光生物调节作用对高糖环境下成骨细胞影响的体外实验研究

Effect of photobiomodulationon osteoblasts in high glucose environment in vitro

目的:研究高糖环境下光生物调节作用对成骨细胞增殖、分化的影响。方法:将成骨前体细胞MC3T3-E1分为对照组、光照组、高糖组和高糖+光照组,首先用CCK-8法检测各组24h、48h和72h的细胞活力,然后检测14天碱性磷酸酶染色情况及其活性,并对21天钙结节进行茜素红染色并半定量分析,实时荧光定量PCR检测21天成骨相关基因OCN、Runx2的表达,最后统计学分析各组间的差异。结果:与对照组相比,光照组在24h、48h、72h的细胞活力增强,14天时碱性磷酸酶染色及活性增强,21天钙结节形成增加、OCN和Runx2的表达上升;与对照组相比,高糖组在48h、72h的细胞活力下降,14天时碱性磷酸酶染色及活性减弱,21天钙结节形成减少、OCN及Runx2的表达降低;高糖+光照组与高糖组相比,在24h、48h、72h的细胞活力升高,14天时碱性磷酸酶染色及活性增强,21天钙结节形成增加、OCN和Runx2的表达增强;高糖+光照组与对照组相比,在21天时钙结节形成增加,其余指标差异无统计学意义。结论:在高糖环境下,成骨细胞经光生物调节作用,可补偿高糖对细胞增殖分化的抑制,可为促进高糖环境下颌骨缺损的修复提供理论依据。

Objective:To study the effect of photobiomodulation(PBM)on the proliferation and differentiation of osteoblasts in high glucose environment.Methods:MC3T3-E1 cells were divided into four groups:control group,light group high glucose group,and high glucose+light group.CCK-8 method was used to detect the cell viability of MC3T3-E1 cells at 24 h,48 h and 72 h,alkaline phosphatase staining and its activity at 14 d were detected,alizarin red staining and semi quantitative analysis were performed on calcium nodules at 21 d,and the expression of osteogenic related genes OCN and Runx2 at 21 d were detected by real-time quantitative PCR.Finally,the differences between the groups were statistically analyzed.Results:Compared with the control group,the cell viability of the light group increased at 24h,48h and 72h,the alkaline phosphatase staining and its activity increased at 14 d,the calcium nodule formation increased at 21 d,and the expression of OCN and Runx2 increased;Compared with the control group,the cell viability of high glucose group decreased at 48h and 72h,the staining and activity of alkaline phosphatase decreased at 14 d,the formation of calcium nodule decreased at 21 d,and the expression of OCN and Runx2 decreased;Compared with the high glucose group,the viability of cells in the high glucose+light group was increased at 24h,48h and 72h,the alkaline phosphatase staining and activity increased at 14 d,the calcium nodule formation increased at 21 d,and the expression of OCN and Runx2 increased;The calcium nodule formation increased at 21 d in the high glucose+light group and the other indexes were not statistically significant compared with the control group.Conclusion:Low level laser irradiation of osteoblasts can compensate the inhibition of cell proliferation and differentiation caused by high glucose,which can provide a theoretical basis for promoting the repair of jaw defects in high glucose environment.