variable nature of PS delivery and retention depends on many physiological factors that are known to be heterogeneous within and amongst tumors (e.g., blood flow, blood volume, vascular permeability, and lymph drainage rate). This presents a major challenge with respect to how the optimal time and interval of light delivery is chosen, which ideally would be when the concentration of PS molecule is at its maximum in the ROI. In this paper, a predictive algorithm is developed that takes into consideration the variability and dynamic nature of PS distribution in the body on a region-by-region basis and provides an estimate of the optimum time when the PS concentration will be maximum in the ROI. The advantage of the algorithm lies in the fact that it predicts the time in advance as it takes only a sample of initial data points (~12 min) as input. The optimum time calculated using the algorithm estimated a maximum dose that was only 0.58 ± 1.92% under the true maximum dose compared to a mean dose error of 39.85 ± 6.45% if a 1 h optimal light deliver time was assumed for patients with different efflux rate constants of the PS, assuming they have the same plasma function. Therefore, if the uptake values of PS for the blood and the ROI is known for only first 12 minutes, the entire curve along with the optimum time of light radiation can be predicted with the help of this algorithm.

We investigated the optical properties of novel terbium (Tb³⁺)-doped nanophosphors with various host compounds irradiated by clinical electron, photon, and proton beams for their potential as optical probes. The emission spectra of nanophosphors embedded in tissue-mimicking phantoms were collected by an optical fiber connected to a CCD-coupled spectrograph while the samples were irradiated with electron and photon beams generated by a medical linear accelerator and proton beams generated by a clinical cyclotron. We characterized the luminescence of such nanophosphors as a function of the beam energy and observed a dose dependency of the luminescence signal. We demonstrated x-ray luminescence, cathodoluminescence, and ionoluminescence of the nanophosphors in clinical ionizing radiation fields, which indicates their potential as downconverters of high-energy radiation into visible light.

In this study, copper chlorophyllin was used as a photosensitizer for photodynamic therapy (PDT) in Ehrlich tumour mouse model. Six groups of animals comprising 5 animals per group were subcutaneously transplanted with 1x10⁶ Ehrlich tumour cells. A single dose of 200 μg/gm body weight chlorophylin derivative was administered by intravenous (IV) or intratumoral (IT) routes. Mice were exposed to monochromatic red laser of 630 nm for 1 h, and tumour regression was followed up for three consecutive months post treatment. Several Biochemical, histological and molecular tests were performed in order to evaluate the efficacy and safety of the applied treatment. An interest has been also directed towards investigating the molecular mechanisms underlying chlorophyllin derivative mediated PDT. PDT-treated animals via either the IV or IT routes showed significant decrease in tumour size 72 h post-treatment. Tumours at the IV-PDT group disappeared totally within a week with no recurrence over three months follow up. In the IT-PDT, the decrease in tumour size at the first week was interrupted by a slight increase; however never reached the original size. Histological examination of tumour tissues of the IV-PDT group at 24 h post treatment demonstrated restoring the normal muscle tissue architecture, and the biochemical assays indicated normal liver functions. The immunohistochemical analysis of caspase-3, and the quantitative PCR results of caspases-8 and 9 proved the presence of extrinsic apoptotic pathway after cholorphyllin derivative-mediated PDT. In conclusion IV-PDT strategy proved better cure rate than the IT-PDT, with no recurrence over three months of follow up.

Traditional treatment options for prostate cancer are insufficient to cure advanced drug-resistant prostate cancer. Thus, as an alternative form of cancer therapy, photodynamic therapy (PDT) has become the main subject of intense investigation as a possible treatment modality. In this study, ultraviolet-inactivated viral vector, called hemagglutinating virus of Japan envelope (HVJ-E) was utilized to establish an effective delivery system for photosensitizer. Lipidated protoporphyrin IX (PpIX lipid) was inserted in HVJ-E by centrifugation to create a new drug delivering system that allows selective accumulation of photosensitizers in cancer cells. To study in vitro drug release mechanism of porphyrus envelope, the ultra-high voltage electron microscope tomography was applied. Next, to evaluate the photodynamic efficiency of porphyrus envelope for hormone antagonistic prostate cancer cells (PC-3), uptake of porphyrus envelope derived PpIX lipid and PpIX induced from exogenously administered precursor of 5-aminolevulinic acid hydrochloride (5-ALA) were compared by measuring fluorescence intensity of PpIX. Finally, to evaluate the efficacy of porphyrus envelope-PDT, laser light at a wavelength of 405 nm was irradiated to PC-3 cells. As a result, incorporation of porphyrus envelope-derived PpIX lipid occurred via membrane fusion, giving the highest fluorescence intensity when compared to 5-ALA-induced PpIX. Also, results from PDT experiment revealed the 28.6 × 10³-fold and 206-fold increase in therapeutic efficacy when compared to those of PDT using 5-ALA induced PpIX and PpIX lipid, respectively. Our findings suggest how porphyrus envelope can induce efficient accumulation of PpIX lipid, which can enhance the therapeutic efficacy of PDT against hormone antagonistic prostate cancer.