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Photobiomodulation is associated with a decrease in cell viability and migration in oral squamous cell carcinoma

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Abstract

The treatment of squamous cell carcinoma (SCC) involves surgery, chemotherapy, and/or radiotherapy, which can cause mucositis (inflammation of the oral mucosa that causes considerable pain and can compromise the continuity of oncological treatment). Photobiomodulation (PBM) has been successfully used in the treatment of mucositis, but doubts arise regarding the use of laser for areas in which tumor cells may remain. In this study, the effect of PBM on the viability, mitochondrial activity, proliferation, apoptosis, and migration of cells derived from oral SCC was evaluated. SCC9 cells were irradiated with laser (660 and 780 nm, using 11 dosimetric parameters) and submitted to mitochondrial and caspase 3 activity tests after 1 and 3 days. Based on the results, cell viability (neutral red assay), proliferation (BrdU assay), and migration (scratch-wound assay) were evaluated using only the dosimetric parameters recommended for mucositis. Non-irradiated cells served as the control. The experiments were performed in triplicate. The 11 parameters diminished mitochondrial activity and induced tumor cell apoptosis. Using the parameters recommended for mucositis, irradiation with 780 nm (70 mW, 4 J/cm2) proved to be the safest and led to a reduction in cell viability, the induction of apoptosis, and a reduction in the migration capacity of the tumor cells.

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References

  1. Marocchio LS, Lima J, Sperandio FF, Corrêa L, de Sousa SO (2010) Oral squamous cell carcinoma: an analysis of 1,564 cases showing advances in early detection. J Oral Sci 52(2):267–273

    Article  PubMed  Google Scholar 

  2. Sperandio FF, Giudice FS, Corrêa L, Pinto DS Jr, Hamblin MR, de Sousa SC (2013) Low-level laser therapy can produce increased aggressiveness of dysplastic and oral cancer cell lines by modulation of Akt/mTOR signaling pathway. J Biophotonics 6(10):839–847

    CAS  PubMed  PubMed Central  Google Scholar 

  3. Saba NF, Goodman M, Ward K, Flowers C, Ramalingam S, Owonikoko T, Chen A, Grist W, Wadsworth T, Beitler JJ, Khuri FR, Shin DM (2011) Gender and ethnic disparities in incidence and survival of squamous cell carcinoma of the oral tongue, base of tongue, and tonsils: a surveillance, epidemiology and end results program-based analysis. Oncology 81(1):12–20

    Article  PubMed  PubMed Central  Google Scholar 

  4. Antunes HS, Ferreira EM, de Matos VD, Pinheiro CT, Ferreira CG (2008) The impact of low power laser in the treatment of conditioning-induced oral mucositis: a report of 11 clinical cases and their review. Med Oral Patol Oral Cir Bucal 13(3):E189–E192

    PubMed  Google Scholar 

  5. Genot-Klastersky MT, Klastersky J, Awada F, Awada A, Crombez P, Martinez MD, Jaivenois MF, Delmelle M, Vogt G, Meuleman N, Paesmans M (2008) The use of low-energy laser (LEL) for the prevention of chemotherapy- and/or radiotherapy-induced oral mucositis in cancer patients: results from two prospective studies. Support Care Cancer 16(12):1381–1387

    Article  CAS  PubMed  Google Scholar 

  6. Simões A, Eduardo FP, Luiz AC, Campos L, Sá PH, Cristófaro M, Marques MM, Eduardo CP (2009) Laser phototherapy as topical prophylaxis against head and neck cancer radiotherapy-induced oral mucositis: comparison between low and high/low power lasers. Lasers Surg Med 41(4):264–270

    Article  PubMed  Google Scholar 

  7. Bensadoun RJ, Nair RG (2012) Low-level laser therapy in the prevention and treatment of cancer therapy-induced mucositis: 2012 state of the art based on literature review and meta-analysis. Curr Opin Oncol 24(4):363–370

    Article  CAS  PubMed  Google Scholar 

  8. Schartinger VH, Galvan O, Riechelmann H, Dudás J (2012) Differential responses of fibroblasts, non-neoplastic epithelial cells, and oral carcinoma cells to low-level laser therapy. Support Care Cancer 20(3):523–529

    Article  PubMed  Google Scholar 

  9. Zecha JA, Raber-Durlacher JE, Nair RG, Epstein JB, Sonis ST, Elad S, Hamblin MR, Barasch A, Migliorati CA, Milstein DM, Genot MT, Lansaat L, van der Brink R, Arnabat-Dominguez J, van der Molen L, Jacobi I, van Diessen J, de Lange J, Smeele LE, Schubert MM, Bensadoun RJ (2016) Low level laser therapy/photobiomodulation in the management of side effects of chemoradiation therapy in head and neck cancer: part 1: mechanisms of action, dosimetric, and safety considerations. Support Care Cancer 24(6):2781–2792

