Abstract
The current study evaluated the photobiomodulatory effect of visible red light on cell proliferation and viability in various fibroblast diabetic models in vitro, namely, unstressed normal (N) and stressed normal wounded (NW), diabetic wounded (DW), hypoxic wounded (HW) and diabetic hypoxic wounded (DHW). Cells were irradiated at a wavelength of 660 nm with a fluence of 5 J/cm2 (11.23 mW/cm2), which related to an irradiation time of 7 min and 25 s. Control cells were not irradiated (0 J/cm2). Cells were incubated for 48 h and cellular proliferation was determined by measuring 5-bromo-2′-deoxyuridine (BrdU) in the S-phase (flow cytometry), while viability was assessed by the Trypan blue exclusion test and Apoptox-glo triplex assay. In comparison with the respective controls, PBM increased viability in N- (P ≤ 0.001), HW- (P ≤ 0.01) and DHW-cells (P ≤ 0.05). HW-cells showed a significant progression in the S-phase (P ≤ 0.05). Also, there was a decrease in the G2M phase in HW- and DHW-cells (P ≤ 0.05 and P ≤ 0.05, respectively). This study concludes that hypoxic wounded and diabetic hypoxic wounded models responded positively to PBM, and PBM does not damage stressed cells but has a stimulatory effect on cell viability and proliferation to promote repair and wound healing. This suggests that the more stressed the cells are the better they responded to photobiomodulation (PBM).
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References
Diabetes atlas (2015) http://www.idf.org/diabetesatlas. Accessed 22 Aug
Global status report on non-communicable diseases 2010 (2016) http://www.who.int/chp/ncd_global_status_report/en. Accessed 29 Aug
Guo S, DiPietro LA (2010) Critical review in oral biology & medicine: factors affecting wound healing. J Dent Res 89:219–229
Denadai AS, Aydos DR, Silva SL, Olmedo L, de Senna Cardoso BMS, da Silva AKB, Carvalho PTC (2015) Acute effects of low-level laser therapy (660 nm) on oxidative stress levels in diabetic rats with skin wounds. J Exp Ther Oncol 11:85–89
Chen Y, Yang S, Hu J, Yu C, He M, Cai Z (2016) Increased expression of SETD7 promotes cell proliferation by regulating cell cycle and indicates poor prognosis in hepatocellular carcinoma. PLoS One 11:e0154939
Francis-Goforth KN, Harken AH, Saba JD (2010) Normalization of diabetic wound healing. Surgery 147:446–449
Peppa M, Stavroulakis P, Raptis SA (2009) Advanced glycoxidation products and impaired diabetic wound healing. Wound Repair Regen 17:461–472
Wagener FA, Carels C, Lundvig D (2013) Targeting the redox balance in inflammatory skin conditions. Int J Mol Sci 14:9126–9167
Anders JJ, Lanzafame RJ, Arany PR (2015) Low-level light/laser therapy versus photobiomodulation therapy. Photomed Laser Surg 33:183–184. https://doi.org/10.1089/pho.2015.9848
Chaves MEA, de Araújo AR, ACC P, Pinotti M (2014) Effects of low-power light therapy on wound healing: LASER x LED*. An Bras Dermatol 89(4):616–623
Karu T (1999) Primary and secondary mechanisms of action of visible to near-IR radiation on cells. J Photochem Photobiol 49:1–17. https://doi.org/10.1016/S1011-1344(98)00219-X
Silveira PC, Silva LA, Pinho CA, Souza PS, Ronsani MM, Scheffer DL, Pinho RA (2013) Effects of low-level laser therapy (GaAs) in an animal model of muscular damage induced by trauma. Lasers Med Sci 28:431–436
Soares DM, Ginani F, Henriques A, Barboza C (2015) Effects of laser therapy on the proliferation of human periodontal ligament stem cells. Lasers Med Sci 30:1171–1174
Fujihara NA, Hiraki KR, Marque MM (2006) Irradiation at 780 nm increases proliferation rate of osteoblasts independently of dexamethasone presence. Lasers Surg Med 38:332–336
Tschon M, Incerti-Parenti S, Cepollaro S, Checchi L, Fini M (2015) Photobiomodulation with low-level diode laser promotes osteoblast migration in an in vitro micro wound model. J Biomed Opt 20:78002. https://doi.org/10.1117/1.JBO.20.7.078002
