钛表面类骨磷灰石上胶原蛋白与羟胆固醇复合生物活性涂层制备研究

Preparation of bioactive composite layers by loading collagen and 20α-hydroxycholesterol on the surface of apatite-coated Titanium substrates

目的探讨在金属钛表面类骨磷灰石上制备胶原蛋白与羟胆固醇复合生物活性涂层的可行性。方法研究于2013年12月在清华大学材料学院进行。采用酸-碱热处理结合仿生矿化沉积的方法在钛表面构建一层均匀的类骨磷灰石,采用物理吸附方法将胶原蛋白和羟胆固醇共同或单一固定在具有类骨磷灰石涂层的钛表面,构建钛表面的复合涂层。结果场发射扫描电镜和X射线衍射分析验证了钛表面的类骨磷灰石;衰减全反射-傅里叶变换红外光谱和X射线光电子能谱定性证明了胶原蛋白被成功固定在具有类骨磷灰石涂层的钛表面上;微二喹啉甲酸法和高效液相色谱法可测定固定在复合涂层上的胶原蛋白和羟胆固醇含量。结论通过酸-碱热处理和仿生矿化沉积方法可在钛表面构建一层均匀的类骨磷灰石,通过物理吸附方法可成功将胶原蛋白和羟胆固醇共同或单一固定在具有类骨磷灰石涂层的钛表面,构建钛表面的复合涂层。

Objective To prepare the composite layers by loading collagen （Col）and 20α-hydroxycholesterol （HC） on the surface of apatite-coated Titanium （Ti） substrates. Methods Apatite was firstly preformed on acid-alkali-treated Ti substrate by immersing in Dulbeceo＇s phosphate-buffered saline （DPBS）solution containing CaC12. Col and HC were absorbed individually or simultaneously onto the surface of the biomimetically formed apatite. Results The field emis- sion scanning electron microscopy （FESEM） and X-ray diffraction （XRD）revealed apatite was formed on the surface of Ti substrates. The qualitative analysis of Col was carried out using attemuated total reflectance Fourier transformed infra- red spectroscopy（ATR-FTIR）and X-ray photoelectron spectroscopy（XPS）. The quantitative analysis of Col and HC was performed by micro bicinchoninic acid （BCA） assays and high preformance liquid chromatography （HPLC）. Conclusion Col and HC are effectively loaded onto Ti substrates, initially coated with biomimetie apatite, by simply dropping solu- tions containing either Col or HC.