    Article  PubMed  PubMed Central  Google Scholar 

  10. Zecha JA, Raber-Durlacher JE, Nair RG, Epstein JB, Elad S, Hamblin MR, Barasch A, Migliorati CA, Milstein DM, Genot MT, Lansaat L, van der Brink R, Arnabat-Dominguez J, van der Molen L, Jacobi I, van Diessen J, de Lange J, Smeele LE, Schubert MM, Bensadoun RJ (2016) Low-level laser therapy/photobiomodulation in the management of side effects of chemoradiation therapy in head and neck cancer: part 2: proposed applications and treatment protocols. Support Care Cancer 24(6):2793–2805

    Article  PubMed  PubMed Central  Google Scholar 

  11. Sonis ST, Hashemi S, Epstein JB, Nair RG, Raber-Durlacher JE (2016) Could the biological robustness of low level laser therapy (Photobiomodulation) impact its use in the management of mucositis in head and neck cancer patients. Oral Oncol 54:7–14

    Article  PubMed  Google Scholar 

  12. Russo G, Haddad R, Posner M, Machtay M (2008) Radiation treatment breaks and ulcerative mucositis in head and neck cancer. Oncologist 13(8):886–898

    Article  PubMed  Google Scholar 

  13. Rosenthal DI, Trotti A (2009) Strategies for managing radiation-induced mucositis in head and neck cancer. Semin Radiat Oncol 19(1):29–34

    Article  PubMed  Google Scholar 

  14. Pinheiro AL, Carneiro NS, Vieira AL, Brugnera A Jr, Zanin FA, Barros RA, Silva PS (2002) Effects of low-level laser therapy on malignant cells: in vitro study. J Clin Laser Med Surg 20(1):23–26

    Article  PubMed  Google Scholar 

  15. Gomes Henriques AC, Ginani F, Oliveira RM, Keesen TS, Galvão Barboza CA, Oliveira Rocha HA, de Castro JF, Della Coletta R, de Almeida Freitas R (2014) Low-level laser therapy promotes proliferation and invasion of oral squamous cell carcinoma cells. Lasers Med Sci 29(4):1385–1395

    PubMed  Google Scholar 

  16. Hawkins D, Houreld N, Abrahamse H (2005) Low level laser therapy (lllt) as an effective therapeutic modality for delayed wound healing. Ann N Y Acad Sci 1056:486–493

    Article  CAS  PubMed  Google Scholar 

  17. Myakishev-Rempel M, Stadler I, Brondon P, Axe DR, Friedman M, Nardia FB, Lanzafame R (2012) A preliminary study of the safety of red light phototherapy of tissues harboring cancer. Photomed Laser Surg 30(9):551–558

    Article  PubMed  PubMed Central  Google Scholar 

  18. Gao X, Xing D (2009) Molecular mechanisms of cell proliferation induced by low power laser irradiation. J Biomed Sci 16:4

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  19. Huang YY, Chen ACH, Carroll JD, Hamblin MR (2009) Biphasic dose response in low level light therapy. Dose Response 7:358–383

    Article  PubMed  PubMed Central  Google Scholar 

  20. Huang YY, Sharma SK, Carroll J, Hamblin MR (2011) Biphasic dose response in low level light therapy - an update. Dose Response 9(4):602–618

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Chung H, Dai T, Sharma SK, Huang YY, Carroll JD, Hamblin MR (2012) The nuts and bolts of low-level laser (light) therapy. Ann Biomed Eng 40(2):516–533

    Article  PubMed  Google Scholar 

  22. AlGhamdi KM, Kumar A, Moussa NA (2012) Low-level laser therapy: a useful technique for enhancing the proliferation of various cultured cells. Lasers Med Sci 27(1):237–249

    Article  PubMed  Google Scholar 

  23. Dias Schalch T, Porta Santos Fernandes K, Costa-Rodrigues J, Pereira Garcia M, Agnelli Mesquita-Ferrari R, Kalil Bussadori S, Fernandes MH (2016) Photomodulation of the osteoclastogenic potential of oral squamous carcinoma cells. J Biophotonics 9(11–12):1136–1147

    Article  CAS  PubMed  Google Scholar 

  24. Nogueira GT, Mesquita-Ferrari RA, Souza NH, Artilheiro PP, Albertini R, Bussadori SK, Fernandes KP (2012) Effect of low-level laser therapy on proliferation, differentiation, and adhesion of steroid-treated osteoblastos. Lasers Med Sci 27:1189–1193