Stadler I, Evans R, Kolb B, Naim JO, Narayan V, Buehner N, Lanzafame RJ (2000) In vitro effects of low-level laser irradiation at 660 nm on peripheral blood lymphocytes. Lasers Surg Med 27:255–261
Eduardo FP, Mehnert DU, Monezi TA, Zezell DM, Schubert MM, Eduardo CP, Marques MM (2007) Cultured epithelial cells response to phototherapy with low-intensity laser. Lasers Surg Med 39:365–372. https://doi.org/10.1002/lsm.20481
Ejiri K, Aoki A, Yamaguchi Y, Ohshima M, Izumi Y (2014) High-frequency low-level diode laser irradiation promotes proliferation and migration of primary cultured human gingival epithelial cells. Lasers Med Sci 29:1339–1347. https://doi.org/10.1007/s10103-013-1292-7
Moore P, Ridgway TD, Higbee RG, Howard EW, Lucroy MD (2005) Effect of wavelength on low-intensity laser irradiation-stimulated cell proliferation in vitro. Lasers Surg Med 36:8–12. https://doi.org/10.1002/lsm.20117
Barboza CA, Ginani F, Soares DM, Henriques AC, Freitas Rde A (2014) Low-level laser irradiation induces in vitro proliferation of mesenchymal stem cells. Einstein (Sao Paulo) 12:75–81
Ginani F, Soares DM, Barreto MP, Barboza CA (2015) Effect of low-level laser therapy on mesenchymal stem cell proliferation: a systematic review. Lasers Med Sci 30:2189–2194. https://doi.org/10.1007/s10103-015-1730-9
YH W, Wang J, Gong DX, HY G, SS H, Zhang H (2012) Effects of low-level laser irradiation on mesenchymal stem cell proliferation: a microarray analysis. Lasers Med Sci 27:509–519. https://doi.org/10.1007/s10103-011-0995-x
Ayuk SM, Houreld NN, Abrahamse H (2012) Collagen production in diabetic wounded fibroblasts in response to low intensity laser irradiation (LILI) at 660 nm. Diabetes Technol Ther 14:1110–1117
Basso FG, Pansani TN, Turrioni APS, Bagnato VS, Hebling J, CAde S (2012) In vitro wound healing improvement by low-level laser therapy application in cultured gingival fibroblasts. Int J Dent 2012:719452
Hawkins D, Abrahamse H (2006) Effect of multiple exposures of low-level laser therapy on the cellular responses of wounded human skin fibroblasts. Photomed Laser Surg 24:705–714
Duan X, Ponomareva L, Veeranki S, Choubey D (2011) IFI16 induction by glucose restriction in human fibroblasts contributes to autophagy through activation of the ATM/AMPK/p53 pathway. PLoS One 6(5):e19532. https://doi.org/10.1371/journal.pone.0019532
Hemsley CM, Luo JX, Andreae CA, Butler CS, Soyer OS, Titball RW (2014) Bacterial drug tolerance under clinical conditions is governed by anaerobic adaptation but not anaerobic respiration. Antimicrob Agents Chemother 58(10):5775–5783
Houreld N, Abrahamse H (2007) Irradiation with a 632.8 nm helium-neon laser with 5 J/cm2 stimulates proliferation and expression of Interleukin-6 in diabetic wounded fibroblast cells. Diabetes Technol Ther 9(5):451–459
Rigau J, Sun C, Trelles MA, Berns M (1996) Effects of the 633 nm laser on the behaviour and morphology of primary fibroblasts in culture. Proc SPIE 3198:38–45
Liang CC, Park AY, Guan JL (2007) In vitro scratch assay: a convenient and inexpensive method for analysis of cell migration in vitro. Nat Protoc 2:329–333
Elosta A, Ghous T, Ahmed N (2012) Natural products as anti-glycation agents: possible therapeutic potential for diabetic complications. Curr Diabetes Rev 8(2):92–108
Tang Y, Chen A (2014) Curcumin eliminates the effect of advanced glycation end-products (AGEs) on the divergent regulation of gene expression of receptors of AGEs by interrupting leptin signalling. Lab Investig 94:503–516. https://doi.org/10.1038/labinvest.2014.42
Muller-Buhl U, Leutgeb R, Bungartz J, Szecsenyi J, Laux G (2013) Expenditure of chronic venous leg ulcer management in German primary care: results from a population-based study. Int Wound J 10:52–56. https://doi.org/10.1111/j.1742-481X.2012.00942.x
Rüttermann M, Maier-Hasselmann A, Nink-Grebe B, Burckhardt M (2013) Local treatment of chronic wounds in patients with peripheral vascular disease, chronic venous insufficiency, and diabetes. Dtsch Arztebl Int 110:25–31. https://doi.org/10.3238/arztebl.2013.0025.
