低频振动对小鼠骨髓间充质干细胞成骨分化及相关基因表达的影响

Effects of low-frequency vibration on osteogenic differentiation of mouse bone marrow mesenchymal stem cells

目的观察低频振动对小鼠骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs)成骨分化的影响。方法取3周龄C57BL/6小鼠,分离及培养其BMSCs。将细胞分为两组,对照组及低频振动组,两组均诱导BMSCs进行成骨分化,低频振动组在诱导过程中实施低频振动干预(15 Hz,垂直加速度0.3 g),对照组不进行干预,分别于第7天、第14天收获细胞,将第7天的细胞进行碱性磷酸酶(alkaline phosphatase, ALP)染色,将第14天的细胞进行茜素红染色;并利用Real-time PCR检测相关基因表达。结果低频振动组细胞第7天的ALP活性明显高于对照组,且在14 d形成的钙结节多于对照组;所检测基因中,低频振动组细胞Ⅰ型胶原蛋白、骨桥蛋白、骨钙素、骨保护素基因的表达均高于对照组,而破骨细胞分化因子(receptor activator of NF-κB ligand, RANKL)较对照组低。结论低频振动可以促进BMSCs的成骨分化,并调节成骨细胞分泌的细胞因子,影响破骨细胞的生成。

Objective To observe the effect of low-frequency vibration on the osteoblastogenesis of mouse bone marrow mesenchymal stem cells(BMSCs). Methods Three-week old C57 BL/6 mouse was used to isolate and culture BMSCs. The cells were divided into the control group and low-frequency vibration treated group, and they were induced by the osteogenic induction medium. At the 7 th day and the 14 th day, the cells were harvested. Alkaline phosphatase(ALP) staining test was used at the 7 th day and alizarin red staining at the14 th day. The genes expression was detected by real-time PCR. Results The ALP staining positive area in lowfrequency group was increased at 7 th day as compared with the control group. The alizarin red staining showed that the formation of calcium nodules in low-frequency group was increased compared to the control group. The genes expression levels of collagen type I, osteopontin, osteocalcin, osteoprotegerin were increased in lowfrequency group compared to the control group. The gene expression of receptor activator of NF-κB ligand was decreased by low-frequency vibration. Conclusion The low-frequency vibration could promote the osteogenic differentiation of BMSCs and influence the osteoclastogenesis by secreting cytokine.