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首页> 外文学位 >The quantitative assessment of osteoinductivity of human demineralized bone matrix and cDNA array analysis of osteogenic differentiation in human periosteal cells.
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The quantitative assessment of osteoinductivity of human demineralized bone matrix and cDNA array analysis of osteogenic differentiation in human periosteal cells.

机译：人脱矿质骨基质骨诱导性的定量评估和人骨膜细胞成骨分化的cDNA阵列分析。

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摘要

Demineralized bone matrix (DBM) has been extensively utilized in orthopeadic, periodontal, and maxillofacial applications. DBM comprises bone morphogenetic proteins (BMPs) that are essential regulators for endochondral bone formation. The current study was undertaken to establish the quantitative in vitro assays for determining the osteoinductive potential of human DBM and the use of cDNA gene expression array technology for examining gene expression profiles during osteoblast differentiation in human periosteal cells.; BMP-4 is one of the most osteoinductive factors and has been investigated for clinical applications. For this reason, BMP-4 was designated as the osteoinductive protein for use in the quantitative in vitro assay. At the same time, the osteoinductivity of DBM was measured by histomorphometric analysis of new bone formation in an in vivo athymic mouse bioassay. DBM exhibiting high osteoinductivity in the nude mouse bioassay contained higher amounts of extractable BMP-4 than did DBM samples possessing low osteoinductivity.; In the study of the effect of residual calcium on extractability of BMP-4, the extractable BMP-4 content was undetectable in nondemineralized bone matrix. As the residual calcium level decreased, the extractability of BMP-4 in DBM increased. DBM particles less than 250 microns yielded the lowest amounts of extractable BMP-4 among all size groups tested. Bone particles in the 500–710 micron size range provided for DBM with the highest extractability of BMP-4. In the donor age study, the BMP-4 content in the protein extracts of DBM appears to be age-dependent, with DBM from younger donors being most likely to have greater BMP-4 quantity.; In the in vitro dose-response studies, DBM-conditioned media (DBM-CM) enhanced alkaline phosphatase activities of human periosteal cells in a dose-dependent fashion. Alkaline phosphatase activities in both biochemical and histochemical analysis increased in the DBM-CM treated flasks compared to non-treated flasks after five days of incubation. In gene expression analysis of osteogenic differentiation using cDNA array analysis, a few differentially expressed genes, including biglycan, TGF-β1, and TGF-βR1 were up-regulated, whereas collagen14A1 was down-regulated in response to DBM-CM treatment. In the present study, gene expression array technology provides insight to the biological process of osteogenic differentiation.
机译：脱矿质骨基质（DBM）已在矫形，牙周和颌面应用中广泛使用。 DBM包含骨形态发生蛋白（BMP），它们是软骨内骨形成的重要调节剂。目前的研究是建立用于确定人DBM骨诱导潜力的定量体外分析方法，以及使用cDNA基因表达阵列技术检查人骨膜细胞在成骨细胞分化过程中的基因表达谱。 BMP-4是最能诱导骨形成的因子之一，已被研究用于临床。因此，BMP-4被指定为用于体外定量分析的骨诱导蛋白。同时，在体内无胸腺小鼠生物测定法中，通过组织形态计量学分析新骨形成来测量DBM的骨诱导性。在裸鼠生物测定中显示出高骨诱导性的DBM比具有低骨诱导性的DBM样品含有更多的可提取BMP-4。在研究残余钙对BMP-4可提取性的影响时，在未脱矿化的骨基质中未检测到可提取BMP-4的含量。随着残留钙水平的降低，BMP-4在DBM中的可萃取性增加。在所有测试的尺寸组中，小于250微米的DBM颗粒产生的可提取BMP-4量最低。 500-710微米尺寸范围内的骨颗粒为DBM提供了BMP-4最高的可萃取性。在供体年龄研究中，DBM蛋白质提取物中的BMP-4含量似乎与年龄有关，年轻供体的DBM最有可能具有更大的BMP-4含量。在体外剂量反应研究中，DBM条件培养基（DBM-CM）以剂量依赖性方式增强了人骨膜细胞的碱性磷酸酶活性。孵育五天后，与未处理的烧瓶相比，DBM-CM处理的烧瓶在生化和组织化学分析中的碱性磷酸酶活性均增加。在使用cDNA阵列分析进行成骨分化的基因表达分析中，响应于DBM-CM处理，一些差异表达的基因（包括双糖链蛋白聚糖，TGF-β1和TGF-βR1）被上调，而胶原蛋白14A1被下调。在本研究中，基因表达阵列技术为成骨分化的生物学过程提供了见识。

著录项

  • 作者

    Honsawek, Sittisak.;

  • 作者单位

    Old Dominion University.;

  • 授予单位 Old Dominion University.;
  • 学科 Biology Molecular.; Biology Cell.
  • 学位 Ph.D.
  • 年度 2003
  • 页码 123 p.
  • 总页数 123
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 分子遗传学;细胞生物学;
  • 关键词

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