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Erschienen in: Lasers in Medical Science 2/2018

20.11.2017 | Original Article

The effect of low-level laser therapy (660 nm) on the gene expression involved in tissue repair

verfasst von: Stheace Kelly Fernandes Szezerbaty, Rodrigo Franco de Oliveira, Deise A. A. Pires-Oliveira, Cristina Pacheco Soares, Daniele Sartori, Regina Célia Poli-Frederico

Erschienen in: Lasers in Medical Science | Ausgabe 2/2018

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Abstract

The effect of low-level laser therapy (LLLT) on the healing of skin lesions has been evaluated in many studies; however, the molecular mechanisms involved in the biostimulatory effects resulting from this treatment need to be better understood. The paper aims to analyze the effects of LLLT (660 nm) at doses of 1 and 5 J/cm2 on cell viability and expression of vascular endothelial growth factor (VEGF) and interleukin (IL6) genes in L929 fibroblast cells. The dose-response curve was performed with the GaInAlAs (660 nm) laser-treated cells at energy rates of 1 and 5 J/cm2. Cell viability was quantified at 24, 48, and 72 h after irradiation and the effects of TLBP on the cytoskeleton and endoplasmic reticulum were evaluated by fluorescence microscopy and the RT-qPCR method was used for the analysis of gene expression. It was observed that the 72 h group had a statistically significant increase in cell viability compared to the 48 h group (p < 0.01) and when compared to the 72 h control (p = 0.03). In 72 h, a greater distribution of the cytoskeleton filaments and the more evident endoplasmatic reticulum was verified, indicating an increase in the protein synthesis when compared with the control group. In the expression of the VEGF gene, a significant increase of 1.98 times (p < 0.05) in the number of transcripts was observed; whereas for the IL6 gene, a decrease of the transcripts was 4.05 times (p < 0.05), both occurring within 72 h after irradiation at 5 J/cm2. The LLLT (660 nm) at the dose of 5 J/cm2 should modulate cellular viability, upregulated VEGF, and downregulated IL6 expression of messenger RNA in culture of L929 fibroblast cells.
Literatur
1.
Zurück zum Zitat Henriques ACG, Cazal C, Castro JFL (2010) Ação da laserterapia no processo de proliferação e diferenciação celular: revisão da literatura. Rev Col Bras Cir 37(4):295–302CrossRefPubMed Henriques ACG, Cazal C, Castro JFL (2010) Ação da laserterapia no processo de proliferação e diferenciação celular: revisão da literatura. Rev Col Bras Cir 37(4):295–302CrossRefPubMed
2.
Zurück zum Zitat Santuzzi CH, Pedrosa DF, Nogueira BV, Buss HF, Freire MOVM, Gonçalves WLS (2011) Combined use of low level laser therapy and cyclooxygenase-2 selective inhibition on skin incisional wound reepithelialization in mice: a preclinical study. An Bras Dermatol 86(2):278–283CrossRefPubMed Santuzzi CH, Pedrosa DF, Nogueira BV, Buss HF, Freire MOVM, Gonçalves WLS (2011) Combined use of low level laser therapy and cyclooxygenase-2 selective inhibition on skin incisional wound reepithelialization in mice: a preclinical study. An Bras Dermatol 86(2):278–283CrossRefPubMed
3.
Zurück zum Zitat Barboza CAG, Ginani F, Soares DM, Henriques ACG, Freitas RA (2014) Low-level laser irradiation induces in vitro proliferation of mesenchymal stem cells. Einstein 12(1):75–81CrossRefPubMedPubMedCentral Barboza CAG, Ginani F, Soares DM, Henriques ACG, Freitas RA (2014) Low-level laser irradiation induces in vitro proliferation of mesenchymal stem cells. Einstein 12(1):75–81CrossRefPubMedPubMedCentral