    Article  PubMed  Google Scholar 

  25. Fujihara NA, Hiraki KR, Marques MM (2006) Irradiation at 780 nm increases proliferation rate of osteoblasts independently of dexamethasone presence. Lasers Surg Med 38:332–336

    Article  PubMed  Google Scholar 

  26. Silva DF, Mesquita-Ferrari RA, Fernandes KP, Raele MP, Wetter NU, Deana AM (2012) Effective transmission of light for media culture, plates and tubes. Photochem Photobiol 88(5):1211–1216

    Article  CAS  PubMed  Google Scholar 

  27. Hughes SE (2003) Detection of apoptosis using in situ markers for DNA strand breaks in the failing human heart. Fact or epiphenomenon? J Pathol 201(2):181–186

    Article  PubMed  Google Scholar 

  28. Grivicich I, Regner A, da Rocha AB (2007) Morte Celular por Apoptose. Rev Bras Cancerol 53(3):335–343

    Google Scholar 

  29. Fotakis G, Timbrell JA (2006) In vitro cytotoxicity assays: comparison of LDH, neutral red, MTT and protein assay in hepatoma cell lines following exposure to cadmium chloride. Toxicol Lett 160(2):171

    Article  CAS  PubMed  Google Scholar 

  30. Chiba K, Kawakami K, Tohyama K (1998) Simultaneous evaluation of cell viability by neutral red, MTT and crystal violet staining assays of the same cells. Toxicol in Vitro 12(3):251–258

    Article  CAS  PubMed  Google Scholar 

  31. Van Tonder A, Joubert AM, Cromarty AD (2015) Limitations of the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay when compared to three commonly used cell enumeration assays. BMC Res Notes 8:47

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  32. Mossmann T (1983) Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 65:55–63

    Article  Google Scholar 

  33. Marshall NJ, Goodwin CJ, Holt SJ (1995) A critical assessment of the use of microculture tetrazolium assays to measure cell growth and function. Growth Regul 5(2):69–84

    CAS  PubMed  Google Scholar 

  34. Hamblin MR (2017) Mechanisms and applications of the anti-inflammatory effects of photobiomodulation. AIMS Biophys 4(3):337–361

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  35. Passarella S, Karu T (2014) Absorption of monochromatic and narrow band radiation in the visible and near IR by both mitochondrial and non-mitochondrial photoacceptors results in photobiomodulation. J Photochem Photobiol B 140:344–358

    Article  CAS  PubMed  Google Scholar 

  36. Wang S, Yu H, Wickliffe JK (2011) Limitation of the MTT and XTT assays for measuring cell viability due to superoxide formation induced by nano-scale TiO2. Toxicol in Vitro 25(8):2147–2151

    Article  CAS  PubMed  Google Scholar 

  37. Abrahamse H (2015) Stimulation of cellular proliferation and migration: is it a viable measure of photobiomodulation? Photomed Laser Surg 33(7):347–348

    Article  PubMed  Google Scholar 

  38. Morgan CD, Mills KC, Lefkowitz DL, Lefkowitz SS (1991) An improved colorimetric assay for tumor necrosis factor using WEHI 164 cells cultured on novel microtiter plates. J Immunol Methods 145(1–2):259–262

    Article  CAS  PubMed  Google Scholar 

  39. Iguchi H, Tanaka S, Ozawa Y, Kashiwakuma T, Kimura T, Hiraga T, Ozawa H, Kono A (1996) An experimental model of bone metastasis by human lung cancer cells: the role of parathyroid hormone-related protein in bone metastasis. Cancer Res 56:4040–4043

    CAS  PubMed  Google Scholar 

  40. Deyama Y, Tei K, Yoshimura Y, Izumiyama Y, Takeyama S, Halta M, Totsuka Y, Suzuki K (2008) Oral squamous cell carcinomas stimulate osteoclast differentiation. Oncol Rep 20(3):663–668

    CAS  PubMed  Google Scholar 

  41. Tang CH, Chuang JY, Fong YC, Maa MC, Way TD, Hung CH (2008) Bone-derived SDF-1 stimulates IL-6 release via CXCR4, ERK and NF-kappa B pathways and promotes osteoclastogenesis in human oral cancer cells. Carcinogenesis 29(8):1483–1492

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  42. Chuang FH, Hsue SS, Wu CW, Chen YK (2009) Immunohistochemical expression of RANKL, RANK, and OPG in human oral squamous cell carcinoma. J Oral Pathol Med 38(10):753–758

    Article  CAS  PubMed  Google Scholar 

  43. Tada T, Shin M, Fukushima H, Okabe K, Ozeki S, Okamoto M, Jimi E (2009) Oral squamous cell carcinoma cells modulate osteoclast function by RANKL-dependent and -independentmechanisms. Cancer Lett 274(1):126–131

