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目的体外观察高强度周期性静水压对人膝关节软骨细胞形态和超微结构及其 II 型胶原分泌水平的影响。方法取正常成人膝关节软骨组织，采用两步酶消化法体外分离培养软骨细胞，分为2组：对照组，不施加压力；高压实验组，应用多功能恒温体外细胞培养中高压静水压力加载装置施加10.0 mPa 压力，共5天，每日2 h。甲苯胺蓝染色法鉴定软骨细胞；倒置相差显微镜观察加压后软骨细胞形态的变化；利用透射电镜观察加压后软骨细胞内超微结构的变化。II 型胶原免疫组织化学染色法检测并半定量分析加压后软骨细胞 II 型胶原分泌水平的变化。结果经高强度压力作用后，与对照组相比，高压实验组软骨细胞从形态上由多边形或不规则形变为长梭形，胞膜、胞质回缩，细胞量明显减少，生长稀疏；透射电镜观察显示，高压实验组软骨细胞出现染色体分布不均、染色体边集等细胞凋亡指征；II 型胶原免疫组织化学染色及半定量分析显示，高压实验组软骨细胞染色面积(43.76±5.61)%，较对照组染色面积(61.64±6.19)%相比，大小和程度均明显降低( P＜0.05)。结论超过人生理范围的高强度压力作用，可视为一种机械应力损伤，会引起软骨细胞形态和细胞内超微结构的改变，并导致软骨细胞的凋亡和蛋白分泌的下降，以上实验数据的获取，为进一步研究压力损伤与骨关节炎发病机制的联系奠定了实验基础。“,”Objective To investigate the effects of high-intensity cyclic hydrostatic pressures on the ultrastructure and type II collagen contents of chondrocytes of the knee joint cultured in vitro in human. Methods The normal chondrocytes of human knee joint were isolated and cultured in vitro. The 3rd generation of chondrocytes were treated with high-intensity cyclical hydrostatic pressures ( 10.0 mPa ) by the multifunctional thermostatic high-insensitive hydrostatic pressure loading device for 2 h per day lasting for 5 days. Toluidine blue staining and immunohistochemical staining of type II collagen were employed to identify the chondrocytes. Cell morphology was observed by light microscopy. The ultrastructure of chondrocytes were observed by transmission electron microscopy ( TEM ). Type II collagen immunohistochemical staining and semi-quantitative analysis were performed to measure contents and expression of type II collagen of chondrocytes in 2 groups. Results Compared with the control group, cell morphology changed from irregular polygon into long spindle, membrane and cytoplasm retracted, the number of cells was reduced significantly and grew sparsely in the 10.0 mPa group. TEM showed that various apoptosis indications such as uneven distribution and margination of chromosomes in the 10.0 mPa group. Collagen type II immunohistochemistry and semi-quantitative analysis showed that the percent of stained area of control group and 10.0 mPa group was ( 61.64 ± 6.19 ) % and ( 43.76 ± 5.61 ) %, compared with the control group, the stained area and extent of chondrocytes were significantly decreased in the 10.0 mPa group ( P < 0.05 ). Conclusions The high-intensity pressure over human physiological range can be regarded as a mechanical injury to result in cell apoptosis, morphology and intracellular ultrastructural changes of human chondrocytes, and also decrease protein expression of human chondrocytes. These data provide the experimental basis for studying the relationship between stress injury and pathogenesis of osteoarthritis.