4.
Zurück zum Zitat Karu T, Pyatibrat L, Kalendo G (1995) Irradiation with He-Ne laser increases ATP level in cells cultivated in vitro. J Photochem Photobiol B27(3):219–222CrossRef Karu T, Pyatibrat L, Kalendo G (1995) Irradiation with He-Ne laser increases ATP level in cells cultivated in vitro. J Photochem Photobiol B27(3):219–222CrossRef
5.
Zurück zum Zitat Werner S, Grose R (2003) Regulation of wound healing by growth factors and cytokines. Physiol Rev 83:835–870CrossRefPubMed Werner S, Grose R (2003) Regulation of wound healing by growth factors and cytokines. Physiol Rev 83:835–870CrossRefPubMed
6.
Zurück zum Zitat Lins RDAU, Lucena KCR, Granville-Garcia AF, Dantas EM, Catão MHCV, Neto LGC (2010) Biostimulation effects of low-power laser in the repair process. An Bras Dermatol 85(6):849–855CrossRefPubMed Lins RDAU, Lucena KCR, Granville-Garcia AF, Dantas EM, Catão MHCV, Neto LGC (2010) Biostimulation effects of low-power laser in the repair process. An Bras Dermatol 85(6):849–855CrossRefPubMed
7.
Zurück zum Zitat Kajdaniuk D, Marek B, Borgiel-Marek H, Kos-Kudla B (2011) Vascular endothelial growth factor (VEGF)-part 1: in physiology and pathophysiology. Endokrynol Pol 62:444–455PubMed Kajdaniuk D, Marek B, Borgiel-Marek H, Kos-Kudla B (2011) Vascular endothelial growth factor (VEGF)-part 1: in physiology and pathophysiology. Endokrynol Pol 62:444–455PubMed
8.
Zurück zum Zitat Guirro ECO, Montebelo MIL, Bortot BA, Torres MACB, Polacow ML (2010) Effect of laser (670 nm) on healing of wounds covered with occlusive dressing: a histologic and biomechanicalanalysis. Photomed Laser Surg 28(5):629–634CrossRef Guirro ECO, Montebelo MIL, Bortot BA, Torres MACB, Polacow ML (2010) Effect of laser (670 nm) on healing of wounds covered with occlusive dressing: a histologic and biomechanicalanalysis. Photomed Laser Surg 28(5):629–634CrossRef
9.
Zurück zum Zitat Szymanska J, Goralczyk K, Klawe JJ, Lukowicz M, Michalska M, Goralczyk B, Zalewski P, Newton JL, Gryko L, Zajac A, Rosc D (2013) Phototherapy with low-level laser influences the proliferation of endothelial cells and vascular endothelial growth factor and transforming growth factor-beta secretion. J Physiol Pharmacol 64(3):387–391PubMed Szymanska J, Goralczyk K, Klawe JJ, Lukowicz M, Michalska M, Goralczyk B, Zalewski P, Newton JL, Gryko L, Zajac A, Rosc D (2013) Phototherapy with low-level laser influences the proliferation of endothelial cells and vascular endothelial growth factor and transforming growth factor-beta secretion. J Physiol Pharmacol 64(3):387–391PubMed
10.
Zurück zum Zitat Meireles GC, Santos JN, Chagas PO, Moura AP, Pinheiro AL (2008) Effectiveness of laser photobiomodulation at 660 or 780 nanometers on the repair of third-degree burns in diabetic rats. Photomed Laser Surg 26:47–54CrossRefPubMed Meireles GC, Santos JN, Chagas PO, Moura AP, Pinheiro AL (2008) Effectiveness of laser photobiomodulation at 660 or 780 nanometers on the repair of third-degree burns in diabetic rats. Photomed Laser Surg 26:47–54CrossRefPubMed
11.
Zurück zum Zitat Chiarotto GB, Neves LMG, Esquisatto MAM, Amaral MEC, Santos GMT, Mendonça FAZ (2014) Effects of laser irradiation (670-nm InGaPamd 830-nm GaAlAs) on burn os second-degree in rats. Lasers Med Sci 29:1685–1693CrossRefPubMed Chiarotto GB, Neves LMG, Esquisatto MAM, Amaral MEC, Santos GMT, Mendonça FAZ (2014) Effects of laser irradiation (670-nm InGaPamd 830-nm GaAlAs) on burn os second-degree in rats. Lasers Med Sci 29:1685–1693CrossRefPubMed
