This PDF file contains the front matter associated with SPIE Proceedings Volume 8582, including the Title Page, Copyright information, Table of Contents, and the Conference Committee listing.

We have previously shown that photodynamic therapy mediated by a vascular regimen of benzoporphyrin derivative and 690nm light is capable of inducing a robust immune response in the mouse CT26.CL25 tumor model that contains a tumor-rejection antigen, beta-galactosidase (β-gal). For the first time we show that PDT can stimulate the production of serum IgG antibodies against the β-gal antigen. It is known that a common cause of death from cancer, particularly lung cancer, is brain metastases; especially the inoperable ones that do not respond to traditional cytotoxic therapies either. We asked whether PDT of a primary tumor could stimulate immune response that could attack the distant brain metastases. We have developed a mouse model of generating brain metastases by injecting CT26.CL25 tumor cells into the brain as well as injecting the same cancer cells under the skin at the same time. When the subcutaneous tumor was treated with PDT, we observed a survival advantage compared to mice that had untreated brain metastases alone.

Objectives: To examine therapeutic effects of 5-aminolevulinate (ALA)-mediated photodynamic therapy (PDT) on UVB-induced cutaneous squamous cell carcinomas (SCCs) in a mouse model. Materials and methods: Cutaneous SCCs were established by UVB (280-320 nm) irradiation of hairless mice. In situ fluorescence measurement was used to monitor PpIX formation after the topical application of various concentrations of ALA cream to determine the optimal ALA dose. Therapeutic responses of SCCs to multiple sessions of ALA PDT were examined histologically and quantitatively. TUNEL staining was used to examine apoptosis caused by PDT. Results: After repeated exposure for 18 to 22 weeks (4-5 days/week), multiple nodular and verrucous hyperplasia lesions of various sizes developed at the exposed area. After four sessions of ALA PDT (8% ALA, 3 h incubation, 30 J/cm² at 20 mW/cm²) a total of 84% of complete response was achieved for small SCCs (1-4 mm, thickness <2.5 mm). TUNEL staining showed that PDT-induced apoptotic cells were distributed evenly from the basal to stratum corneum layers. Conclusions: Topical ALA PDT can trigger apoptosis in SCCs, inhibit SCC growth, and reduce the size and number of tumors in the hairless mouse model. The true clinical value of ALA PDT for the treatment of cutaneous SCC deserves further investigation.

Laser immunotherapy (LIT) uses laser irradiation and immunological stimulation to treat metastatic cancers. The current mode of operation of LIT is through dye-enhanced non-invasive irradiation. Although this treatment has given promising results, there are still a number of challenges with this method, such as limited light penetration for deep tumors and strong light absorption by highly pigmented skins. Interstitial laser immunotherapy (ILIT), using a cylindrical diffuser, is designed to overcome these limitations. In this study, rat tumors were treated by ILIT with an 805 nm laser and varying doses of glycated chitosan, an immunological stimulant. The goal was to observe the effects of differing doses of the stimulant on the survival of the tumor-bearing rats. The results suggested that the optimal dose of glycated chitosan is in the range of 0.1 to 0.3 ml per rat tumor.

Combining near infrared (NIR) laser irradiation into a tumor treatment therapy has shown promising results. For a comprehensive tumor therapy, it is important to understand the effects of NIR irradiation not only on the tumor, but on the tumor stroma as well. The composition of the microenvironment present near the tumor cells is critical to the phenotype of the tumor. Fibroblasts affect tissue homeostasis and change the microenvironment surrounding the tumor. Myofibroblast are derived from fibroblast cells, and in some cases indicate the transformation of healthy tissue into malignant tissue. Wound healing environments are rich in fibroblast cells and are similar to tumor stromas. To simulate a tumor stroma a wound healing environment was constructed. Two different human fibroblast cells were cultured in collagen lattices. Specifically, collagen lattices were created, with type 1 collagen, incubated for 5 days and irradiated with a 980nm laser on the 4th day. The subsequent collagen lattices were either released and measured, or fixed for immunostaining on the 5th day; the contraction rates also were analyzed. Furthermore, collagen lattices were stained to identify fibroblast proliferation and differentiation, into myofibroblasts. The results suggested NIR laser irradiation had some biological effects on the fibroblast cells, but the full extent of the effects is still unclear.

