HORMONAL CONTROL OF CHONDROCYTE CARTILAGE SYNTHESIS

这项研究的长期目标是描述 成体软骨基质的激素调节机制 关节软骨细胞的合成和组装。这个 实验目标是利用硫酸角蛋白的合成 蛋白多糖和II型胶原作为成人的标志物 体外无血清条件下软骨表型及测定 条件独特的结缔组织特异性组合 重建软骨基质所必需的激素。 01-03年度的研究结果显示，IGF- I/GH联合选择性刺激蛋白多糖合成 并提高培养物中硫酸角蛋白的水平。此外, 胰岛素、IGF-I和成纤维细胞生长因子的组合是 在确定的无血清介质中足以刺激 成年牛关节软骨细胞的增殖。 这项研究在04-06年度的延续将集中于 细胞外基质的合成和组装过程 关节软骨细胞在特定的培养环境中。这个 特定的目标都是基于荷尔蒙的假设。 控制软骨基质的合成，并针对 以下主题：(1)细胞外基质的分析 成人关节软骨细胞向融合生长的合成 在含有胰岛素、胰岛素样生长因子-I和成纤维细胞生长因子的无血清培养液中。(2) 研究结缔组织特异性激素的能力， 甲状旁腺激素，生长激素，甲状腺素，甲状腺降钙素， 雌激素和睾酮，以刺激基质合成 成熟表型作为胰岛素、胰岛素样生长因子-I和成纤维细胞生长因子的补充。(3) 调查激素、依恋因素、 蛋白水解酶抑制剂与细胞几何构型的关系 蛋白多糖转化为基质。(4)扩大血清条件- 人肋软骨细胞和关节软骨细胞的自由培养 量化培养基和细胞相关的特性 蛋白多糖。技术包括无血清软骨细胞 蛋白多糖的培养、定量和分离、分析 胶原蛋白的合成，糖胺聚糖的微量分析和 DNA和RNA的定量。 这些研究对骨科临床和临床具有重要意义。 重建外科；有关荷尔蒙的信息 控制软骨合成直接适用于抑制 软骨破坏，刺激软骨修复和 可移植软骨的制备。

The long-term objective of this research is to characterize the mechanisms of hormonal regulation of adult cartilage matrix synthesis and assembly by articular chondrocytes. The experimental goal is to use synthesis of keratan sulfate proteoglycans and Type II collagen as markers of the adult cartilage phenotype and to determine under in vitro serum-free conditions the unique combination of connective tissue specific hormones necessary for reconstruction of the cartilage matrix. Results obtained during years 01-03 of this grant show that IGF- I/GH in combination selectively stimulate proteoglycan sysnthesis and enhance levels of keratan sulfate in the culture. In addition, the combination of insulin, IGF-I and fibroblast growth factor are sufficient in a defined serum-free medium to stimulate the proliferation of adult bovine articular cartilage cells. The continuation of this research in years 04-06 will focus on the process of synthesis and assembly of the extracellular matrix by articular chondrocytes in a defined culture environment. The Specific Aims are all predicated on the hypothesis of hormonal control of cartilage matrix synthesis and are directed to the following topics: (1) Analysis of the extracellular matrix synthesized by adult articular cartilage cells grown to confluency in a serum-free medium containing insulin, IGF-I and FGF. (2) Investigate the ability of connective tissue specific hormones, parathyroid hormone, growth hormone, thyroxine, thyrocalcitonin, estrogen and testosterone, to stimulate matrix synthesis of a mature phenotype as adjuncts to insulin, IGF-I and FGF. (3) Investigate the influence of hormones, attachment factors, protease inhibitors and cellular geometry on the assembly of proteoglycans into matrix. (4) Extend the conditions for serum- free culture to human costal and articular chondrocytes and quantitate the characteristics of the medium and cell-associated proteoglycans. Techniques include serum-free chondrocyte culture, quantification and fractionation of proteoglycan, analysis of collagen synthesis, microanalysis of glycosaminoglycan and quantitation of DNA and RNA. The studies are significant to clinical orthopedics and reconstructive surgery; the information regarding hormonal control of cartilage synthesis is directly applicable to inhibition of cartilage destruction, stimulation of cartilage repair and preparation of transplantable cartilage.