Biomaterials, Biomechanics and Tissue Engineering Group, Department of Materials Science and Metallurgical Engineering, Technical University of Catalonia (UPC), ETSEIB , Diagonal 647, 08028 Barcelona, Spain.
Centre for Research in NanoEngineering (CRnE)-UPC , c/Pascual i Vila 15, 08028 Barcelona, Spain.
ACS Appl Mater Interfaces. 2016 Feb 3;8(4):2517-25. doi: 10.1021/acsami.5b09576. Epub 2016 Jan 20.
Since metallic biomaterials used for bone replacement possess low bioactivity, the use of cell adhesive moieties is a common strategy to improve cellular response onto these surfaces. In recent years, the use of recombinant proteins has emerged as an alternative to native proteins and short peptides owing to the fact that they retain the biological potency of native proteins, while improving their stability. In the present study, we investigated the biological effect of two different recombinant fragments of fibronectin, spanning the 8-10th and 12-14th type III repeats, covalently attached to a new TiNbHf alloy using APTES silanization. The fragments were studied separately and mixed at different concentrations and compared to a linear RGD, a cyclic RGD and the full-length fibronectin protein. Cell culture studies using rat mesenchymal stem cells demonstrated that low to medium concentrations (30% and 50%) of type III 8-10th fragment mixed with type III 12-14th fragment stimulated cell spreading and proliferation compared to RGD peptides and the fragments separately. On the other hand, type III 12-14th fragment alone or mixed at low volume percentages ≤50% with type III 8-10th fragment increased alkaline phosphatase levels compared to the other molecules. These results are significant for the understanding of the role of fibronectin recombinant fragments in cell responses and thus to design bioactive coatings for biomedical applications.
由于用于骨置换的金属生物材料生物活性低，因此使用细胞黏附部分是改善这些表面细胞反应的常用策略。近年来，由于重组蛋白保留了天然蛋白的生物学效力，同时提高了其稳定性，因此已成为天然蛋白和短肽的替代物。在本研究中，我们使用 APTES 硅烷化法研究了两种不同的纤连蛋白重组片段对新 TiNbHf 合金的生物效应，这两种片段分别跨越 8-10 型和 12-14 型 III 型重复序列。这些片段分别进行了研究，并以不同的浓度混合，与线性 RGD、环状 RGD 和全长纤连蛋白蛋白进行了比较。使用大鼠间充质干细胞的细胞培养研究表明，与 RGD 肽和片段相比，低浓度（30%和 50%）的 III 型 8-10 片段与 III 型 12-14 片段混合可刺激细胞铺展和增殖。另一方面，单独的 III 型 12-14 片段或与 III 型 8-10 片段以低体积百分比（≤50%）混合可提高碱性磷酸酶水平与其他分子相比。这些结果对于理解纤连蛋白重组片段在细胞反应中的作用以及设计用于生物医学应用的生物活性涂层具有重要意义。
