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Int. J. Mol. Sci. 2018, 19, 1107 3 of 17 Recently Fan et al. [29] reported about the development of MB bound nanoplatform, which is capable of delivering targeted diagnostic and photodynamic treatment of cancer. Once the nanoparticle binds with the target cell surface, it can detect human prostate cancer cell selectively using fluorescence imaging and PDT treatment using 785 nm, near infrared light indicates that the multimodal treatment increases the possibility of destroying prostate cancer cells in vitro [29]. 1.3. In Vitro Data There exist various in vitro studies of PDT on different cell lines using various photosensitizers. El-Khatib and coworker [30] examined the effect of PDT with 5-ALA in primary meningioma cell lines. For PDT, about 5000 cells per well were plated in 20 wells of a blank 96-well plate. In each block of four wells, 150 μL of 0, 25, 50, and 100 μg/mL 5-ALA solutions was inoculated and one block was used as the negative control without 5-ALA and without light application. PDT was performed for 3 h using a laser (635 nm, 18.75 J/cm2). A cell viability assay was performed 90 min after PDT. The authors observed a significant and dose-dependent decrease of viability. Either 5-ALA or PDT alone did not affect viability [30]. Mirzaei and coworker [31] evaluated the photodynamic effect with radachlorin as photosensitizer on human liver cancer cells (HepG2) and normal liver cells (HFLF-PI4) measuring the viability using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) tetrazolium (MTT) assay. The photosensitizer radachlorin without light irradiation had no toxic effect on the cell lines. Cell survival of HepG2 and HFLF-PI4 cells were decreased following PDT in a concentration-dependent manner. The study group could also observe that the HepG2 cells were more sensitive to radachlorin PDT than HFLF-PI4 cells. The 50% lethal dose of radachlorin HepG2 cells were 30 μg/mL and 20 μg/mL, 24 h after exposure to doses of 5 J/cm2 and 15, or 25 J/cm2. To kill HepG2 cells with minimal effects on normal HFLF-PI4 cells the optimal radachlorin and light dose were 100 μg/mL and 15 J/cm2 [31]. Another study group investigated the potential use of 5-ALA PDT induced protoporphyrin IX (PPIX) on a nasopharyngeal carcinoma cell line. The cells were irradiated at 4 h after incubation with 5-ALA (10–200 μg/mL) by a diode laser (630 nm) at various energy levels (1–50 J/cm2). After incubation with 5-ALA, a time- and dose-dependent increase of cellular PPIX-fluorescence was seen up to a threshold concentration of 1000 μg/mL 5-ALA. The combination of 5-ALA and laser irradiation leaded to a significant, concentration-, energy-, and time-dependent increase of cell death. Cellular survival at 100 μg/mL ALA and 10 J/cm2 laser irradiation was <5% after 48 h [32]. Guan and coauthors [33] evaluated PDT with MB on an osteosarcoma-derived cell line (UMR 106). The photocytotoxicity on the cell line was investigated 24 h after MB PDT using sulforhodamine B assay (SRB) and light microscopy. MB under red light irradiation caused a drug-concentration (0–100 μM) and light-dose (0–32 J/cm2) dependent cytotoxicity. Furthermore, the SRB assay and light microscopy showed a significant decrease of cell numbers after LED light-activated MB treatment (100 μM, 32 J/cm2) [33]. It is particularly interesting to investigate the effect of PDT in therapy resistant tumor cells in HNSCC. For this purpose, two cell lines have been included in this study. SCC-25 cells were originally isolated from the primary tumor of a patient with tongue carcinoma [34,35]. SCC-25 cells formed tumors in severe combined immunodeficiency (SCID) mice but not in athymic nude mice suggesting less aggressive behavior. Moreover, SCC-25 induced tumors did not develop regional or distant metastasis in mouse models [36]. In vitro, SCC-25 cells were found to be radioresistant [37]. SCC-25 cells contain a deletion and a frame shift in the TP53 gene protein coding region and do not synthesize any p53 protein (own Sanger sequencing results). This cell line represents a typical issue also common in HNSCC patients, the p53 protein loss. Detroit 562 cells grow tumors and develop regional and lung metastases when injected in nude mice [38]. Detroit 562 was isolated from the malignant pleural effusion of a patient with pharyngeal carcinoma [39,40]. A frequent gain of function mutation R175H of TP53 gene is contained in Detroit 562 cells [41], which has been confirmed by us utilizing Sanger sequencing. Both SCC-25 and Detroit 562 cells were HPV-negative [41].PDF Image | Photodynamic Low Level Laser Squamous Cell Carcinoma
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