12.
Zurück zum Zitat Mosmann T (1983) Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 65(1–2):55–63CrossRefPubMed Mosmann T (1983) Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 65(1–2):55–63CrossRefPubMed
13.
Zurück zum Zitat Pfaffl MW (2001) A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Res 29(9):2003–2007CrossRef Pfaffl MW (2001) A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Res 29(9):2003–2007CrossRef
14.
Zurück zum Zitat Araújo CE, Ribeiro MS, Favaro R, Zezell DM, Zorn TM (2007) Ultrastructural and autoradiographical analysis show a faster skin repair in He-Ne laser-treated wounds. J Photochem Photobiol B 86(2):87–96CrossRefPubMed Araújo CE, Ribeiro MS, Favaro R, Zezell DM, Zorn TM (2007) Ultrastructural and autoradiographical analysis show a faster skin repair in He-Ne laser-treated wounds. J Photochem Photobiol B 86(2):87–96CrossRefPubMed
15.
Zurück zum Zitat de Eduardo P, Bueno DF, de Freitas PM, Marques MM, Passos-Bueno MR, Eduardo Cde P, Zatz M (2008) Stem cell proliferation under low intensity laser irradiation: a preliminary study. Lasers Surg Med 40(6):433–438CrossRef de Eduardo P, Bueno DF, de Freitas PM, Marques MM, Passos-Bueno MR, Eduardo Cde P, Zatz M (2008) Stem cell proliferation under low intensity laser irradiation: a preliminary study. Lasers Surg Med 40(6):433–438CrossRef
16.
Zurück zum Zitat Marques JM, Pacheco-Soares C, Da Silva NS (2014) Evaluation of the photobiomodulation in L929 cell culture. Exp Biol Med (Maywood) 239(12):1638–1643CrossRef Marques JM, Pacheco-Soares C, Da Silva NS (2014) Evaluation of the photobiomodulation in L929 cell culture. Exp Biol Med (Maywood) 239(12):1638–1643CrossRef
17.
Zurück zum Zitat Zanotti GB, Oliveira PI, Reis SFS, Silva FS, Araújo AR (2011) Efeitos do laser de baixa potência sobre a regeneração da cartilagem na osteoartrose. Rev Fisio Bras 12(2):139–146 Zanotti GB, Oliveira PI, Reis SFS, Silva FS, Araújo AR (2011) Efeitos do laser de baixa potência sobre a regeneração da cartilagem na osteoartrose. Rev Fisio Bras 12(2):139–146
18.
Zurück zum Zitat Maiya AG, Kumar P, Nayak S (2009) Photo-stimulatory effect of low energy helium-neon laser irradiation on excisional diabetic wound healing dynamics in wistar rats. Indian J Dermatol 54(4):323–329CrossRefPubMedPubMedCentral Maiya AG, Kumar P, Nayak S (2009) Photo-stimulatory effect of low energy helium-neon laser irradiation on excisional diabetic wound healing dynamics in wistar rats. Indian J Dermatol 54(4):323–329CrossRefPubMedPubMedCentral
19.
Zurück zum Zitat Rocha Júnior AM, Vieira BJ, Andrade LCF, Aarestrup FM (2007) Effects of low-level laser therapy on the progress of wound healing in humans: the contribution of in vitro and vivo experimental studies. J Vasc Bras 6(3):257–265CrossRef Rocha Júnior AM, Vieira BJ, Andrade LCF, Aarestrup FM (2007) Effects of low-level laser therapy on the progress of wound healing in humans: the contribution of in vitro and vivo experimental studies. J Vasc Bras 6(3):257–265CrossRef
20.
Zurück zum Zitat Carnevalli CMM, Pacheco-Soares C, Zângaro RA, Pinheiro ALB, Silva NS (2003) Laser light prevents apoptosis on CHO K-1 cell line. J Clin Laser Med Surg 21:193–196CrossRefPubMed Carnevalli CMM, Pacheco-Soares C, Zângaro RA, Pinheiro ALB, Silva NS (2003) Laser light prevents apoptosis on CHO K-1 cell line. J Clin Laser Med Surg 21:193–196CrossRefPubMed
