Laser Research Centre, University of Johannesburg, Johannesburg 2006, South Africa.
Cells. 2023 Apr 4;12(7):1080. doi: 10.3390/cells12071080.
Diabetic foot ulcers (DFUs) are open chronic wounds that affect diabetic patients due to hyperglycaemia. DFUs are known for their poor response to treatment and frequently require amputation, which may result in premature death. The present study evaluated the effect of photobiomodulation (PBM) at 660 nm on wound healing via activation of Ras/MAPK signalling in diabetic wounded cells in vitro. This study used four human skin fibroblast cell (WS1) models, namely normal (N), wounded (W), diabetic (D), and diabetic wounded (DW). Cells were irradiated at 660 nm with 5 J/cm. Non-irradiated cells (0 J/cm) served as controls. Cells were incubated for 24 and 48 h post-irradiation, and the effect of PBM on cellular morphology and migration rate, viability, and proliferation was assessed. Basic fibroblast growth factor (bFGF), its phosphorylated (activated) receptor FGFR, and phosphorylated target proteins (Ras, MEK1/2 and MAPK) were determined by enzyme-linked immunosorbent assay (ELISA) and Western blotting; nuclear translocation of p-MAPK was determined by immunofluorescence. PBM resulted in an increase in bFGF and a subsequent increase in FGFR activation. There was also an increase in downstream proteins, p-Ras, p-MEK1/2 and p-MAPK. PBM at 660 nm led to increased viability, proliferation, and migration as a result of increased bFGF and subsequent activation of the Ras/MAPK signalling pathway. Therefore, this study can conclude that PBM at 660 nm stimulates in vitro diabetic wound healing via the bFGF-activated Ras/MAPK pathway.
糖尿病足溃疡(DFUs)是由于高血糖影响糖尿病患者的开放性慢性伤口。DFUs 的治疗反应差,经常需要截肢,这可能导致过早死亡。本研究通过体外糖尿病创伤细胞中 Ras/MAPK 信号的激活,评估了 660nm 光生物调节(PBM)对伤口愈合的影响。本研究使用了四种人皮肤成纤维细胞(WS1)模型,分别为正常(N)、创伤(W)、糖尿病(D)和糖尿病创伤(DW)。细胞用 660nm 照射 5J/cm。未照射的细胞(0J/cm)作为对照。细胞在照射后 24 和 48 小时孵育,并评估 PBM 对细胞形态和迁移率、活力和增殖的影响。通过酶联免疫吸附试验(ELISA)和 Western blot 测定碱性成纤维细胞生长因子(bFGF)及其磷酸化(激活)受体 FGFR 以及磷酸化靶蛋白(Ras、MEK1/2 和 MAPK);通过免疫荧光测定核转位的 p-MAPK。PBM 导致 bFGF 增加,随后 FGFR 激活增加。下游蛋白 p-Ras、p-MEK1/2 和 p-MAPK 也增加。660nm 的 PBM 导致 bFGF 增加,随后 Ras/MAPK 信号通路激活,从而导致活力、增殖和迁移增加。因此,本研究可以得出结论,660nm 的 PBM 通过 bFGF 激活的 Ras/MAPK 通路刺激体外糖尿病伤口愈合。