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International Journal of Molecular Sciences Article Photodynamic Effect of Methylene Blue and Low Level Laser Radiation in Head and Neck Squamous Cell Carcinoma Cell Lines Barbara Kofler *, Angela Romani, Christian Pritz ID , Teresa Bernadette Steinbichler, Volker Hans Schartinger, Herbert Riechelmann and Jozsef Dudas ID Department of Otorhinolaryngology, Medical University of Innsbruck, 6020 Innsbruck, Austria; angela.romani@tirol-kliniken.at (A.R.); christianpritz@gmail.com (C.P.); teresa.steinbichler@tirol-kliniken.at (T.B.S.); volker.schartinger@i-med.ac.at (V.H.S.); herbert.riechelmann@i-med.ac.at (H.R.); jozsef.dudas@i-med.ac.at (J.D.) * Correspondence:ba.kofler@tirol-kliniken.at;Tel.:+43-50-504-23141 Received: 22 February 2018; Accepted: 4 April 2018; Published: 7 April 2018 Abstract: Photodynamic therapy (PDT) is suggested to have an impact on the treatment of early stage head and neck cancers (HNSCC). We investigated the effect of PDT with methylene blue (MB) and a diode laser (660 nm) as the laser source on HNSCC cell lines as an in vitro model of surface oral squamous cell carcinoma. Cell-cultures were exposed to 160 μM MB for 4 min and to laser light for 8 min. Viability was proven via cell viability assay and clonogenic survival via clone counting assay. The combination of MB and diode laser evidenced high efficient loss of cell viability by 5% of the control, while treatment with the same concentration of MB for 4 min alone showed a viability of 46% of the control. In both SCC-25 and Detroit 562 HNSCC cells, MB combined with the laser allowed a significant abrogation of clonogenic growth (p < 0.01), especially in the case of Detroit 562 cells less than 1% of the suspension plated cells were able to grow tumor cell nests. Multiresistant (Detroit 562) HNSCC cells expressing cancer stem cell markers are sensitive to MB/red laser combined PDT. Keywords: photodynamic therapy; methylene blue; head and neck carcinoma; TissueFaxs 1. Introduction Head and neck malignancies compose about 90% of squamous cell carcinoma (SCC). Depending on several factors including stage of cancer and condition of the patient, different treatment options are available. Main treatment principles include surgery, radiotherapy, chemotherapy, immune therapy, or combinations thereof [1–3]. Occasionally, other treatment modalities may be beneficial in selected patients. Photodynamic therapy (PDT) is a treatment modality for precancerous skin lesions with good cosmetic results and few adverse effects and allows application to multiple lesions [4]. Several studies reported successful treatment of early stage head and neck squamous cell carcinoma (HNSCC) and its precursor lesions with photodynamic therapy (PDT) [5,6]. Recently, an overall response rate of 91% with a complete response rate of 71% in early stage oral cavity and oropharyngeal cancer was observed in 170 HNSCC-patients. In these studies, the photosensitizer meta-tetrahydroxyphenylchlorin (mTHPC) was combined with a diode laser at 652 nm for PDT [7]. Rigual and coworker [8] assessed the response of dysplasia, intraepithelial neoplasia, and T1 carcinomas of the oral cavity to PDT with porfimer sodium and light at 630 nm. Responses were evaluated at 1 week, 1 month, and 3 months following intervention. Initially, 25 patients had a complete response, one a partial response, and one patient did not respond. Three patients with oral dysplasia with an initial complete response had a recurrence in the treatment Int. J. Mol. Sci. 2018, 19, 1107; doi:10.3390/ijms19041107 www.mdpi.com/journal/ijmsPDF Image | Photodynamic Low Level Laser Squamous Cell Carcinoma
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