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首页> 外文期刊>Acta biomaterialia >Fabrication and characterization of a recombinant fibronectin/cadherin bio-inspired ceramic surface and its influence on adhesion and ossification in vitro.
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Fabrication and characterization of a recombinant fibronectin/cadherin bio-inspired ceramic surface and its influence on adhesion and ossification in vitro.

机译：重组纤连蛋白/钙粘蛋白生物启发陶瓷表面的制备，表征及其对体外黏附和骨化的影响。

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摘要

This study has investigated the effects of a bio-inspired ceramic surface modified with a novel recombinant protein on surface parameters and cell behavior. The surface of a biphasic calcium phosphate ceramic was functionalized with a recombinant protein spanning the fragments of fibronectin module III7-10 and extracellular domains 1 and 2 of cadherin 11 (rFN/CDH) using a dimethyl-3,3'-dithiobispropionimidate cross-linking method. The surface was characterized by scanning electron microscopy, X-ray photoelectron spectroscopy and protein adsorption and surface density measurements. The material exhibited desirable properties for cell adhesion and proliferation. The effects of the surface on the adhesion and proliferation of human mesenchymal stem cells (hMSC) were investigated using a cell adhesion centrifugal assay and the 3-(4,5-dmethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide method. The data demonstrated that the adhesive capacity and proliferation rate were significantly improved as compared with fibronectin and cadherin positive controls. Moreover, the rFN/CDH bio-inspired ceramic surface also induced osteoblastic differentiation, as evidenced by the higher alkaline phosphatase activity and osteocalcin mRNA expression level of hMSC cultured in osteogenic media for 7-10days. Furthermore, a functional blocking assay with a site-specific antibody against phosphotyrosine 397 (pY397) of focal adhesion kinase revealed that pY397 is involved in adhesion and ossification. These results suggest that the rFN/CDH bio-inspired BCP surface possesses enhanced functionality in adhesion, proliferation and ossification and may be a promising scaffold for tissue engineering.
机译：这项研究已经研究了用新型重组蛋白修饰的生物启发陶瓷表面对表面参数和细胞行为的影响。使用二甲基-3,3'-dithiobispropionimidate交联，用跨纤连蛋白模块III7-10片段和钙粘蛋白11的胞外域1和2（rFN / CDH）的重组蛋白对两相磷酸钙陶瓷的表面进行功能化。方法。通过扫描电子显微镜，X射线光电子能谱以及蛋白质吸附和表面密度测量来表征表面。该材料表现出细胞粘附和增殖的理想特性。使用细胞粘附离心测定法和3-（4,5-dmethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide方法研究了表面对人间充质干细胞（hMSC）粘附和增殖的影响。数据表明，与纤连蛋白和钙黏着蛋白阳性对照相比，粘附能力和增殖速率显着提高。此外，rFN / CDH生物启发的陶瓷表面也诱导了成骨细胞分化，这由成骨培养基中培养7-10天的hMSC的较高碱性磷酸酶活性和骨钙素mRNA表达水平所证明。此外，使用针对粘着斑激酶的磷酸酪氨酸397（pY397）的位点特异性抗体的功能阻断试验显示，pY397参与粘附和骨化。这些结果表明，rFN / CDH生物启发的BCP表面在粘附，增殖和骨化方面具有增强的功能，并且可能是组织工程的有希望的支架。

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