In this study, we proposed photoacoustic imaging to monitor the temperature increase during photothermal therapy, which was carried out by utilizing a continuous wave laser and photoabsorber-enhanced black ink with a absorption peak in the near-infrared optical range. A focusing photoacoustic imaging is interfaced with a nanosecond pulsed laser to image tissue-mimicking phantoms before and after laser irradiation. The results demonstrated that changes in the photoacoustic signal could reflect temperature changes in tissue. More importantly, photoacoustic signal could be used to determine the temperature at the boundary of photoabsorber-enhance tissue during photothermal irradiation. Thus, the photoacoustic imaging could potentially become an effective tool to guide photoabsorber-enhanced photothermal therapy.

In this study, we have prepared tumor-targeted hydrophilic UCNPs by coating folate-modified amphiphilic SOC polymer on the surface of OA-UNCPs through hydrophobic interaction. The produced FASOC-UCNPs have excellent optical properties and good dispersibility in aqueous solution. TEM image and FTIR spectra confirm the successful surface modification of OA-UCNPs and folate-conjugation on the UCNPs. Folate-conjugated UCNPs possess active-targeting ability mediated by FR, compared with passive-targeted SOC-UCNP taken up by tumors via the EPR effect. Singlet oxygen production in cancer cells induced by FASOC-UCNP-ZnPc demonstrates the great potential of the nanocomplex as a novel PDT agent based on UCNPs.

We previously reported on the synergistic effects of hyperthermia and chemotherapy using doxorubicin (DOX) and Indocyanine Green (ICG). In a previous study we also explored the potential use of simultaneous entrapment of optical/imaging and chemotherapeutic agents into PLGA nanoparticles. The aim of the present study is to further decorate their surface with tumor specific monoclonal antibodies in order to achieve simultaneous therapy and diagnosis in a targeted manner. Thus, ICG was selected as an imaging agent due to its wide clinical applications and since it can also serve as hyperthermia agent. DOX was selected as the chemotherapeutic agent since it is used clinically for a large spectrum of tumors.

Carbon nanotubes have a great potential in the biomedical applications. To use carbon nanotubes in the treatment of cancer, we synthesized an immunologically modified single-walled carbon nanotube (SWNT) using a novel immunomodifier, glycated chitosan (GC), as an effective surfactant for SWNT. This new composition SWNT-GC was stable due to the strong non-covalent binding between SWNT and GC. The structure of SWNT-GC is presented in this report. The photothermal effect of SWNT-GC was investigated under irradiation of a near-infrared laser. SWNT-GC retained the optical properties of SWNT and the immunological properties of GC. Specifically, the SWNT-GC could selectively absorb a 980-nm light and induce desirable thermal effects in tissue culture and in animals. It could also induce tumor cell destruction, controlled by the laser settings and the doses of SWNT and GC. Laser+SWNT-GC treatment could also induce strong expression of heat shock proteins on the surface of tumor cells. This immunologically modified carbon nanotube could be used for selective photothermal interactions in noninvasive tumor treatment.

Hepatocellular carcinoma (HCC) is one of the most common malignancies in the world, with approximately 1,000,000 cases reported every year. The fate of circulating tumor cells (CTCs) is an important determinant of metastasis and recurrence, which lead to most deaths in HCC. Therefore, quantification of CTCs proves to be an emerging tool for diagnosing, stratifying and monitoring patients with metastatic diseases. In vivo flow cytometry (IVFC) has the capability to monitor the dynamics of fluorescently labeled CTCs continuously and non-invasively. Here, we combine IVFC technique and a GFP-transfected HCC orthotopic metastatic tumor model to monitor CTC dynamics. Our IVFC has ~1.8-fold higher sensitivity than whole blood analysis by conventional flow cytometry. We find out a significant difference of CTC dynamics between orthotopic and subcutaneous (s.c.) tumor models. We also investigate whether liver resection promotes or restricts hematogenous metastasis in advanced HCC. Our result shows that the number of CTCs and early metastases decreases after the resection. CTC dynamics is correlated with tumor growth in our orthotopic tumor model. The number and size of distant metastases correspond to CTC dynamics. The novel IVFC technique combined with orthotopic tumor models might provide insights to tumor hematogenous metastasis and guidance to cancer therapy.

