MINERALIZATION STUDIES RELATED TO ORAL BIOLOGY

与口腔生物学相关的矿化研究

• 批准号: 2129070
• 负责人: HARRISON CLARKE ANDERSON
• 金额: $ 23.46万
• 依托单位: UNIVERSITY OF KANSAS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 1978
• 资助国家: 美国
• 起止时间: 1978-03-01 至 1998-02-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2129070
• 关键词: 1,25 dihydroxycholecalciferol; SDS polyacrylamide gel electrophoresis; alkaline phosphatase; ascorbate; calcification; calcitonin; cartilage; cell growth regulation; chondrocytes; dental development; dentin; dexamethasone; electron microscopy; enzyme activity; extracellular matrix proteins; fluorine; glucocorticoids; immunocytochemistry; laboratory rat; membrane biogenesis; membrane proteins; osteoblasts; retinoate; thyroxine; tissue /cell culture; transforming growth factors; vesicle /vacuole; western blottings

The proposed study will focus on matrix vesicles (MVs) which play an initiating role in the mineralization of teeth and bones. MVs are submicroscopic, extracellular, membrane-invested particles that serve as the initial site of calcification in dentin, growth cartilage and developing bone. Our lab was involved in the first identification, isolation and characterization of MVs. We and others have provided evidence that MV phosphatases, including alkaline phosphatase (ALP), are involved in MV mineralization. Furthermore, these phosphatases are integrated into the MV membrane under which mineral first appears suggesting a critical role for ALP and other components of the MV membrane in initiating calcification. Our specific aims are directed toward an increased understanding of the molecular and structural organization of the MV membrane and sap with emphasis on identifying and localizing major constitutive proteins, testing the function of MV phosphatases in the calcification mechanism of isolated MVs, and studying the regulation of MV biogenesis by cultured bone cells and marrow osteoprogenitor cells. Proposed new studies include: 1) further analysis of the composition of isolated MVs to complete the identification of major proteins; 2) assignment of major MV constituents to the membrane versus the vesicle sap using high resolution EM immunolocalization coupled with biochemical release of membrane-attached proteins by surface active agents versus total release by detergents; 3) further characterization of the role of MV phosphatases in the mineralization of MVs from whole cartilage versus MVs generated in cultures of bone cells or marrow osteoprogenitor cells. Agents such as 1,250H2D3, dexamethasone, or fluoride, known to boost MV ALP activity, will be tested for a positive effect on MV calcification; 4) further studies will characterize the mechanism of MV biogenesis by cultured rat chondrocytes, osteoblasts and osteoprogenitor cells of the marrow stroma as modified by agents known to promote osseous differentiation in vitro including glucocorticoids, fluoride and retinoic acid. This is a fundamental study of the mechanism by which dental and skeletal forms of mineralization are initiated. New knowledge of the matrix vesicle calcification can be applied to a broad range of topics including specific diseases in which abnormal calcification occurs.

拟议的研究将重点介绍发挥作用的基质囊泡（MV） 在牙齿和骨骼的矿化中发挥作用。 MV是 亚镜，细胞外，膜侵入的颗粒，用作 牙本质中钙化的初始位点，生长软骨和 发育骨骼。 我们的实验室参与了第一个识别， MV的隔离和表征。 我们和其他人提供了 证据表明包括碱性磷酸酶（ALP）在内的MV磷酸酶是 参与MV矿化。 此外，这些磷酸酶是 集成到MV膜中，首先出现矿物质 提出了MV的ALP和其他组件的关键作用 启动钙化时的膜。 我们的具体目标是指导的 促进对分子和结构的了解 MV膜和SAP的组织，重点是识别和 定位主要的组成蛋白，测试MV的功能 分离的MV的钙化机理中的磷酸酶，并研究 通过培养的骨细胞和骨髓调节MV生物发生 骨基因细胞。 拟议的新研究包括：1）进一步分析 孤立的MV以完成主要蛋白质的鉴定； 2） 将主要MV成分分配给膜与囊泡 使用高分辨率EM免疫定位与生化的SAP 通过表面活性剂而不是释放膜连接蛋白 洗涤剂的总释放； 3）进一步表征 MV的MV磷酸酶从整个软骨的MV矿化中 MVS在骨细胞或骨髓骨基元细胞的培养物中产生。 诸如1,250H2D3，地塞米松或氟化物之类的代理，已知可以增强MV ALP活性将测试对MV钙化产生积极影响； 4）进一步的研究将表征MV生物发生的机理 培养的大鼠软骨细胞，成骨细胞和骨源细胞的细胞 骨髓基质是由已知促进骨的试剂修饰的 体外分化包括糖皮质激素，氟化物和视黄 酸。 这是对牙齿和骨骼的机制的基本研究 开始矿化形式。 矩阵的新知识 囊泡钙化可以应用于广泛的主题 发生异常钙化的特定疾病。