Characterization of cultured synovial cells from human TMJ

人颞下颌关节培养滑膜细胞的表征

基本信息

批准号：
11672017
负责人：
OGURA Naomi
金额：
$ 2.37万
依托单位：
Nihon University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1999
资助国家：
日本
起止时间：
1999 至 2001
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11672017/
关键词：
Cultured synovial cells TMJ IL-1β TNF-α Plasminogen activator IL-6 IL-8 MCP-1 顎関節症ヒト顎関節滑膜細胞

项目摘要

The aims of the present study were, firstly, to isolate and to characterize culture synovial cells obtained from human TMJ specimens, and secondly, to investigate the productions of cytokines and enzymes in those cells.<Methods> Synovial cells were isolated from TMJ synovial tissue using an out growth method and then cultured in Ham F12 containing 20% or 10% fetal bovine serum. Synovial cells were examined cell-specific markers using immunocytochemistry. In the next, synovial cells were treated with IL-1β or TNF-α, which was increased in synovial fluids from patients with TMJ disorders. Activity of plasminogen activator (PA), which is associated with extracellular matrix degradation, was measured by enzyme assay. Amounts of IL-6, IL-8 and macrophage attractant protein (MCP)-1 were measured by ELISA. Gene expressions of these cytokines were also investigated using RTPCR method.<Rersults> Synovial cells were positive for fibroblast-specific marker and vimentin. In contrast, the cells were negative for macrophage marker, HLA class II and dendritic cell marker. Then, synovial cells were treated with or without IL-1β. IL-1β increased IL-6, IL-8 and MCP-1 production in synovial cells in a time- and dose-dependent manner. IL-1β stimulated gene expression of these cytokines in them. IL-1β also enhanced PA activity in synovial cells. In the next, the cells were treated with TNF-α. TNF-α enhanced productions and gene expressions of IL-8 and MCP-1.<Conclusions> We found that cultured synovial cells might provide important advantages for studies of the cellular and molecular mechanisms in TMJ. In addition, PA activity and IL-6, IL-8, and MCP-1 productions may be increased in synovial cells when IL-1β and TNF-α levels are increased in synovial fluids from patients with TMJ disorders.

首先，本研究的目的是分离并表征从人TMJ样品中获得的培养滑膜细胞，其次，研究这些细胞中细胞因子和酶的生产。<方法>滑膜细胞是从TMJ Synovial组织中使用OUT生长方法从TMJ Synovial组织中分离出来的，然后使用HAM F12含有20％或10％的FTAIM covine培养。使用免疫细胞化学检查滑细胞特异性标记。接下来，将滑膜细胞用IL-1β或TNF-α处理，TMJ疾病患者的滑液中增加了。通过酶测定测量纤溶酶原激活剂（PA）的活性，与细胞外基质降解相关。通过ELISA测量IL-6，IL-8和巨噬细胞吸引蛋白（MCP）-1的量。还使用RTPCR方法研究了这些细胞因子的基因表达。<rersults>滑膜细胞对成纤维细胞特异性标记和波形蛋白呈阳性。相反，细胞对巨噬细胞标记，HLA II类和树突状细胞标记为阴性。然后，将滑膜细胞用或没有IL-1β处理。 IL-1β在滑膜细胞中以时间和剂量依赖性方式增加了IL-6，IL-8和MCP-1的产生。 IL-1β刺激了这些细胞因子中这些细胞因子的基因表达。 IL-1β还增强了滑膜细胞中的PA活性。接下来，将细胞用TNF-α处理。 TNF-α增强了IL-8和MCP-1的生产和基因表达。<结论>我们发现，培养的滑膜细胞可能为研究TMJ中细胞和分子机制的研究提供重要优势。此外，当来自TMJ疾病患者的滑液中IL-1β和TNF-α水平升高时，滑膜细胞中PA活性和IL-6，IL-8和MCP-1生产可能会增加。