力学刺激对体外立体培养软骨细胞基质代谢的影响  

Metabolize Analysis of Chondrocytes Cultured in Alginate in Response to Cyclical Dynamic Compression

作　　者：段王平[1] 苑伟[1] 孙振伟[1] 李琦[1] 赵昱[1] 卫小春[1] 

机构地区：[1]山西医科大学第二医院骨科,山西太原030001

出　　处：《实用骨科杂志》2015年第5期428-431,共4页Journal of Practical Orthopaedics

基　　金：国家自然科学基金项目(31340010;31271033);教育部高等学校博士学科点专项科研基金联合资助课题(20121417120004);山西省基础研究计划项目青年科技研究基金项目(2013021036-3);高等学校科技创新项目(20131105);山西医科大学博士启动基金项目(03201114)

摘　　要：目的分析周期性动态压缩刺激对体外海藻酸钠立体培养关节软骨细胞基质合成代谢的影响。方法取2月龄新西兰白兔10只,酶解消化获取膝关节全层软骨细胞,以海藻酸钠凝胶立体培养,分为实验组和对照组两组,对照组行静态培养,未施加任何压力;实验组利用Flexcell-5000力学加载系统对体外培养软骨细胞进行周期性压缩应力加载,1 h/d。于加载第7、14、21天留取软骨细胞,采用实时定量聚合酶联反应对软骨细胞蛋白聚糖(aggrecan,AGG)、Ⅱ型胶原、Ⅹ型胶原及基质金属蛋白酶-13(matrix metalloproteinase-13,MMP-13)mRNA进行定量分析。结果实验组在第7天AGG及Ⅱ型胶原mRNA的表达明显增高(P<0.05),随加载时间延长,其表达量逐渐下降,在第14、21天两组比较均无明显差异。同时,实验组在第7天时,Ⅹ型胶原及MMP-13的表达无明显差异。第14天,实验组Ⅹ型胶原及MMP-13 mRNA的表达与对照组比较明显增高(P<0.05)。结论立体培养软骨细胞在生理力学刺激7 d时可明显促进其基质合成能力,但随刺激时间的延长,其基质合成能力逐渐减弱,软骨细胞趋于肥大分化。Objective To analyze the matrix metabolite of isolated chondrocytes cultured in alginate in response to cyclical dynamic compression. Methods The chondrocytes were isolated from the 2-months rabbit knees cartilage, and the chondrocytes were cultured in alginate. The cells were treated with cyclical dynamic compression by using Flexcell - 5000 compression system kept under an uncompressed condition as a control. The mRNA gene expression of aggrecan （AGG） ,type Ⅱ collagen, type X collagen and matrix metalloproteinases 13 （MMP - 13 ） were determined by reverse transcription polymerase chain reaction （ RT - PCR） at 7,14 and 21 day after under loading. Results A significant up - regulation was observed in the gene expression of AGG and type Ⅱ collagen on 7 day in the experimental group compared to control group, but showed no significant difference on 14 and 21 days. The type X collagen and MMP - 13 mRNA gene expression were significantly increased on 14 days in the experimental group compared to control group. There were no significant difference on 7 and 21 days. Conclusion Chondrocytes cultured in alginate can keep their matrix synthetic ability under the physiological compression early, but the cells degenerate and hypertrophy over loading time.

关 键 词：软骨细胞 海藻酸钠 周期性动态压缩 代谢 

分 类 号：R318.01[医药卫生—生物医学工程]