短时间机械循环压力对3D培养脊柱终板软骨细胞的影响  

Short-term effects on the 3D cultured spinal endplate chondrocytes in vitro by cyclic mechanical stimulation

作　　者：张巍[1] 徐宏光[1] 俞云飞[1] 

机构地区：[1]皖南医学院附属弋矶山医院脊柱外科,安徽芜湖241001

出　　处：《皖南医学院学报》2015年第1期17-20,29,共5页Journal of Wannan Medical College

基　　金：国家自然科学基金项目(30973025;81311130314;81272048);安徽省自然科学基金项目(1308085MH152)

摘　　要：目的:探究短时间的机械循环压力对终板软骨细胞的影响,并探讨其机制。方法:大鼠终板软骨细胞的分离制备大鼠终板软骨细胞琼脂糖模板进行培养,然后通过FX-5000T对大鼠终板软骨细胞琼脂糖模板加载短时间的机械循环压力。通过活死染色对细胞进行活死分析,通过RT-PCR和Western blotting检测ANK基因表达。通过RT-PCR和ELISA检测TGF-β1的表达。结果:大鼠终板软骨细胞加载短时间的机械循环压力后ANK基因和TGF-β1表达量增加,加载短时间的机械循环压力的实验组跟实验对照组活死染色没有显著改变。结论:短时间的压力刺激下终板软骨ANK基因和TGF-β1表达量上调抑制终板软骨的退变,短时间的压力刺激并不影响终板软件细胞的活死。这个实验结果可能在椎间盘退变的防治中提供一种新的方法。Objective： To investigate the mechanisms and short-term impact on the 3D cultured spinal endplate chondrocytes in vitro by exerting cyclic mechanical stimulation. Methods： The endplate chondrocytes were isolated from rats and cultured in agarose constructs for 48 hours. Then FX-5000 TTM Flexercell Tension PlusTM unit was applied to exert short-term cyclic mechanical stimulation on the cultured chondrocytes. Endplate chondrocytes viability was determined with LIVE / DEAD viability / cytotoxicity kit,and ANK gene expression was examined by RT-PCR and Western blot. Both RT-PCR and ELISA were performed to measure TGF-β1 expression. Results： Short-term cyclic mechanical stimulation increased the ANK gene and TGF-β1 expression in rat endplate chondrocytes. Live / Dead assay verified that the non-loading（ NC） group and experimental group had no change in viability following the application of short-term cyclic mechanical compression. Conclusion： Short-term cyclic mechanical stimulation increased the ANK gene and TGF-β1 expression in endplate chondrocytes and inhibited the cartilage degeneration of endplate,yet failed to cause viability of the chondrocytes. The findings suggest that this may be a new approach to treatment of the intervertebral disc degeneration.

关 键 词：短时间机械循环压力 终板软骨细胞 椎间盘 ANK基因 TGF-Β1表达 

分 类 号：R681.5[医药卫生—骨科学]