Intermittent hydrostatic pressure inhibits matrix metalloproteinase and pro-inflammatory mediator release from human osteoarthritic chondrocytes in vitro.
Intermittent hydrostatic pressure inhibits matrix metalloproteinase and pro-inflammatory mediator release from human osteoarthritic chondrocytes in vitro.
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DOI：
10.1016/j.joca.2004.05.008
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发表时间：
2004-09
影响因子：
7
通讯作者：
M. Trindade;J. Shida;T. Ikenoue;Mel S Lee;Eric Y. Lin;B. Yaszay;S. Yerby;S. Goodman;D. Schurman;R. L. Smith
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文献类型：
--
作者：
M. Trindade;J. Shida;T. Ikenoue;Mel S Lee;Eric Y. Lin;B. Yaszay;S. Yerby;S. Goodman;D. Schurman;R. L. Smith
目的验证骨关节炎软骨细胞间歇静水压力对基质金属蛋白酶和促炎介质体外释放的调节作用。设计分离培养人骨关节炎关节软骨细胞为原代高密度单层细胞。为了进行测试，将软骨细胞培养物转移到无血清培养基中，并在不加载或暴露于10MPa、频率为1Hz的间歇静水压力（IHP）下维持6、12和24小时。ELISA法检测基质金属蛋白酶-2、-9 （MMP-2、-9）、组织金属蛋白酶-1抑制剂（TIMP-1）、促炎介质白介素-6 （IL-6）、单核细胞趋化蛋白-1 （MCP-1）释放水平。酶谱分析证实基质金属蛋白酶活性。结果在没有IHP的情况下，软骨细胞培养液中MMP-2、TIMP-1、IL-6和MCP-1水平呈时间依赖性升高。在测试的所有时间段，相对于保持相同时间段的未加载对照培养物，IHP的应用降低了MMP-2水平。虽然84/82kDa带可以通过酶谱检测到，但通过酶联免疫吸附试验（ELISA）无法定量培养基中MMP-9的水平。TIMP-1水平在任何测试期间均未因IHP而改变。与保持相同时间的对照培养物相比，暴露于IHP的培养物在12和24小时时IL-6和MCP-1水平下降。结论体外实验中，ihp降低骨关节炎软骨细胞MMP-2、IL-6和MCP-1的释放，提示压力通过调节软骨细胞中这些降解和促炎蛋白的表达来影响软骨的稳定性。
OBJECTIVEThis study tested the hypothesis that intermittent hydrostatic pressure applied to human osteoarthritic chondrocytes modulates matrix metalloproteinase and pro-inflammatory mediator release in vitro.DESIGNHuman osteoarthritic articular chondrocytes were isolated and cultured as primary high-density monolayers. For testing, chondrocyte cultures were transferred to serum-free medium and maintained without loading or with exposure to intermittent hydrostatic pressure (IHP) at 10MPa at a frequency of 1Hz for periods of 6, 12 and 24h. Levels of matrix metalloproteinase-2, -9 (MMP-2, -9), tissue inhibitor of metalloproteinase-1 (TIMP-1), and the pro-inflammatory mediators, interleukin-6 (IL-6) and monocyte chemoattractant protein-1 (MCP-1), released into the culture medium were assessed by ELISA. Matrix metalloproteinase activity was confirmed by zymographic analysis.RESULTSIn the absence of IHP, levels of MMP-2, TIMP-1, IL-6, and MCP-1 in the chondrocyte culture medium increased in a time-dependent manner. Application of IHP decreased MMP-2 levels at all time periods tested, relative to unloaded control cultures maintained for the same time periods. Although 84/82kDa bands were faintly detectable by zymography, MMP-9 levels were not quantifiable in medium from loaded or unloaded cultures by ELISA. TIMP-1 levels were not altered in response to IHP at any time period tested. IL-6 and MCP-1 levels decreased in cultures exposed to IHP at 12 and 24h, relative to unloaded control cultures maintained for the same time periods.CONCLUSIONIHP decreased release of MMP-2, IL-6 and MCP-1 by osteoarthritic chondrocytes in vitro suggesting that pressure influences cartilage stability by modulating chondrocyte expression of these degradative and pro-inflammatory proteins in vivo.