Theodoro Viviane, de Oliveira Fujii Lucas, Lucke Leticia Dudri, Bortolazzo Fernanda Oriani, Silva Daniela Fernanda Dezotti, Carneiro Giane Daniela, do Amaral Maria Esméria Corezola, de Oliveira Camila Andréa, de Andrade Thiago Antonio Moretti, Bombeiro André Luis, Vicente Cristina Pontes, do Bomfim Fernando Russo Costa, de Oliveira Alexandre Leite Rodrigues, Bagnato Vanderlei Salvador, Esquisatto Marcelo Augusto Marretto, Mendonça Fernanda Aparecida Sampaio, Dos Santos Gláucia Maria Tech, de Aro Andrea Aparecida
Biomedical Sciences Graduate Program, University Center of Herminio Ometto Foundation / FHO, Araras, São Paulo, Brazil.
Department of Structural and Functional Biology, Institute of Biology, State University of Campinas - UNICAMP, Campinas, São Paulo, Brazil.
Heliyon. 2020 May 12;6(5):e03882. doi: 10.1016/j.heliyon.2020.e03882. eCollection 2020 May.
The objective of this study was to evaluate the effects of red Light Emiting Diode (red LED) irradiation on fibroblasts in adipose-derived mesenchymal stem cells (ASC) co-culture on the scratch assay. We hypothesized that red LED irradiation could stimulate paracrine secretion of ASC, contributing to the activation of genes and molecules involved in cell migration and tissue repair. ASC were co-cultured with NIH/3T3 fibroblasts through direct contact and subjected to red LED irradiation (1.45 J/cm/5min6s) after the scratch assay, during 4 days. Four groups were established: fibroblasts (F), fibroblasts + LED (FL), fibroblasts + ASC (FC) and fibroblasts + LED + ASC (FLC). The analyzes were based on and expression, quantification of collagen types I and III, tenomodulin, VEGF, TGF-β1, MMP-2 and MMP-9, as well as viability analysis and cell migration. Higher expression was observed in FC compared to F. Group FC presented higher amount of tenomodulin and VEGF in relation to the other groups. In the cell migration analysis, a higher number of cells was observed in the scratched area of the FC group on the 4 day. There were no differences between groups considering cell viability, expression, amount of collagen types I and III, MMP-2 and TGF-β1, whereas TGF-β1 was not detected in the FC group and the MMP-9 in none of the groups. Our hypothesis was not supported by the results because the red LED irradiation decreased the healing response of ASC. An inhibitory effect of the LED irradiation associated with ASC co-culture was observed with reduction of the amount of TGF-β1, VEGF and tenomodulin, possibly involved in the reduced cell migration. In turn, the ASC alone seem to have modulated fibroblast behavior by increasing , VEGF and tenomodulin, leading to greater cell migration. In conclusion, red LED and ASC therapy can have independent effects on fibroblast wound healing, but the combination of both does not have a synergistic effect. Therefore, future studies with other parameters of red LED associated with ASC should be tested aiming clinical application for tissue repair.
本研究的目的是评估红光发光二极管（红色LED）照射对脂肪来源间充质干细胞（ASC）共培养的成纤维细胞在划痕试验中的影响。我们假设红色LED照射可以刺激ASC的旁分泌分泌，促进参与细胞迁移和组织修复的基因和分子的激活。通过直接接触将ASC与NIH/3T3成纤维细胞共培养，并在划痕试验后进行红色LED照射（1.45 J/cm/5min6s），持续4天。建立了四组：成纤维细胞（F）、成纤维细胞+LED（FL）、成纤维细胞+ASC（FC）和成纤维细胞+LED+ASC（FLC）。分析基于 和 表达、I型和III型胶原蛋白、腱调蛋白、VEGF、TGF-β1、MMP-2和MMP-9的定量，以及活力分析和细胞迁移。与F组相比，FC组观察到更高的 表达。与其他组相比，FC组的腱调蛋白和VEGF含量更高。在细胞迁移分析中，第4天在FC组的划痕区域观察到更多的细胞。在细胞活力、 表达、I型和III型胶原蛋白、MMP-2和TGF-β1的含量方面，各组之间没有差异，而在FC组中未检测到TGF-β1，在任何组中均未检测到MMP-9。我们的假设未得到结果支持，因为红色LED照射降低了ASC的愈合反应。观察到与ASC共培养相关的LED照射具有抑制作用，TGF-β1、VEGF和腱调蛋白的量减少，这可能与细胞迁移减少有关。反过来，单独的ASC似乎通过增加 、VEGF和腱调蛋白来调节成纤维细胞的行为，并导致更大的细胞迁移。总之，红色LED和ASC疗法对成纤维细胞伤口愈合可能具有独立作用，但两者的组合没有协同作用。因此，未来应测试与ASC相关的红色LED的其他参数的研究，以用于组织修复的临床应用。
