Departamento de Química Inorgánica y Bioinorgánica, Facultad de Farmacia, Universidad Complutense de Madrid, E-28040 Madrid, Spain.
Acta Biomater. 2010 Mar;6(3):743-9. doi: 10.1016/j.actbio.2009.09.004. Epub 2009 Sep 12.
Silicon-doped hydroxyapatite has been functionalized with biotin molecules as a new methodology for the attachment of proteins, peptides or growth factors through the formation of avidin-biotin complex in this material. Bioceramic biotinylation has been performed by esterification reaction between the OH groups of hydroxyapatite and COOH groups of biotin molecules. Several parameters of the biotinylation, such as the addition of N,N'-dicyclohexylcarbodiimide (DCC), the biotin/bioceramic molar ratio and the activation time, have been studied in order to improve both the amount of anchored biotin on the bioceramic surface and its bond strength. The grafting of biotin on a silicon-doped hydroxyapatite surface was determined using (13)C nuclear magnetic resonance, Fourier transform infrared spectroscopy and elemental analyses. The results show that the addition of DCC significantly increases both the amount of biotin grafted and the bond strength, because the major part is through covalent bonding. Lixiviation studies in simulated body fluid (SBF) at 37 degrees C have confirmed such results, showing that the retention grade after 7 days in SBF was of ca. 63%. Fluorescein isothiocyanate-avidin complexation has been performed on three-dimensional (3-D) scaffolds prepared by a rapid-prototyping technique. Confocal microscopy studies show a homogeneous distribution with a higher incorporation rate of the protein over the entire external surface of the biotinylated 3-D scaffold.
硅掺杂羟基磷灰石经生物素分子功能化，通过在该材料中形成亲和素-生物素复合物，作为一种新方法用于蛋白质、肽或生长因子的附着。生物陶瓷的生物素化是通过羟基磷灰石的 OH 基团与生物素分子的 COOH 基团之间的酯化反应来实现的。为了提高生物陶瓷表面固定化生物素的数量及其键合强度，研究了生物素化的几个参数，如 N,N'-二环己基碳二亚胺（DCC）的添加、生物素/生物陶瓷摩尔比和活化时间。（13）C 核磁共振、傅里叶变换红外光谱和元素分析表明，生物素接枝到硅掺杂羟基磷灰石表面。结果表明，DCC 的添加显著增加了接枝的生物素的数量和键合强度，因为大部分是通过共价键结合的。在 37°C 的模拟体液（SBF）中的浸出研究证实了这一结果，表明在 SBF 中 7 天后的保留率约为 63%。荧光素异硫氰酸酯-亲和素复合物已在通过快速成型技术制备的三维（3-D）支架上进行。共焦显微镜研究表明，在整个生物素化 3-D 支架的外表面上，蛋白质的分布均匀，且掺入率更高。
