Abstract
The use of light emitting diodes (LED) as a therapeutic resource for wound healing has increased over the last years; however, little is still known about the molecular pathways associated to LED exposure. In the present study, we verified the effects of LED therapy on DNA methylation and expression of the DNA methyltransferase (Dnmt) genes, Dnmt1 and Dnmt3a, in an in vivo model of epithelial wound healing. Male Wistar rats were submitted to epithelial excision in the dorsal region and subsequently distributed within the experimental groups: group 1, animals that received irradiation of 0.8 J/cm2 of LED (604 nm); group 2, animals that received 1.6 J/cm2 of LED (604 nm); control (CTL), animals not submitted to therapeutic intervention. LED applications were performed during 7 days, and tissues from the periphery of the wound area were obtained for molecular analysis. The Image-J software was used for analysis of the wound area. DNA methylation was evaluated by ELISA-based method and gene expressions were quantified by real-time PCR. Decrease on global DNA methylation profile was observed in all experimental groups (CTL, 1, and 2) revealing the participation of DNA methylation in the healing process. Significant decrease in the wound area accompanied by increase in the Dnmt3a expression was associated to group 2. Based on our findings, we propose that DNA methylation is an important molecular mechanism associated to wound healing and that irradiation with 1.6 J/cm2 of LED evokes an increase in the expression of the Dnmt3a that might associates to the efficiency of the epithelial wound healing.
References
Mester E, Juhász J, Varga P, Karika G (1968) Lasers in clinical practice. Acta Chir Acad Sci Hung 9(3):349–57
Amid R, Kadkhodazadeh M, Ahsaie MG, Hakakzadeh A (2014) Effect of low level laser therapy on proliferation and differentiation of the cells contributing in bone regeneration. J Lasers Med Sci 5(4):163–70
Hadis MA, Zainal SA, Holder MJ, Carroll JD, Cooper PR, Milward MR, Palin WM. The dark art of light measurement: accurate radiometry for low-level light therapy. Lasers Med Sci 2016 Mar 10. [Epub ahead of print]
Safavi SM et al (2008) Effects of low-level He–Ne laser irradiation on the gene expression of IL-1beta, TNFalpha, IFN-gamma, TGF-beta, bFGF, and PDGF in rat’s gingiva. Lasers Med Sci 23(3):331–335
Chen CH, Tsai JL, Wang YH, Lee CL, Chen JK, Huang MH (2009) Low-level laser irradiation promotes cell proliferation and mRNA expression of type I collagen and decorin in porcine achilles tendon fibroblasts in vitro. J Orthop Res 27(5):646–50. doi:10.1002/jor.20800
Saygun I, Karacay S, Serdar M, Ural AU, Sencimen M, Kurtis B (2008) Effects of laser irradiation on the release of basic fibroblast growth factor (bFGF), insulin like growth factor-1 (IGF-1), and receptor of IGF-1 (IGFBP3) from gingival fibroblasts. Lasers Med Sci 23(2):211–15. doi:10.1007/s10103-007-0477-3
Chen CH, Hung HS, Hsu S (2008) Low energy laser irradiation increases endothelial cell proliferation, migration, and eNOS gene expression possibly via PI3K signal pathway. Lasers Surg Med 40(1):46–54. doi:10.1002/lsm.20589
Aleksic V et al (2010) Low-level Er: YAG laser irradiation enhances osteoblast proliferation through activation of MAPK/ERK. Lasers Med Sci 25(4):559–69. doi:10.1007/s10103-010-0761-5
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):237–49. doi:10.1007/s10103-011-0885-2
Anders JJ et al (2008) Light supports neurite outgrowth of human neural progenitor cells in vitro: the role of P2Y receptors. J Selected Top Quantum Electron 14(1):118–25. doi:10.1109/JSTQE.2008.916181
Dancáková L, Vasilenko T, Kováč I, Jakubčová K, Hollý M, Revajová V, Sabol F, Tomori Z, Iversen M, Gál P, Bjordal JM (2014) Low-level laser therapy with 810 nm wavelength improves skin wound healing in rats with Streptozotocin-induced diabetes. Photomed Laser Surg 32(4):198–204. doi:10.1089/pho.2013.3586
Karu TI (1987) Photobiological fundamentals of low-power therapy. J Quantum Electron 23(10):1703–17. doi:10.1590/S1413-35552009005000040
Nogueira VC, Coelho NP, Barros TL, Silva SM, Martins M, Arisawa EA (2014) Biomodulation effects of LED and therapeutic ultrasound combined with semipermeable dressing in the repair process of cutaneous lesions in rats. Acta Cir Bras 29(9):588–95. doi:10.1590/S0102-865020140015000
Wang G et al (2010) The effects of DNA methyltransferase inhibitors and histone deacetylase inhibitors on digit regeneration in mice. Regen Med 5:201–20. doi:10.2217/rme.09.91
Jones PA, Takai D (2001) The role of DNA methylation in mammalian epigenetics. Science 293(5532):1068–70. doi:10.1126/science.1063852
Jenuwein T, Allis CD (2001) Translating the histone code. Science 293(5532):1074–80. doi:10.1126/science.1063127
Vinck E et al (2003) Increased fibroblast proliferation induced by light emitting diode and low power laser irradiation. Lasers Med Sci 18(2):95–9. doi:10.1007/s10103-003-0262-x
Min PK, Goo BL (2013) 830 nm light-emitting diode low-level light therapy (LED-LLLT) enhances wound healing: a preliminary study. Laser Ther 22(1):43–49. doi:10.5978/islsm.13-OR-06
Adamskaya N et al (2011) Light therapy by blue LED improves wound healing in an excision model in rats. Injury 42(9):917–21. doi:10.1016/j.injury.2010.03.023
Gomes MV, Pelosi GG (2013) Epigenetic vulnerability and the environmental influence on health. Exp Biol Med (Maywood) 1(238):859–65. doi:10.1177/1535370213490630
Portela A, Esteller M (2010) Epigenetic modifications and human disease. Nat Biotechnol 28(10):1057–68. doi:10.1038/nbt.1685
Yanai H, Budovsky A, Tacutu R, Barzilay T, Abramovich A, Ziesche R, Fraifeld VE (2016) Tissue repair genes: the TiRe database and its implication for skin wound healing. Oncotarget Mar 31. doi: 10.18632/oncotarget.8501
Singh SM, Murphy B, O’Reilly RL (2003) Involvement of gene-diet/drug interaction in DNA methylation and its contribution to complex diseases: from cancer to schizophrenia. Clin Genet 64:451–460
Acknowledgments
This study was funded by the National Foundation for Development of Private Education (FUNADESP - Process N°5300859).
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that they have no conflict of interest.
Rights and permissions
About this article
Cite this article
Gomes, M.V.d.M., Manfredo, M.H., Toffoli, L.V. et al. Effects of the led therapy on the global DNA methylation and the expression of Dnmt1 and Dnmt3a genes in a rat model of skin wound healing. Lasers Med Sci 31, 1521–1526 (2016). https://doi.org/10.1007/s10103-016-2007-7
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10103-016-2007-7