BONE-MATRIX RGD GLYCOPROTEINS - IMMUNOLOCALIZATION AND INTERACTION WITH HUMAN PRIMARY OSTEOBLASTIC BONE-CELLS IN-VITRO
BONE-MATRIX RGD GLYCOPROTEINS - IMMUNOLOCALIZATION AND INTERACTION WITH HUMAN PRIMARY OSTEOBLASTIC BONE-CELLS IN-VITRO
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DOI：
10.1002/jbmr.5650090408
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发表时间：
1994-04-01
影响因子：
6.2
通讯作者：
ROBEY, PG
中科院分区：
文献类型：
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作者：
GRZESIK, WJ;ROBEY, PG
细胞与细胞外基质的相互作用对于它们的锚定、增殖、迁移和分化至关重要。骨基质中存在多种含有整合素结合 RGD 序列的糖蛋白：纤连蛋白 (FN)、血小板反应蛋白 (TSP)、骨桥蛋白 (OPN)、骨唾液蛋白 (BSP)、I 型胶原 (COLL 1) 和玻连蛋白 (VN)。在本研究中，使用免疫组织化学方法检查了 TSP、FN、VN 和几种整合素在发育中的人长骨中的定位，以及所有骨 RGD 蛋白对人成骨细胞粘附的影响。血小板反应蛋白、纤连蛋白和玻连蛋白在骨组织内显示出不同的定位模式。 TSP主要存在于类骨质和骨膜中； VN 似乎主要存在于成熟骨基质中。 FN 存在于骨膜以及成熟和未成熟骨基质中。使用一组抗整合素抗体，我们发现骨细胞在体内和体外表达α4、α(v)、α5β1、α(v)β3和β3/β5整合素，并且这些受体大部分在不同成熟阶段的所有骨细胞上表达，具有定量而不是定性变化，但α4除外，α4是 主要由成骨细胞表达。在无血清条件下使用成骨细胞谱系的原代人细胞进行细胞附着测定。 COLL 1、TSP、VN、FN、OPN 和 BSP 以剂量依赖性方式促进骨细胞附着，并且在等摩尔浓度下使用时作用相同。当培养基中存在 GRGDS 肽时，对 BSP、OPN 和 VN 的粘附几乎完全被阻断（分别为对照的 10%、10% 和 15%），以及对 FN、COLI 的附着。 1，TSP 仅略有下降（分别为 80%、75% 和 55%）。这些结果表明，人类骨细胞可能使用不依赖于 RGD 的机制来附着到后者的糖蛋白上。
The interaction of cells with extracellular matrix is essential for their anchorage, proliferation, migration, and differentiation. In bone matrix there are multiple glycoproteins that contain the integrin-binding RGD sequence: fibronectin (FN), thrombospondin (TSP), osteopontin (OPN), bone sialoprotein (BSP), type I collagen (COLL 1), and vitronectin (VN). In this study, the localization of TSP, FN, VN, and several integrins within developing human long bone using immunohistochemical methods was examined, as was the effect of all bone RGD proteins on the adhesion of human osteoblastic cells. Thrombospondin, fibronectin, and vitronectin showed distinct localization patterns within bone tissue. TSP was found mainly in osteoid and the periosteum; VN appeared to be present mainly in mature bone matrix. FN was present in the periosteum as well as within both mature and immature bone matrix. Using a panel of antiintegrin antibodies we found that bone cells in vivo and in vitro express alpha4, alpha(v), alpha5beta1, alpha(v)beta3, and beta3/beta5 integrins, and these receptors are for the most part expressed on all bone cells at different stages of maturation with quantitative rather than qualitative variations, with the exception of alpha4, which is expressed mainly by osteoblasts. Cell attachment assays were performed using primary human cells of the osteoblastic lineage under serum-free conditions. COLL 1, TSP, VN, FN, OPN, and BSP promoted bone cell attachment in a dose-dependent manner and were equivalent in action when used in equimolar concentrations. In the presence of GRGDS peptide in the medium, the adhesion to BSP, OPN, and VN was almost completely blocked (10, 10, and 15% of control, respectively), and attachment to FN, COLI. 1, and TSP was only slightly decreased (80, 75, and 55 %, respectively). These results suggest that human bone cells may use RGD-independent mechanisms for attachment to the latter glycoproteins.