21.
Zurück zum Zitat Cury V, Moretti AIS, Assis L, Bossini P, Crusca JS, BenattiNeto C, Fangel R, Souza HP, Hamblin MR, Parizotto NA (2013) Low level laser therapy increases angiogenesis in a model of ischemic skin flap in rats mediated by VEGF, HIF-1a and MMP-2. J Photochem Photobiol B 125:164–170CrossRefPubMedPubMedCentral Cury V, Moretti AIS, Assis L, Bossini P, Crusca JS, BenattiNeto C, Fangel R, Souza HP, Hamblin MR, Parizotto NA (2013) Low level laser therapy increases angiogenesis in a model of ischemic skin flap in rats mediated by VEGF, HIF-1a and MMP-2. J Photochem Photobiol B 125:164–170CrossRefPubMedPubMedCentral
22.
Zurück zum Zitat Feng J, Zhang Y, Xing D (2012) Low-power laser irradiation (LPLI) promotes VEGF expression and vascular endothelial cell proliferation through the activation of ERK/Sp1 pathway. Cell Signal 24:1116–1125CrossRefPubMed Feng J, Zhang Y, Xing D (2012) Low-power laser irradiation (LPLI) promotes VEGF expression and vascular endothelial cell proliferation through the activation of ERK/Sp1 pathway. Cell Signal 24:1116–1125CrossRefPubMed
23.
Zurück zum Zitat Houreld NN, Ayuk SM, Abrahamnse H (2014) Expression of genes in normal fibroblast cells (WS1) in response to irradiation at 660 nm. J Photochem Photobiol B130:146–152 Houreld NN, Ayuk SM, Abrahamnse H (2014) Expression of genes in normal fibroblast cells (WS1) in response to irradiation at 660 nm. J Photochem Photobiol B130:146–152
24.
Zurück zum Zitat Albertini R, Villaverde AB, Aimbire F, Bjordal J, Brugnera A, Mittmann J, Silva JA, Costa M (2008) Cytokine mRNA expression is decreased in the subplantar muscle of rat paw subjected to carrageenan-induced inflammation after low-level laser therapy. Photomed Laser Surg 26(1):19–24CrossRefPubMed Albertini R, Villaverde AB, Aimbire F, Bjordal J, Brugnera A, Mittmann J, Silva JA, Costa M (2008) Cytokine mRNA expression is decreased in the subplantar muscle of rat paw subjected to carrageenan-induced inflammation after low-level laser therapy. Photomed Laser Surg 26(1):19–24CrossRefPubMed
25.
Zurück zum Zitat Boschi ES, Leite CE, Saciura VC, Caberlon E, Lunardelli A, Bitencourt S, Melo DA, Oliveira JR (2008) Anti-inflammatory effects of low-level laser therapy (660 nm) in the early phase in carrageenan-induced pleurisy in rat. Lasers Surg Med 40(7):500–508CrossRefPubMed Boschi ES, Leite CE, Saciura VC, Caberlon E, Lunardelli A, Bitencourt S, Melo DA, Oliveira JR (2008) Anti-inflammatory effects of low-level laser therapy (660 nm) in the early phase in carrageenan-induced pleurisy in rat. Lasers Surg Med 40(7):500–508CrossRefPubMed
26.
Zurück zum Zitat Francisco G, Hernández C, Simó R (2006) Serum markers of vascular inflammation in dyslipidemia. Clin Chim Acta 369:1–16CrossRefPubMed Francisco G, Hernández C, Simó R (2006) Serum markers of vascular inflammation in dyslipidemia. Clin Chim Acta 369:1–16CrossRefPubMed
Metadaten
Titel
The effect of low-level laser therapy (660 nm) on the gene expression involved in tissue repair
verfasst von
Stheace Kelly Fernandes Szezerbaty
Rodrigo Franco de Oliveira
Deise A. A. Pires-Oliveira
Cristina Pacheco Soares
Daniele Sartori
Regina Célia Poli-Frederico
Publikationsdatum
20.11.2017
Verlag
Springer London
Erschienen in
Lasers in Medical Science / Ausgabe 2/2018
Print ISSN: 0268-8921
Elektronische ISSN: 1435-604X
DOI
https://doi.org/10.1007/s10103-017-2375-7

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