【摘要】 目的制备一种可缓慢释放神经生长因子(NGF)的的高分子复合材料,用于神经组织工程的研究。方法采用溶剂挥发的方式,将NGF与聚羟基脂肪酸酯(PHA)在有机溶剂中充分混合,再将其倒入圆形玻璃器皿中,形成一个表面和内部都含有NGF的PHA膜,并将纯的PHA膜和浸泡干燥的PHA膜(粘附NGF的PHA膜)作为对照。将上述三种膜分别进行微观形貌(扫描电子显微镜)、水接触角和NGF体外释放的检测。最后,在连续培养6 d后,观察三种膜对PC12细胞的神经分化影响。结果相对于两个对照组,负载NGF的PHA膜表面形貌更起伏粗糙,有明显的蛋白粘附,也具有最佳的亲水性。同时,负载NGF的PHA膜能连续6天释放出稳定的且有活性的NGF,浓度大于100 ng/m L。负载NGF的PHA膜上的PC12细胞分化明显,在培养第3天时出现神经样细胞的分化表型,长出了轴突,而第6天邻近的几个PC12细胞的轴突相互接触并形成类似神经网结构;其他对照组没有明显现象。结论成功利用生物材料PHA制备出一种具有NGF缓释功能的生物高分子膜片,并可诱导PC12细胞向神经细胞表型分化。更多还原

【Abstract】 Objective To study the preparation of polyhydroxyalkanoate(PHA) film with nerve growth factor(NGF) slow release and its effect on neural differentiation of PC12 cells.Methods NGF and PHA were mixed thoroughly in the organic solvent through solvent evaporation method.Then the solvent was poured into a round glass container to form a PHA film containing NGF inside and outside.The pure PHA film and the soaked dry PHA film(PHA film attached NGF) were regarded as the control groups.The three kinds of films were characterized by microstructure(SEM),water contact angle measurement and NGF release assay respectively.After 6 consecutive days of culturing,the effects of three kinds of films on neuronal differentiation of PC12 cells were observed.Results Compared by the twocontrol groups,the surface of PHA films loaded with NGF appeared rougher,and had obvious protein adhesives and the best hydrophilia.Meanwhile,the PHA films loaded with NGF could release the active NGF whose concentrations exceeded 100 ng/ml for 6 consecutive days steadily.The PHA films loaded with NGF had the obvious differentiation.On the third day of culturing,the differentiation phenotype of neuron-like cells appeared and the axons grew;on the sixth day of culturing,the axons of neighboring PC12 cells contacted with each other and formed a structure of resembling neural network.While,the control groups had no obvious differentiation phenomena.Conclusion In this study,a biopolymer biomembrane with a slow-release function of NGF was successfully prepared and the PC12 cells were induced to differentiate into neuronal cells.更多还原