Sekhejane PR, Houreld NN, Abrahamse H (2011) Irradiation at 636 nm positively affects diabetic wounded and hypoxic cells in vitro. Photomed Laser Surg 29:521–530
Alghamdi KM, Kumar A, Mouss NA (2012) Low-level laser therapy: a useful technique for enhancing the proliferation of various cultured cells. Lasers Med Sci 27:237–249. https://doi.org/10.1007/s10103-011-0885-2
Fekrazad R, Asefi S, Allahdadi M, Kalhori KA (2016) Effect of photobiomodulation on mesenchymal stem cells. Photomed Laser Surg 34:543–549. https://doi.org/10.1089/pho.2015.4028.
Al-Watban FAH (2009) Laser therapy converts diabetic wound healing to normal healing. Photomed Laser Surg 27(1):127–135
Houreld N, Abrahamse H (2010) Low-intensity laser irradiation stimulates wound healing in diabetic wounded fibroblast cells (WS1). Diabetes Technol Ther 12(12):971–978
Hamblin MR (2008) Mechanisms of low level light therapy, http://photobiology.info/Hamblin.html
Pellicioli AC, Martins MD, Dillenburg CS, Marques MM, Squarize CH, Castilho RM (2014) Laser phototherapy accelerates oral keratinocyte migration through the modulation of the mammalian target of rapamycin signaling pathway. J Biomed Opt 19:028002. https://doi.org/10.1117/1.JBO.19.2.028002
Houreld N, Masha R, Abrahamse H (2012) Low-intensity laser irradiation at 660 nm stimulates cytochrome c oxidase in stressed fibroblast cells. Lasers Surg Med 44:429–434. https://doi.org/10.1002/lsm.22027
Peplow P, Chung T, Baxter G (2010) Laser photobiomodulation of proliferation of cells in culture: a review of human and animal studies. Photomed Laser Surg 28(supplement 1):S3–S40
Woodruff L, Bounkeo J, Brannon W, Dawes K, Barham C, Waddell D, Enwemeka C (2004) The efficacy of laser therapy in wound repair: a meta-analysis of the literature. Photomed Laser Surg 22:241–247
Zungu IL, Hawkins Evans D, Abrahamse H (2009) Mitochondrial responses of normal and injured human skin fibroblasts following low level laser irradiation—an in vitro study. Photochem Photobiol 85:987–996. https://doi.org/10.1111/j.1751-1097.2008.00523.x
Pereira L, Longo J, Azevedo R (2012) Laser irradiation did not increase the proliferation or the differentiation of stem cells from normal and inflamed dental pulp. Arch Oral Biol 57:1079–1085. https://doi.org/10.1016/j.archoralbio.2012.02.012
Lamers M, Almeida M, Vicente-Manzanares M, Horwitz A, Santos M (2011) High glucose-mediated oxidative stress impairs cell migration. PLoS One 6:e22865. https://doi.org/10.1371/journal.pone.0022865
Schartinger V, Galvan O, Riechelmann H, Dudás J (2012) Differential responses of fibroblasts, non-neoplastic epithelial cells, and oral carcinoma cells to low-level lasertherapy. Support Care Cancer 20:523–529. https://doi.org/10.1007/s00520-011-1113-0
Häkkinen L, Uitto V, Larjava H (2000) Cell biology of gingival wound healing. Periodontology 24:127–152
Posten W, Wrone D, Dover J, Arndt K, Silapunt S, Alam M (2005) Low-level laser therapy for wound healing: mechanism and efficacy. Dermatol Surg 31:334–340
Wiegand C, Hipler U (2008) Methods for the measurement of cell and tissue compatibility including tissue regeneration process. GMS Krankenhhyg Interdiszip 3:1–9
Evans DH, Abrahamse H (2008) Efficacy of three different laser wavelengths for in vitro wound healing. Photodermatol, Photoimmunol & Photomed 24:199–210
Acknowledgements
All lasers were supplied and set up by the CSIR National Laser Centre.
Funding
This study was funded by the South African Research Chairs Initiative of the Department of Science and Technology and National Research Foundation of South Africa (Grant No. 98337), as well as grants received from the University of Johannesburg, the African Laser Centre (student bursary) and CSIR National Laser Centre Laser Rental Pool Program.
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This study received ethical approval from the University of Johannesburg, Faculty of Health Sciences Academic Ethics Committee (AEC01-13-2014).
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Ayuk, S.M., Houreld, N.N. & Abrahamse, H. Effect of 660 nm visible red light on cell proliferation and viability in diabetic models in vitro under stressed conditions. Lasers Med Sci 33, 1085–1093 (2018). https://doi.org/10.1007/s10103-017-2432-2
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DOI: https://doi.org/10.1007/s10103-017-2432-2