Institut für Anorganische Chemie, Leibniz Universität Hannover, Hannover, Germany.
Acta Biomater. 2011 Apr;7(4):1772-9. doi: 10.1016/j.actbio.2010.12.028. Epub 2010 Dec 25.
Due to its ability to induce de novo bone formation the differentiation factor bone morphogenetic protein 2 (BMP2) is often used to enhance the integration of bone implants. With the aim of reducing possible high dose side-effects and to lower the costs, in order to produce affordable implants, we developed a simple and fast method for the immobilization of BMP2 on silica-based surfaces using silane linkers which carry amino or epoxy functions. We put an especial emphasis on the influence of the nanoscale surface topography of the silica layer. Therefore, we chose glass (for control experiments) and Bioverit® II (as a typical implant base material) as support materials and coated these substrates with unstructured or nanoporous amorphous silica layers for comparison. Immobilized BMP2 was quantified by two different methods: by ELISA and by a cell-based assay for active BMP2. These tests probe for immunologically and biologically active BMP2, respectively. The results show that the amino functionalization is better suited for immobilizing the protein. Strikingly, a considerably higher amount of BMP2 could be immobilized on coated Bioverit® II surfaces compared with coated glass substrates, which was presumably due to the macroscopic roughness of the Bioverit® II substrates. In addition, it was found that the nanoporous silica coatings on Bioverit® II substrates were able to bind more BMP2 than the unstructured ones.
由于其诱导新骨形成的能力,分化因子骨形态发生蛋白 2(BMP2)常被用于增强骨植入物的整合。为了降低可能的高剂量副作用并降低成本,以生产出负担得起的植入物,我们开发了一种使用带有氨基或环氧功能的硅烷连接子将 BMP2固定在基于硅的表面上的简单快速方法。我们特别强调了硅层纳米级表面形貌的影响。因此,我们选择玻璃(用于对照实验)和 Bioverit® II(作为典型的植入物基础材料)作为支撑材料,并将这些基底涂覆无定形的非孔或纳米多孔硅层进行比较。通过两种不同的方法定量测定固定化的 BMP2:通过 ELISA 和基于细胞的活性 BMP2 测定。这些测试分别探测免疫和生物活性 BMP2。结果表明,氨基功能化更适合固定蛋白质。引人注目的是,与涂覆玻璃基底相比,涂覆的 Bioverit® II 表面可以固定更多的 BMP2,这可能是由于 Bioverit® II 基底的宏观粗糙度所致。此外,还发现 Bioverit® II 基底上的纳米多孔硅涂层比无定形涂层能够结合更多的 BMP2。