    Article  CAS  PubMed  Google Scholar 

  44. Van Cann EM, Slootweg PJ, de Wilde PC, Otte-Höller I, Koole R, Stoelinga PJ, Merkx MA (2009) The prediction of mandibular invasion by squamous cell carcinomas with the expression of osteoclast-related cytokines in biopsy specimens. Int J Oral Maxillofac Surg 38(3):279–284

    Article  PubMed  Google Scholar 

  45. Jimi E, Furuta H, Matsuo K, Tominaga K, Takahashi T, Nakanishi O (2011) The cellular and molecular mechanisms of bone invasion by oral squamous cell carcinoma. Oral Dis 17(5):462–468

    Article  CAS  PubMed  Google Scholar 

  46. Honig A, Rieger L, Kapp M, Krockenberger M, Eck M, Dietl J, Kammerer U (2006) Increased tartrate resistant acid phosphatase (TRAP) expression in malignant breast, ovarian and melanoma tissue: an investigational study. BMC Cancer 6:199

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  47. How J, Brown JR, Saylor S, Rimm DL (2014) Macrophage expression of tartrate-resistant acid phosphatase as a prognostic indicator in colon cancer. Histochem Cell Biol 142(2):195–204

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  48. Vladimirov YA, Klebanov GI, Borisenko GG, Osipov AN (2004) Molecular and cellular mechanisms triggered by low-level laser irradiation. Biophysics 49:325–336

    Google Scholar 

  49. Kreslavski VD, Fomina IR, Los DA, Carpentier R, Kuznetsov VV, Allakhverdiev SI (2012) Red and near infra-red signaling: hypothesis and perspectives. J Photochem Photobiol C: Photochem Rev 13(3):190–203

    Article  CAS  Google Scholar 

  50. Bailes HJ, Lucas RJ (2013) Human melanopsin forms a pigment maximally sensitive to blue light (lambdamax approximately 479 nm) supporting activation of G(q/11) and G(i/o) signalling cascades. Proc Biol Sci 280:20122987

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  51. Hamblin MR (2018) Mechanisms and mitochondrial redox signaling in photobiomodulation. Photochem Photobiol 94(2):199–212

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  52. Fekrazad R, Chiniforush N (2014) Oral mucositis prevention and management by therapeutic laser in head and neck cancers. J Lasers Med Sci 5(1):1–7

    PubMed  PubMed Central  Google Scholar 

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Funding

This study was supported by São Paulo Research Foundation (grant no. 2013/07502-1). KPSF, SKB, and RAMF were supported by CNPq-National Council for Technological and Scientific Development (CNPq, grant nos. 311078/2015-0, 305905/2014-7, and 305739/2014-0).

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Authors and Affiliations

  1. Biophotonics Applied to Health Sciences Postgraduate Program, Nove de Julho University – UNINOVE, 235/249 Vergueiro Street, São Paulo, 01504-001, Brazil

    Tatiana Dias Schalch, Maria Fernanda Setúbal Destro Rodrigues, Mônica Pereira Garcia, Raquel Agnelli Mesquita Ferrari, Sandra Kalil Bussadori & Kristianne Porta Santos Fernandes

  2. Laboratory for Bone Metabolism and Regeneration; Faculdade de Medicina Dentária, Universidade do Porto, Dr. Manuel Pereira da Silva Street,, 4200-393, Porto, Portugal

    Maria Helena Fernandes, João Costa Rodrigues & Mônica Pereira Garcia

  3. Department of Oral Pathology, School of Dentistry, University of São Paulo, 2227 Professor Lineu Prestes Avenue,, São Paulo, 05508-000, Brazil

    Douglas Magno Guimarães & Fabio Daumas Nunes

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  1. Tatiana Dias Schalch
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  2. Maria Helena Fernandes
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  3. Maria Fernanda Setúbal Destro Rodrigues
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  4. Douglas Magno Guimarães
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  5. Fabio Daumas Nunes
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  6. João Costa Rodrigues
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  7. Mônica Pereira Garcia
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  8. Raquel Agnelli Mesquita Ferrari
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  9. Sandra Kalil Bussadori
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  10. Kristianne Porta Santos Fernandes
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Correspondence to Kristianne Porta Santos Fernandes.

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Schalch, T.D., Fernandes, M.H., Destro Rodrigues, M.F.S. et al. Photobiomodulation is associated with a decrease in cell viability and migration in oral squamous cell carcinoma. Lasers Med Sci 34, 629–636 (2019). https://doi.org/10.1007/s10103-018-2640-4

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