明胶—壳聚糖复合神经营养素3神经支架材料的构建及其生物相容性检测  

Establishment of gelatin-chitosan composite neurotrophins-3 nerve scaffold and its biocompatibility

作　　者：秦丽娜[1] 

机构地区：[1]中山大学中山医学院,广州510080

出　　处：《山东医药》2015年第7期1-4,I0001,共5页Shandong Medical Journal

基　　金：国家自然科学基金资助项目(31200737);广东省医学科研基金立项课题(B2012080)

摘　　要：目的构建明胶—壳聚糖复合神经营养素3神经支架材料,并检测其生物相容性。方法混合明胶、壳聚糖溶液后加入微量神经营养素3,将混浊液注模成型后冷淋干燥制备明胶—壳聚糖复合神经营养素3神经支架材料,利用扫描电子显微镜观察支架材料形态,液体代替法测算孔隙率。提取明胶—壳聚糖复合神经营养素3神经支架材料浸提液,观察其对神经干细胞活性的影响及对全反式维甲酸预诱导的神经干细胞分化的影响,并采用膜片钳技术检测诱导分化前后细胞的电生理特性。结果明胶—壳聚糖复合神经营养素3神经支架材料内径为(267.0±13.8)μm,孔隙率为90.0%。神经干细胞在支架材料上生长良好,在全反式维甲酸诱导下形态向神经元样细胞改变,并初步表现出神经元间突触连接的结构,且诱导分化的神经元样细胞初步具备神经元细胞的电生理特性。结论成功构建了明胶—壳聚糖复合神经营养素3神经支架材料,其与神经干细胞之间的生物相容性良好。Objective To fabricate the gelatin-chitosan composite neurotrophins-3 nerve scaffolds and to determine its biocompatibility .Methods We mixed the gelatin and chitosan solutions , then added a trace of neurotrophin-3.After mak-ing the injection mold , we fabricated the nerve guidance scaffolds by the freeze-drying technique .Then the characteristics of the scaffold were observed by scanning electron microscope morphology , and the scaffold diameter , porosity, etc were de-tected.We extracted the gelatin chitosan composite neurotrophin-3 nerve scaffold material extracts , and then we observed the effects on the growth and differentiation of neural stem cells;the electrophysiological properties of the differentiated cells were observed by the patch-clamp technique .Results The inside diameter of gelatin-chitosan composite neurotrophins-3 nerve scaffold was （267.0 ＆#177;13.8） μm and porosity was 90.0%.Scanning electron microscopy showed that neural stem cells grew well on the scaffold.Under the induction of retinoic acid , neural stem cells differentiated into neuron-like cells, and showed structural of synaptic connections between neurons initially .What′s more, the cells differentiated were detected the neuron-like cell electrophysiological properties by the patch-clamp technique .Conclusion The gelatin-chitosan com-posite neurotrophin-3 nerve scaffolds were successfully constructed , and had good biocompatibility with neural stem cells .

关 键 词：脊髓损伤 支架材料 神经营养素3 维甲酸 神经干细胞 

分 类 号：R318[医药卫生—生物医学工程]