ACTIVATION OF HUMAN CHONDROCYTES

本提案中描述的研究旨在调查控制措施 阿司匹林的产物激活软骨细胞的机制 炎症及其在细胞降解和修复中的作用 软骨基质。建立了细胞培养模型。 单层培养的人关节软骨细胞作为靶细胞 培养的人血单核细胞的可溶性产物的作用， 单核细胞因子(MCF)，刺激产生 前列腺素和中性蛋白酶。我计划进一步描述这一点 细胞相互作用，使用胶原合成作为基质的衡量标准 修复和纤溶酶原激活剂作为降解的衡量标准。人类 琼脂基质中的软骨细胞培养将被建立，并将 已确定这些细胞是否能够被MCF在以下条件下激活 这些条件。琼脂糖合成的胶原蛋白类型比较 将对单层培养进行表征，并研究MCF对 我们将研究II型与I型胶原的合成。我打算 软骨细胞纤溶酶原激活物调控的继续研究 比较细胞与释放的活动相关的培养和 进一步鉴定了丝氨酸蛋白酶的抑制物--蛋白连接蛋白， 其释放由软骨细胞由MCF刺激。无单元翻译 将利用从MCF刺激的软骨细胞中提取的mRNA来研究 特定软骨细胞蛋白的可翻译RNA水平。该控件 在转录水平上产生I型和II型胶原蛋白 水平将使用特定的人胶原基因探针进行研究。最后， 这些研究最终将导致对许可作用的调查 生长和附着因子在软骨细胞合成功能中的作用。

Studies described in this proposal are designed to investigate the control mechanisms involved in the activation of chondrocytes by products of inflammation and the role of this process in the degradation and repair of cartilage matrix. A cell culture model has been established employing human articular chondrocytes in monolayer culture as target cells for the action of a soluble product from cultured human blood mononuclear cells, mononuclear cell factor (MCF), which stimulates the production of prostaglandins and neutral proteases. I plan to characterize further this cellular interaction, using collagen synthesis as a measure of matrix repair and plasminogen activator as a measure of degradation. Human chondorcyte cultures in agarose matrix will be established and it will be determined whether these cells are capable of being activated by MCF under these conditions. The types of collagen synthesized in agarose compared with monolayer cultures will be characterized and the effects of MCF on type II vs. type I collagen synthesis will be studied. I intend to continue studies of the control of plasminogen activator in chondrocyte cultures comparing cell-associated with released activity and to characterize further an inhibitor of serine proteasses, protease nexin, whose release by chondrocytes is stimulated by MCF. Cell-free translation of mRNA from MCF-stimulated chondrocytes will be employed to study the levels of translatable RNA for specific chondorcyte proteins. The control of the production of type I and type II collagens at the transcriptional level will be studied using specific human collagen cDNA probes. Finally, these studies will lead eventually to investigation of the permissive role of growth and attachment factors in chondrocyte synthetic functions.