循环牵张应力对人骨关节炎软骨细胞增殖的影响

【摘要】目的探讨循环牵张应力对体外培养的人骨关节炎患者软骨细胞增殖的影响。方法退变软骨来源于1例66岁骨关节炎患者的手术取材，病理学诊断评估其退变程度。无菌条件下酶消化法原代培养退变软骨细胞，并用甲苯胺蓝、番红O和Ⅱ型胶原免疫荧光染色进行鉴定。将第3代细胞接种于硅橡胶膜载体上，用Electro?Force3200力学试验仪搭载的BioDynamic生物反应仓对其施加0、5%、10%和15%的应变作用3h，受力完成后继续培养24h，用流式细胞仪测定不同力学环境下细胞的增殖活性。结果随着牵张应力的增加，S期细胞百分比和增殖指数（PI）呈缓慢上升趋势，10%时达到峰值，之后开始下降。结论体外培养的人骨关节炎软骨细胞的增殖活性与受力大小有关，且存在细胞增殖的最适应力条件；超过此应力范围其增殖活性将会降低。

Effects of Cyclic Tensile Strain on Proliferation of Human Osteoarthritis Chondrocytes

[Abstract]ObjectiveTo explore the effects of cyclic tensile strain on the proliferation of cultured human osteoar -thritis chondrocytes.MethodsThe degenerated human cartilage came from a66-year-old patient with osteoarthritis. The degree of degeneration was assessed by pathological diagnosis. Under the sterile condition, degenerated chondrocytes werecultured by enzymatic digestion. The toluidine blue staining, safranin O staining, and immunofluorescence staining of colla-gen Ⅱ were used to identify the cultured cells. The third generation cells were seeded on the silicone rubber membrane carrier, using the EF3200mechanical tester equipped with BioDynamic bioreactor system, imposed a frequency of 0.25Hz, stain 0, 5%,10% and 15% load for 3h. The proliferation activity was detected by flow cytometry.ResultsThe proliferative in-dex (PI) increased with the magnitude value of cyclic tensile strain. The most significant increase of proliferation index was found in10% group.Conclusion The proliferation of human osteoarthritis chondrocytes increases under some stress