REGULATION OF COLLAGEN MATRIX MINERALIZATION

描述(申请人提供)：本研究的长期目标是了解骨和牙齿中胶原基质矿化的调节机制。在上一次资助期间，我们已经部分表征了胶原交联的独特方面，以及矿化组织中胶原结合富含亮氨酸的小分子蛋白多糖(CB-SLRPs)的特定组织分布和mRNA表达模式。这些和其他发现引导我们假设：1.I型胶原的共价分子间交联参与调节矿化的空间方面；2.胶原结合蛋白多糖(DCN)部分调节胶原矿化的时间和质量。为了检验这些假设，我们想要：1。建立合成低水平赖氨酰羟基酶-2的成骨细胞克隆，以将交联途径从矿化组织表型切换到非矿化组织表型，1b。鉴定胶原基质的赖氨酸羟化和交联化学(类型、数量和分子分布)，以及这些克隆在体内外产生的矿化模式。2A。通过在成骨细胞系中的过表达和反义方法，研究DCN在体内外胶原成熟和矿化中的作用。2B。研究DCN缺陷和DCN/Biglycan-Double缺陷小鼠骨/牙本质中基质矿化(胶原和矿物质)的特征。2C。探讨DCN与胶原相互作用对胶原体外成熟的影响。这项研究获得的数据可能为深入了解骨骼和牙齿中胶原矿化的调控机制提供依据。

DESCRIPTION (provided by applicant): The long-term goal of this study is to understand the regulatory mechanisms of collagen matrix mineralization in bones and teeth. During the last grant period, we have partially characterized the unique aspects of collagen cross-linking, and specific tissue distribution as well as mRNA expression pattern of collagen-binding small leucine-rich proteoglycans (CB-SLRPs) in mineralized tissues. These and other findings have led us to hypothesize: 1. the covalent intermolecular cross-linking of type I collagen is involved in regulating the spatial aspect of mineralization, and 2. a collagen-binding proteoglycan, decorin (DCN), partially regulates the timing and quality of collagen mineralization. To test these hypotheses, we would like: 1a. To establish osteoblastic cell clones that synthesize low levels of lysyl hydroxylase-2 to switch the cross-linking pathway from mineralized to non-mineralized tissue phenotype, 1b. To characterize the lysine hydroxylation and cross-linking chemistries (type, quantity and molecular distribution) of collagen matrix, and the mineralization pattern produced by the clones in vitro and in vivo. 2a. To investigate the roles of DCN in collagen maturation and mineralization in vitro and in vivo by employing overexpression and antisense approaches in osteoblastic cell line. 2b. To characterize the matrix mineralization (collagen and mineral) in bones/dentin in DCN-deficient and DCN/biglycan-double deficient mice. 2c. To evaluate the effects of DCN-collagen interaction on collagen maturation in vitro. The data obtained from this study may provide insights into the regulatory mechanisms of collagen mineralization in bones and teeth.