The purpose of this study was to determine the Contrast to Noise Ratio (CNR) of the x-ray images taken with the phase contrast imaging mode and compare them with the CNR of the images taken under the conventional mode. For each mode, three images were taken under three exposure conditions of 100 kVp (2.8mAs), 120 kVp (1.9mAs) and 140kVp (1.42mAs). A 1.61cm thick contrast detail phantom was used as an imaging object. For phase contrast, the source to image detector distance (SID) was 182.88 cm and the source to object (SOD) distance was 73.15 cm. The SOD was the same as SID in the conventional imaging mode. A computed radiography (CR) plate was used as a detector and the output CR images were converted to linear form in relation with the incident x-ray exposure. To calculate CNR, an image processing software was used to determine the mean pixel value and the standard deviation of the pixels in the region of interest (ROI) and in the nearby background around ROI. At any given exposure condition investigated in this study, the CNR values for the phase contrast images were better as compared to the corresponding conventional mode images. The superior image quality in terms of CNR is contributed by the phase-shifts resulted contrast, as well as the reduced scatters due to the air gap between the object and the detector.

The purpose of this study is to objectively investigate image sharpness of the metaphase chromosomes at different scanning speeds. In cytogenetic imaging, high scanning speed is sometimes applied to optimize the efficiency. However, at high scanning speed, the obtained image may be deteriorated, due to the scanning blur, under exposure and random vibration of the scanning stage. In this study, the image sharpness of metaphase chromosomes is objectively evaluated. A standard resolution target and several metaphase chromosomes are first imaged under different speeds, by a prototype scanning microscope. Then the sharpness of the acquired images is objectively assessed using a sharpness function. For this prototype system, the results demonstrate that the image sharpness of the metaphase chromosomes is optimized when the scanning speed is 0.8mm/s, under the specified experimental conditions. The results of this study may be useful for optimizing the efficiency and quality of clinical cytogenetic imaging procedures.

This research is aimed at studying the advantages of an x-ray phase-contrast tomosynthesis prototype by using phantoms. A prototype system is assembled with a micro-focus x-ray source, a rotating stage and a computed radiography detector mounted on an optical rail. A custom designed bubble wrap phantom is used in experiments. Angular projection images are acquired from -20° to +20° with 2° interval. The in-plane slices are reconstructed. The feature area on the phantom is observed. The prototype system provides an intrinsic way to investigate the potential and imaging quality of a phase-contrast tomosynthesis imaging method. As the result, phase-contrast tomosynthesis imaging method is demonstrated for its advantages in avoiding structure noise and overlapping issues by comparing the results acquired by computed radiography and phase-contrast radiography.

While successes of different cancer therapies have been achieved in various degrees a systemic immune response is needed to effectively treat late-stage, metastatic cancers, and to establish long-term tumor resistance in the patients. A novel method for combating metastatic cancers has been developed using immunologically modified carbon nanotubes in conjunction with phototherapy. Glycated chitosan (GC) is a potent immunological adjuvant capable of increasing host immune responses, including antigen presentation by activation of dendritic cells (DCs) and causing T cell proliferation. GC is also an effective surfactant for nanomaterials. By combining single-walled carbon nanotubes (SWNTs) and GC, immunologically modified carbon nanotubes (SWNT-GC) were constructed. The SWNT-GC suspension retains the enhanced light absorption properties in the near infrared (NIR) region and the ability to enter cells, which are characteristic of SWNTs. The SWNT-GC also retains the immunological properties of GC. Cellular SWNT-GC treatments increased macrophage activity, DC activation and T cell proliferation. When cellular SWNT-GC was irradiated with a laser of an appropriate wavelength, these immune activities could be enhanced. The combination of laser irradiation and SWNT-GC induced cellular toxicity in targeted tumor cells, leading to a systemic antitumor response. Immunologically modified carbon nanotubes in conjunction with phototherapy is a novel and promising method to produce a systemic immune response for the treatment of metastatic cancers.

