人工关节炎性介质对滑膜细胞基质金属蛋白酶-9表达的影响

An experimental study on the effects of inflammatory media of artificial joint on synovial cell matrix metalloproteinase-9 expression

目的 观察人滑膜细胞基质金属蛋白酶-9（MMP-9）表达及活性受钛合金颗粒、白细胞介素-6（IL-6）、前列腺素E2（PGE2）的调控表现,探讨滑膜细胞在钛合金材料关节假体周围骨溶解的机制.方法 用钛合金颗粒、IL-6、PGE2加载滑膜细胞（关节滑膜细胞来源于正常人严重创伤后捐献,体外培养5代）,于12、24、48、72 h等时间节点收集上清液,检测MMP-9的活性（优化明胶酶谱法）.结果 直径最小的钛合金颗粒（≤5 μm）能刺激人滑膜细胞MMP-9蛋白表达是对照组的3.75倍;钛合金颗粒、IL-6、PGE2及其中两者联合作用致MMP-9高表达（P＜0.01）,三者联合作用有协同效应（P＜0.01）.钛合金颗粒、IL-6、PGE2能够刺激人关节滑膜细胞增加MMP-9蛋白活性,并具有协同叠加效应.结论 钛合金材料人工关节置换术后,磨损颗粒和IL-6、PGE2协同刺激滑膜细胞,胞外基质降解,导致假体的骨性静态支持结构力学性能下降,破骨细胞活性被激活,参与假体无菌性松动的发生.

Objective To research the human synovial cell matrix metalloproteinase 9 （MMP-9） expression and activity regulated by titanium alloy particles,interleukin （IL）-6 and prostaglandin E2 （PGE2） and discuss the mechanism of synovial cells for osteolysis around titanium alloy prosthesis.Methods The synovial cells of joint （from normal individuals＇ donation after serious trauma,in vitro culture of 5 generations） were loaded with titanium alloy particles,IL-6 and PGE2.The supernatants were collected at time points 12,24,48 and 72 h.The MMP-9 activity （optimized zymography） was detected.Results The stimulation of titanium alloy particles with minimum diameter （≤ 5 μm） on human synovial cell MMP-9 protein expression was 3.75 times that of the control group;the titanium alloy particles,IL-6,PGE2 and the combined effect of any two were associated with high expression of MMP-9 （P 〈 0.01） while a combination of all three were associated with synergistic effect （P 〈 0.01）.The titanium alloy particle,IL-6 and PGE2 could stimulate synovial cells of human joint to increase the MMP-9 protein activity,and had synergistic effect.Conclusion After total arthroplasty with titanium alloy artificial joint,the wear particles,IL-6 and PGE2 co-stimulate synovial cells,and the extracellular matrixes degrade to reduce the mechanical properties of static supporting bone structure of the prosthesis and participate in its aseptic loosening.