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Int. J. Mol. Sci. 2018, 19, 1107 13 of 17 5. Conclusions MB was used in relatively high concentrations for short time treatments followed by 8 min of 660 nm laser exposure in this study, as an experimental model for PDT for easily accessible oral SCC. MB alone caused a significant toxicity, but in the preferred conditions of 4 min 160 μM MB and 8 min laser exposure 41% of cell viability reduction was light specific. This combination was also particularly effective against cancer stem cells, which were enriched in the multiresistant Detroit 562 cell culture. MB, as we also observed, is relative unselective, for better targeting several research groups try to pack it into nanoparticles as published by Usacheva and coauthors [48]. MB as a PDT agent for the treatment of respiratory tract cancer in animal models was described to be efficient [71]. Further studies are needed to assess the safety and efficacy of MB-associated PDT for the treatment of cancer in humans. Acknowledgments: Authors are grateful for the support of Heltschl Medicine Technique, Schlüsslberg, Austria. Parts of the study were supported by the Austrian Science Funds (FWF) Project Number: P25869-B13. Role of the Funding Sources: the funding sources had no influence on the direction and the outcome of the study. Author Contributions: Jozsef Dudas, Herbert Riechelmann, Teresa Bernadette Steinbichler and Volker Hans Schartinger conceived and designed the experiments; Jozsef Dudas prepared cell pellet embedding and paraffin sections, and performed immunohistochemical analysis, Angela Romani and Jozsef Dudas performed the cell treatments and cell-based experiments; Jozsef Dudas performed the TissueFaxs scanning and Tissuequest quantification, Christian Pritz developed and performed the clonogenic survival quantifications; Barbara Kofler and Herbert Riechelmann statistically analyzed the data; Angela Romani contributed reagents/materials/analysis tools; Barbara Kofler wrote the paper and all authors revised and improved the manuscript to its final form. Authorship was limited to those who have contributed substantially to the work reported. Conflicts of Interest: The authors declare no conflict of interest. Abbreviations 5-ALA 5-aminolevulinic acid hydrochloride APDT antimicrobial photodynamic therapy DAPI diamidino-2-phenylindole DMEM/F-12 Dulbecco’s Modified Eagle Medium/Nutrient Mixture F-12 EDTA ethylenediaminetetraacetic acid FISH fluorescence in situ hybridization FBS fetal bovine serum HNSCC head and neck squamous cell carcinoma HPPH 2-(1-Hexyloxyethyl)-2-devinyl pyropheophorbide HPV Human papilloma virus LLL low level laser MB methylene blue mTHPC meta-tetrahydroxyphenylchlorin MTT 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide PBS Phosphate Buffered Saline PDT Photodynamic therapy PPIX protoporphyrin IX SCC squamous cell carcinoma SCID severe combined immunodeficiency SRB sulforhodamine B assay References 1. Sunar, U. Monitoring photodynamic therapy of head and neck malignancies with optical spectroscopies. World J. Clin. Cases 2013, 1, 96–105. [CrossRef] [PubMed] 2. Hopper, C.; Niziol, C.; Sidhu, M. The cost-effectiveness of Foscan mediated photodynamic therapy (Foscan-PDT) compared with extensive palliative surgery and palliative chemotherapy for patients with advanced head and neck cancer in the UK. Oral Oncol. 2004, 40, 372–382. [CrossRef] [PubMed]PDF Image | Photodynamic Low Level Laser Squamous Cell Carcinoma
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