Response of human osteoblasts to implant materials: Integrin-mediated adhesion
Response of human osteoblasts to implant materials: Integrin-mediated adhesion
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DOI：
10.1002/jor.1100140606
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发表时间：
1996-11-01
影响因子：
2.8
通讯作者：
McCarthy, MB
中科院分区：
文献类型：
--
作者：
Gronowicz, G;McCarthy, MB
分析了人成骨细胞样细胞系 Saos-2 与骨科植入材料的初始相互作用，以确定这些细胞粘附到植入物表面的机制。 Saos-2 细胞被允许附着在由骨科植入材料 Tivanium (Ti6Al4V) 和 Zimaloy (CoCrMo) 组成的圆盘上，并控制玻璃和塑料圆盘。血清对 4 小时或 24 小时时附着在 Tivanium 和 Zimaloy 上的细胞数量没有影响，但确实增加了 24 小时时附着在玻璃上的细胞数量。通过[H-3]脯氨酸掺入胶原酶可消化的蛋白质和非胶原蛋白来测定胶原合成。与玻璃相比，在 Zimaloy 上培养 24 小时的细胞中合成的胶原蛋白显着增加了 19%，而 Tivanium 和塑料没有差异。然而，与玻璃相比，Tivanium 上的胶原酶可消化蛋白质和非胶原蛋白增加最多（分别为 204% 和 198%）。为了确定整合素是否参与细胞与植入材料的附着，将肽 GRGDSP (Gly-Arg-Gly-Asp-Ser-Pro) 添加到无血清培养基中的细胞中，该肽通过 Arg-Gly-Asp 序列阻断整合素受体。该肽在 Tivanium 上抑制细胞粘附 28%，在 Zimaloy 上抑制细胞粘附 40%，但对玻璃和塑料没有影响。对照肽 GRADSP (Gly-Arg-Ala-Asp-Ser-Pro) 对粘附没有影响。蛋白质合成的抑制和表面蛋白质的酶促去除并不影响Arg-Gly-Asp肽抑制细胞附着至植入材料的能力。这些结果表明整合素能够直接与 Tivanium 和 Zimaloy 结合。整合素蛋白的蛋白质印迹分析表明，许多整合素亚基发生变化，具体取决于细胞附着的底物。特别是，β(1) 整合素亚基在 24 小时内增加了 3.8 至 9.5 倍。为了具体确定哪些整合素可能参与粘附，添加了整合素抗体。纤连蛋白受体 α(5) beta(1) 的抗体可显着抑制细胞与 Tivanium 的结合 63%，以及与 Zimaloy 的结合 49%，并且对玻璃没有影响。玻连蛋白受体抗体 alpha(v) beta(3)/beta(5) 不会改变细胞粘附。总之，成骨细胞样细胞似乎能够通过整合素直接附着到植入材料上。底物的类型决定了成骨细胞表达哪些整合素和细胞外基质蛋白。这些数据提供了有关植入材料如何影响成骨细胞分化和骨生长的信息。
The initial interaction of the human osteoblast-like cell line Saos-2 with orthopaedic implant materials was analyzed to determine the mechanism by which these cells adhere to implant surfaces. Saos-2 cells were allowed to attach to disks composed of the orthopaedic implant materials Tivanium (Ti6Al4V) and Zimaloy (CoCrMo) and to control disks of glass and plastic. Serum had no effect on the number of cells that attached to Tivanium and Zimaloy at 4 or 24 hours but did increase the number of cells that attached to glass at 24 hours. Collagen synthesis was determined by [H-3]proline incorporation into collagenase-digestible protein and noncollagen protein. A significant increase of 19% was found for collagen synthesized in cells cultured on Zimaloy for 24 hours compared with glass, with no differences on Tivanium and plastic. However, collagenase-digestible protein and noncollagen protein were increased the most (204 and 198%, respectively) on Tivanium compared with glass. To determine if integrins were involved in cell attachment to implant materials, the peptide GRGDSP (Gly-Arg-Gly-Asp-Ser-Pro), which blocks integrin receptors through the Arg-Gly-Asp sequence, was added to the cells in serum-free medium. This peptide inhibited cell adhesion by 28% on Tivanium and 40% on Zimaloy but had no effect on glass and plastic. The control peptide GRADSP (Gly-Arg-Ala-Asp-Ser-Pro) had no effect on adhesion. Inhibition of protein synthesis and enzymatic removal of surface proteins did not affect the ability of Arg-Gly-Asp peptides to inhibit cell attachment to the implant materials. These results suggest that integrins are able to bind directly to Tivanium and Zimaloy. Western blot analysis of integrin protein demonstrated changes in many integrin subunits, depending on the substrate to which cells attached. In particular, the beta(1) integrin subunit was increased 3.8 to 9.5-fold at 24 hours. To determine specifically which integrins may be involved in adhesion, antibodies to integrins were added. An antibody to the fibronectin receptor, alpha(5) beta(1), significantly inhibited binding of cells to Tivanium by 63% and to Zimaloy by 49% and had no effect on glass. The vitronectin receptor antibody, alpha(v) beta(3)/beta(5), did not alter cell adhesion. In conclusion, osteoblast-like cells appear to be capable of attaching directly to implant materials through integrins. The type of substrate determines which integrins and extracellular matrix proteins are expressed by osteoblasts. These data provide information on how implant materials may affect osteoblast differentiation and bone growth.