钛颗粒和TNF-α对人膝关节滑膜细胞MMP-1、2、3表达的影响  

Effect of Titanium Particles and TNF-αon the Gene Expression and Activity of MMP-1,2,3 in Human Knee Joint Synovial Cells

作　　者：符纯锋[1] 谢静[2] 陈荣富[1] 王春莉[2] 许春明[2] 陈诚[1] 王志强[1] 林良波[1] 黄伟[1] 梁熙[1] 宋国立[2,3] 

机构地区：[1]重庆医科大学附属第一医院骨科,重庆400016 [2]重庆大学生物工程学院,重庆400044 [3]加州大学圣迭亚哥分校骨科系及生物工程系

出　　处：《生物医学工程学杂志》2013年第5期1022-1026,共5页Journal of Biomedical Engineering

基　　金：重庆市卫生局医学科研资助项目(2011-2-054)

摘　　要：研究钛颗粒和肿瘤坏死因子(TNF-α)对人滑膜细胞基质金属蛋白酶(MMP-1、2、3)表达及活性的影响,从而探讨滑膜在钛磨损颗粒诱导的金对金人工关节假体周围骨溶解中的可能机制。用钛颗粒和TNF-α处理体外培养的人膝关节滑膜细胞,在处理后2h提取总RNA;逆转录PCR、荧光实时定量PCR对人滑膜细胞MMP-1、2、3基因的表达进行半定量和定量分析。处理后12、24和48h收集培养上清液,用明胶酶谱法检测MMP-2的活性。结果显示,与对照组相比,钛颗粒、TNF-α均明显增加了MMP-1、2、3基因的表达(P<0.05),两者联合作用更明显(P<0.01)。MMP-2蛋白水平和基因表达趋势基本一致,钛颗粒和TNF-α处理分别增加MMP-2活性1.3倍和1.5倍(P<0.05),联合处理作用更为明显,增加1.7倍(P<0.01)。钛颗粒和TNF-α能够刺激人膝关节滑膜细胞MMP-1、2、3基因表达的增加以及MMP-2蛋白活性。本研究表明,金对金人工关节置换术后,磨损钛颗粒可能刺激滑膜细胞,增加骨细胞外基质的降解,导致假体的骨性支持结构力学性能下降,促使假体无菌性松动的发生。This paper is aimed to investigate the effect of titanium （Ti）particles and tumor necrosis factor α（TNFα） on the expressions of MMP-1, 2, 3 in human synovial cells, so as to explore the possible mechanism of osteolysis post-operation of metal on-metal total joint arthroplasty ;human synovial cells induced by Ti particles. In vitro cell cultures, human synovial cells were treated by Ti particles and/or TNF-α The total RNA was isolated at 2 hours af- ter the treatment. The gene expression of MMP-1, 2, 3 was analyzed by Semi-quantitative Reverse-transcriptiona; PCR and quantitative real time PCR. Cell supernatant was collected at 12, 24, 48 hours after the treatment and Ge latin zymography was performed to detect the activity of MMP-2. Compared to those in the control group （untreat ed）, Ti particles and TNF-a increased the gene expression of MMP 1, 2, 3 respectively （P〈0.05）, and the effect of combination of the two was even more significant （P〈0.01）. The trend of activities of MMP-2 is similar with gene expression. Ti particles and TNF α increased MMP-2 activities by 1.3 times and 1.5 times respectively （P〈0.05）, and the combination of the two increased by 1.7 times （P〈0.01）. Ti particles and TNF-α-induced the stimulation of MMP-1, 2, 3 expressions and MMP-2 activities in human knee joint synovial cells may be involved in aseptic loose- ning after metal-on-metal arthroplasty through increasing the degradation of bone matrix and declining of osseous support structure mechanics.

关 键 词：钛颗粒 无菌性松动 滑膜 肿瘤坏死因子 基质金属蛋白酶 

分 类 号：R684[医药卫生—骨科学]