不同强度持续静态压力对兔腰椎小关节软骨细胞活性影响的实验研究  

Effects of continuous hydrostatic pressures on chondrocytes of the lumbar facet .joint in rabbits

作　　者：郑帅[1] 王吉兴[1] 任海龙[1] 张继业[1] 姜欢畅[1] 

机构地区：[1]南方医科大学南方医院脊柱骨科,广州510515

出　　处：《中华创伤骨科杂志》2012年第3期232-235,共4页Chinese Journal of Orthopaedic Trauma

基　　金：广东省自然科学基金（9151051501000027）

摘　　要：目的探讨不同强度持续静态压力对体外培养的兔腰椎小关节软骨细胞活性的影响。方法分离新西兰白兔腰椎小关节软骨细胞，体外培养。将培养的第3代软骨细胞分为4组：对照组、30kPa组、60kPa组和90kPa持续静态加压组。显微镜下观察其形态，分别用免疫组化、噻唑蓝（MTT）、酶联免疫吸附法（ELISA）法鉴定软骨细胞，检测细胞增殖及Ⅱ型胶原的合成情况。结果第3代软骨细胞Ⅱ型胶原免疫组织化学染色，显示为阳性。软骨细胞增殖OD值，1d除了对照组与30kPa组相比差异无统计学意义外，其他各组两两比较差异均有统计学义义（P〈0．05）；2～4d：除了30kPa组与60kPa绀相比差异无统计学意义外，其他各组两两比较差异均有统计学意义（P〈0．05）；5～10d：各组间两两比较差异均有统计学意义（P〈0．05）。空白对照组的Ⅱ型胶原含精最高[（7．517±0．328）μg／L]，其次是60kPa组[（6．035±0．075）μg／L]，90kPa组含量最低[（2．873±0．127）μg／L]，30kPa组的Ⅱ型胶原含量为（4．846±0．093）μg／L，各组间两两比较差异均有统计学意义（P〈0．05）。结论兔腰椎小关节软骨细胞在持续静态压力下容易发生退变，细胞增殖率和Ⅱ型胶原含量均下降。Objective To investigate the effects of different continuous hydrostatic pressures on the chondrocytes of the lumbar facet jnint cuhured in vitro in rabbits. Methods The cartilage veils of the lumbar factor joint in New Zealand white rabbits were isolated and cultured in vitro, lmmunohistoehemistry was used to identify tire cell type. The third generation of the cartilage cells were put into a self-designed pressure chamber and subjected to different continuous hydrostatic pressures （0, 30, 60, and 90 kPa）. The morphological changes of the cells were observed in all groups by the I,aser Scanning Confocal Microscopy. 3-（4, 5-1）imethylthiazol-2-yl）-2, 5-diphenyltetrazolium bromide （MTT） was adopted to test the increment rate of tire cartilage cells. The cnzwne-linked immunosorbent assay （ELISA） was perIormed to measure contents of collagen II in the supernatant fluid of chondrocytes in different groups. Results lmmunohisiochemistry showed a positive staining of collagen II of the third generation of the cartilage cells. MTT showed significant differences（ P 〈 0. 05） between either 2 grnups except between 0 and 30 kPa gnmps on day one, except between 30 and 60 kPa groups on days 2 to 4, and with no exception on days 5 to 10. EI,ISA showed that the 0 kPa group had tire highest content of collagen II （7. 517 ± 0. 328 μg/L）, followed by the 60 kPa group （6. 035 ±0. 075 μg/L）, the 30 kPa group （4. 846 ±0. 093 μg/L） and the 90 kPa group （2. 873 ±0. 127 μg/L）, with significant between-group differences （ P 〈0. 05）. Conclusion A continuous hydrostatic pressure, especially an excessively low or high one, can result in a significant decrease in proliteration of the chondrocytes of the lumbar facet joint cultured in vitro in rabbits and in collagen II content as well which may induce degeneration of the chondrocytes.

关 键 词：腰椎 关节 软骨细胞 压力 胶味Ⅱ型 

分 类 号：R681.3[医药卫生—骨科学]