The actin depolymerizing factor (ADF)/cofilin protein family has been reported to be associated with ischemia induced renal disorders. Here we examine if cofilin-1 is associated with acute kidney injury (AKI). We exploited a 96-well based fiber-optic biosensor that uses conjugated gold nanoparticles and a sandwich immunoassay to detect the urine cofilin-1 level of AKI patients. The mean urine cofilin-1 level of the AKI patients was two-fold higher than that of healthy adults. The receiver operating characteristic (ROC) curve showed that cofilin-1 is a potential biomarker for discriminating AKI patients from healthy adults for intensive care patients.

Osteoarthritis (OA) is characterized by a slowly progressing degradation of the matrix and destruction of articular cartilage. Apoptosis of chondrocyte is accounted for the mechanism of OA. Nitric oxide (NO), as a stimulus, has been shown to induce chondrocyte apoptosis by activating the matrix metalloproteinases (MMPs), increasing the expression of cyclooxygenase 2 (COX-2) and the level of prostaglandin E2 (PGE2), inhibiting the proteoglycan synthesis and type II collagen expression. In this study, sodium nitroprusside (SNP) was administered to be the NO donor to explore the mechanism of NO-induced apoptosis of rabbit chondrocytes obtained from six weeks old New Zealand rabbits. CCK-8 assay revealed the inhibitory effect of SNP on cell viability. We used flow cytometry (FCM) to assess the form of cell death by Annexin-V/propidium iodide (PI) double staining, and evaluate the change of mitochondrial membrane potential (ΔΨm). We found that the SNP induced chondrocyte apoptosis in a dose- and time-dependent manner and an observable reduction of ΔΨm. In conclusion, our findings indicate that SNP induces apoptosis of rabbit chondrocytes via a mitochondria-mediated pathway.

Thermal therapy is based on the observation that tumor cells are sensitive to increased temperature, which is important for tumor control. In this study, the high intensity focused ultrasound (HIFU) system was used to simulate thermal therapy on breast cancer control in the small animal model. Additionally, the immunoadjuvant agent, so called glycated chitosan (GC), was used to enhance the immunological effects on tumor control. The bioluminescent imaging showed that tumor metastasis was apparently suppressed by a combined treatment using HIFU and GC, but not in HIFU or GC alone. Using immunohistochemical (IHC) staining, lung metastasis of 4T1-3R tumor cells further agree the observations obtained from non-invasive in vivo imaging. We also found that plasma collected from mice treated with combined HIFU and GC could significantly suppress the viability of cultured 4T1 cells compared to untreated or single treated group. In summary, these results suggest that the HIFU therapy combined with GC can enhance the tumor immunogenicity and tumor control.

Hepatocarcinoma, a malignant cancer, threaten human life badly. It is a current issue to seek the effective natural remedy from plant to treat cancer due to the resistence of the advanced hepatocarcinoma to chemotherapy. In this report, we assessed the antitumor activity of a prunellae spica aqueous extract (PSE) in vitro and in vivo. PSE was quantified by HPLC and UV. MTT assay showed that PSE did not effectively inhibit the growth of H22 cells. The in vivo anti-tumor activity was assessed by using the mice bearing H22 tumor. In vivo studies showed the higher antitumor efficacy of PSE without significant side effect assessed by the reduced tumor weight, and the extended survival time of the mice bearing H22 solid and ascites tumor. Collectively, PSE is a promising Chinese medicinal herb for treating hepatocarcinoma.

The epidermal growth factor receptor EGFR and HER2 are members of recepeter tyrosine kinase family. Overexpression of EGFR and HER2 has been observed in a variety of human tumors, making these receptors promising targets for tumor diagnosis. An affibody targeting HER2 and a nanobody targeting EGFR were reported before. In this Manuscript, we described an bispecific peptide combined with an affibody and a nanonbody through a linker―（G4S）3 . And the bispecific peptide was labeled with near-infrared (NIR) fluorochrome ICG-Der-02 for in vivo tumor EGFR and HER2 targeting. Afterwards, the EGFR and HER2 specificity of the fluorescent probe was tested in vitro for receptor binding assay and fluorescence microscopy and in vivo for subcutaneous MDA-MB-231 tumor targeting. The results indicated that the bispecific peptide had a high affinity to EGFR and HER2. Besides, in vitro and in vivo tumor targeting experiment indicated that the ICG-Der-02-( bispecific peptide) showed excellent tumor activity accumulation. Noninvasive NIR fluorescence imaging is able to detect tumor EGFR and HER2 expression based upon the highly potent bispecific peptide probe.

