低能量激光通过激活ROS/ERK通路促进张力诱导下MC3T3-E1成骨分化的机制研究OA

Objective This study aimed to investigate the effect of Low-Level Laser Therapy(LLLT)on the function of stress-induced osteogenic differentiation of mouse Embryonic Osteoblasts Cells(MC3T3-E1)and to study the mechanism of action in depth.Methods We divided MC3T3-E1 into control group(CON,no special treatment),stress group(Stress group,subjected to force treatment),laser group(LLLT group,8 J/cm² energy irradiation,irradiated once every other day,each irradiation for 20 s,irradiated for a total of 3 times),and combined group(Stress+LLLT group,laser treatment was performed 2 h after tension treatment).The osteogenic indexes of the cells in each group were detected by alkaline phosphatase(ALP)and Alizarin Red S(ARS)staining.The expression levels of Runt-related transcription factor 2(Runx2),ALP,and osteocalcin(OCN)were detected by qRT-PCR and western blotting.Reactive oxygen species(ROS)and western blotting were utilized to detect changes in the extracellular regulated protein kinases(ERK)pathway.Results The results of ALP and ARS staining showed that both Stress and LLLT treatments alone significantly increased the expression of ALP and ARS,while the combined treatment of the two had a more significant effect.Expression of osteogenesis-related genes and proteins showed that both Stress and LLLT treatment alone increased the expression levels of Runx2 and OCN.In contrast,the combined treatment had a more significant effect.The fluorescence staining and phosphorylation level of ERK showed that both Stress and LLLT treatments alone significantly increased the expression levels of ROS and p-ERK,while the combined treatment had a more significant effect.Conclusion This study concludes that LLLT can activate the ROS/ERK pathway,thereby promoting the osteogenic differentiation of MC3T3-E1 under tension induction.This finding provides an experimental basis for a deeper understanding of the mechanism of LLLT in osteoblastic activity,which has potential application value.