间歇性机械拉伸对体外培养兔关节软骨细胞微管蛋白-β取向的影响
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国家自然科学基金重点资助(10332060)


Effect of intermittent tension on the cell proliferation and β-tubu-lin reorientation of rabbit articular chondrocytes in vitro
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    摘要:

    目的探讨间歇性机械拉伸对兔关节软骨细胞生长和细胞形态及骨架的影响,为软骨细胞内力学信号传导研究的提供参考。方法采用FX-4000TM柔性基底拉伸系统对体外单层原代培养的兔软骨细胞实施正弦波、0.5Hz、05%间歇性周期应变加载,每日拉伸3h,共加载3天。对照组静态培养。3天后检测细胞活力、基质合成情况以及微管蛋白-β细胞免疫荧光染色,并对细胞周期进行检测。结果加载组软骨细胞生长良好,氨基多聚糖和蛋白多糖的合成与对照组无明显差异;但细胞增殖指数显著增高(P<0.05);微管蛋白-β沿细胞长轴发生重排,细胞形态和取向趋于一致。结论间歇性机械拉伸对细胞生长起到积极的作用,并且通过微管蛋白-β的重排影响细胞形态和取向。

    Abstract:

    Objective To investigate the relationship between cell proliferation and β-tubulin of rabbit articular chondrocytes under intermittent tension. Methods Chondrocytes were cultured in monolayer on flexible substrates and subjected to cyclic strain 0-5% with sinusoidal waveforms at 0.5 Hz for 3h/ day over 3days by using FX-4000TTM unit. Cell viability, extracelluar matrix, β-tubulin immunofluorescence and cell cycle were measured respectively. Results The cell viability and extracelluar matrix tests showed that articular chondrocytes cultured in monolayer grew well in vitro. There was no difference in the extracellular matrix synthesis between the loaded group and control group. However, the cell proliferation index increased significantly under intermittent tension. Furthermore, cultured chondrocytes underwent significant morphological changes when subjected to intermittent tensile stress. The cells elongated and aligned in a certain direction. Accompanying this shape change was a reorganization of β-tubulin in the direction of the cells’ long axis. Conclusion Our findings suggest that intermittent tension may have an active effect on the cell growth, and β-tubulin of articular chondrocytes plays an important role in the transduction of intermittent tensile stress which cause reorientation of articular chondrocytes.

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李晓娜,陈静,董江峰,陈维毅,吴文周.间歇性机械拉伸对体外培养兔关节软骨细胞微管蛋白-β取向的影响[J].医用生物力学,2007,22(4):373-377

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