Asialoglycoprotein receptors distributing on the surface of hepatic parenchymal cells could specifically recognize galactose residues. Therefore, galactose residues can be applied in hepatocellar carcinoma targeted therapy through binding to the asialoglycoprotein receptors. A drug carrier containing of near infrared (NIR) fluorescence dye (MPA) and galactose was constructed by covalently conjugation process and named as GAL-MPA. The optical properties, cell viability, cell affinity and in vivo bio-distribution of this drug carrier were evaluated respectively. The results demonstrated that the drug carrier was provided with NIR fluorescence after conjugated with MPA. The low bio-toxicity and high hepatic cells affinity facilitated this drug carrier for in vivo biomedical application. Finally, the in vivo dynamic distribution of GAL-MPA confirmed this drug carrier was a promising candidate for tumor-targeting imaging and therapy.

Harmine is a beta-carboline alkaloid from the plant Peganum harmala. These alkaloids were stimulated by their promising antitumor activities in the recent years. In this study, we designed and synthesized two harmine derivatives #1and #2 modified at position-9 of harmine with ethyl and phenylpropyl, respectively. To improve the tumor targeting capability, #1’ and #2’ were synthesized by conjugating 2-amino-2-deoxy-D-glucose (2DG) to the derivatives #1 and #2, respectively. The MTT assays of all these compounds in vitro against L02, HepG2 showed all compounds had low toxicity to normal cells (L02) and significantly enhanced carcinoma cell inhibitory rate compared to harmine. Cytotoxicity against liver cancer cell lines of compound #1’ #2’ is higher than #1 #2, and even the compound #2’ is better than positive drug 5-FU. The compound #2’, a novel 2DG-based harmine derivatives, could become a promising drug for targeted cancer therapy and combination therapy with other antitumor drugs.

Macrophages appear to be directly involved in cancer progression and metastasis. However, the role of macrophages in influencing tumor metastasis has not been fully understood. Here, we have used an emerging technique, namely in vivo flow cytometry (IVFC) to study the depletion kinetics of circulating prostate cancer cells in mice and how depletion of macrophages by the liposome-encapsulated clodronate affects the depletion kinetics. Our results show different depletion kinetics of PC-3 prostate cancer cells between macrophage-deficient group and the control group. The number of circulating tumor cells (CTCs) in macrophage-deficient group decreases in a slower manner compared to the control mice group. The differences in depletion kinetics indicate that the absence of macrophages might facilitate the stay of prostate tumor cells in circulation. We speculate that macrophages might be able to arrest, phagocytose and digest PC-3 cancer cells. Therefore, the phagocytosis may mainly contribute to the differences in depletion kinetics. The developed methods here would be useful to study the relationship between macrophages and cancer metastasis in small animal tumor model.

High fluence low-power laser irradiation (HF-LPLI) provides a new stimulator to trigger cell apoptosis, and it is well known that apoptotic cells provide antigens to effectively trigger recognition by the immune system. In order to investigate the effect of HF-LPLI on the professional antigen-presenting cell (APC) function, in our primary study, we focused our attention on the effect of HF-LPLI-treated tumor cells on macrophages phagocytosis and NO production. Both confocal microscopy and flowcytometry analysis showed that HF-LPLI (120 J/cm²) induced significantly EMT6 death. Further experiments showed that HF-LPLI-treated EMT6 cells could be phagocyted by the murine macrophage cells RAW264.7, and could induce NO production in macrophages. Taken together, our results indicate that HF-LPLI-treated tumor cells effectively regulated the immune system. The HF-LPLI effect on the APC function needs to be further studied.