摘要:
生物材料表面的粗糙度是影响细胞行为的重要因素之一。为了调控丝蛋白生物材料表面的粗糙度, 并评价材料表面粗糙度对细胞生长行为的影响, 首先, 通过湿化学共沉淀法, 以柞蚕丝胶(AS)溶液为模板, 诱导了羟基磷灰石(HAp)晶体成核, 进而调控了AS膜表面的粗糙度。然后, 采用SEM、粗糙仪、FTIR及EDX等对HAp/AS复合膜表面形貌、粗糙度及成分进行了表征。最后, 通过SEM和CellTiter 96® AQueous单溶液细胞增殖检测试剂盒(MTS)检测了骨髓间充质干细胞(BMSCs)在HAp/AS复合膜表面的形貌及增殖率。结果表明:纯AS膜的表面粗糙度为0.15 μm, 矿化1、8及24 h后, 表面粗糙度分别为0.38、0.46和1.20 μm;矿化24 h后, 在HAp/AS复合膜表面可观察到直径为30~80 nm的球状复合物, 生成的矿化物为HAp;HAp/AS复合膜具有良好的细胞相容性, 表面粗糙度为1.20 μm的复合膜能够显著促进BMSCs的增殖, 粗糙度对BMSCs在HAp/AS复合膜表面的粘附和形貌有着重要的影响。因此, 可通过矿化的方法在生物大分子表面诱导HAp晶体的成核与生长, 从而调控材料的表面粗糙度, 研究材料界面上的细胞行为。
Abstract:
The surface roughness of biomaterial is one of the important factors which affect the cell behavior. In order to regulate the surface roughness of silk protein biomaterials and evaluate the effects of the surface roughness of material on cell growth behavior, Antheraea pernyi silk sericin (AS) solution was used as the template to induce the nucleation of hydroxyapatite (HAp) crystals by wet chemical coprecipitation method firstly, and therefore regulated the surface roughness of AS films. Then, the surface morphology, roughness and elements of HAp/AS composite films were characterized with SEM, roughometer, FTIR and EDX. Finally, the cell morphology and proliferation rate of bone mesenchymal stem cells (BMSCs) on the surface of HAp/AS composite films were detected with SEM and CellTiter 96® AQueous one solution cell proliferation assay reagent (MTS). The results show that the surface roughness of pure AS film is 0.15 μm. After mineralization for 1, 8 and 24 h, the surface roughness is 0.38, 0.46 and 1.20 μm, respectively. After mineralization of 24 h, some lobular compounds whose diameter is 30-80 nm can be observed on the surface of AS/HAp composite films. The resultant mineralized compound is HAp. HAp/AS films have good biocompatibility and the composite film whose surface roughness is 1.20 μm can promote the proliferation of BMSCs significantly. The roughness has important effect on attachment and morphology of BMSCs on the surfaces of HAp/AS films. Thus, the nucleation and growth of HAp crystal can be induced on the surfaces of biomacromolecules by the method of mineralization, there by regulate the surface roughness of materials, and investigate the cell behavior on the interface of materials.
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