Wu Jinjie, Wu Yuehao, Chen Xuening, Zhi Wei
National Engineering Research Center for Biomaterials, Sichuan University, Chengdu 610041, P.R.China.
Key Laboratory of Advanced Technologies of Materials (Ministry of Education), School of Materials Science and Engineering, Southwest Jiaotong University, Chengdu 610031, P.R.China.
Sheng Wu Yi Xue Gong Cheng Xue Za Zhi. 2020 Feb 25;37(1):96-104. doi: 10.7507/1001-5515.201903056.
This study investigated the early mechanical adaptability and osteogenic differentiation of mouse bone marrow mesenchymal stem cells (M-BMSCs) under micro-vibration stimulation (MVS). M-BMSCs were stimulated by MVS , cell proliferation, alkaline phosphatase (ALP) activity assay, and cytoskeleton were measured, and cell apoptosis was observed by flow cytometry. Early osteoblast-associated genes, runt-related transcription factor 2 (Runx2), Collagen Ⅰ (Col-Ⅰ) and ALP, were observed by RT-PCR and the activation of extracellular regulated protein kinases 1/2 (ERK1/2) was determined by Western blotting. The results showed that MVS had no significant effect on the proliferation of M-BMSCs. The early apoptosis was induced by mechanical stimulation (for one day), but the apoptosis was decreased after cyclic stimulation for 3 days. At the same time, MVS significantly accelerated the expression of F-actin protein in cytoskeleton, the synthesis of ALP and the ERK1/2 pathway, also up-regulated the expressions of Runx2, Col-Ⅰ and ALP genes. This study indicates that MVS could regulate cellular activity, alter early adaptive structure and finally promote the early osteogenic differentiation of M-BMSCs.
本研究调查了小鼠骨髓间充质干细胞(M-BMSCs)在微振动刺激(MVS)下的早期力学适应性和成骨分化。用MVS刺激M-BMSCs,检测细胞增殖、碱性磷酸酶(ALP)活性及细胞骨架,并通过流式细胞术观察细胞凋亡。采用RT-PCR检测早期成骨细胞相关基因,即 runt 相关转录因子2(Runx2)、Ⅰ型胶原蛋白(Col-Ⅰ)和ALP,并通过蛋白质免疫印迹法测定细胞外调节蛋白激酶1/2(ERK1/2)的激活情况。结果显示,MVS对M-BMSCs的增殖无显著影响。机械刺激(持续1天)诱导早期细胞凋亡,但在循环刺激3天后细胞凋亡减少。同时,MVS显著加速了细胞骨架中F-肌动蛋白蛋白的表达、ALP的合成及ERK1/2信号通路,还上调了Runx2、Col-Ⅰ和ALP基因的表达。本研究表明,MVS可调节细胞活性,改变早期适应性结构,最终促进M-BMSCs的早期成骨分化。