周期性牵张力对成骨细胞细胞骨架影响的实验研究

Effects of mechanical strain on osteoblast cytoskeleton of rat by DioDynamic testing system

目的 对大鼠成骨细胞施加单轴向周期性牵张应力,观察单轴向周期性牵张应力对大鼠成骨细胞细胞骨架的影响.方法 无菌环境下,组织分离法培养SD大鼠颅骨成骨细胞,体外培养,应用DioDynamic细胞应力加载系统分别对2组第3代大鼠成骨细胞加载0%和2%的单轴向周期性牵张应力,使用免疫荧光法检测细胞骨架蛋白表达,使用激光共聚焦显微镜观察细胞骨架的变化,随即抽取视野并测量各组细胞骨架蛋白表达的荧光强度,进行统计学分析.结果 经统计学计算,施加单轴向周期性牵张应力实验组与空白对照组相比,实验组大鼠成骨细胞骨架蛋白的荧光表达减弱;应力纤维变细,明显沿受力方向分布.结论 DioDynamic细胞应力加载系统可以很好地模拟生物体内成骨细胞所受单向周期牵张应力;大鼠成骨细胞细胞骨架感知单轴向周期性牵张应力,参与细胞的力学信号转导和细胞形态重构.

Objective To investigate the effects of mechanical strain on osteoblast cytoskeleton of rat by DioDynamic Testing System. Methods Osteoblasts were retrieved from SD rats＇ skull in the sterilized environment. DioDynamic testing system was used to form 0% and 2% mechanical strain on two groups of the third generation rats＇ osteoblasts. Immunofluorescent was used to detect expression of F-actin in the two groups of osteoblasts. Leica Microsystem software was applied to evaluate the fluorescence of F-actin expressed among the two groups of samples. Results Significant decrease was observed on the expression of protein F-actin in the 2%mechanical strain experinental group compared to the control group （0% mechanical strain）. Conclusion The DioDynamic testing system can successfully simulate the mechanical strain in vitro. Osteoblast cytoskeleton of rat participates in mechanical cell signal transduction and cell morphological reconstruction by detecting the mechanical strain.