SHH修饰聚多巴胺涂层纤维蛋白支架对大鼠神经干细胞的影响  

Polydopamine-mediated surface modification of fibrin scaffold with SHH coating for rat neural stem cell engineering

作　　者：崔学文[1] 陆浩 吕德民 沈元昊 杨开元 曹苏成 杨文静[2] 张志坚[2] Cui Xuewen;Lu Hao;Lv Demin;Shen Yuanhao;Yang Kaiyuan;Cao Suchen;Yang Wenjing;Zhang Zhijian(Department of Orthopedics,Jiangbin Hospital Affiliated,Jiangsu University;School of Medicine,Jiangsu University,Zhenjiang 212013,China)

机构地区：[1]江苏大学附属医院骨科 [2]江苏大学医学院,镇江212013

出　　处：《神经解剖学杂志》2020年第1期15-22,共8页Chinese Journal of Neuroanatomy

基　　金：国家自然基金(51403086);江苏省自然科学基金(BK20140544);江苏大学青年英才培育计划(4111280006)。

摘　　要：目的:构建音猬因子(SHH)修饰的聚多巴胺(PD)涂层纤维蛋白胶(FG),并探究该支架对大鼠神经干细胞(NSCs)的生长及分化的影响。方法:按FG材料表面涂层情况,将实验分为3组:FG表面未处理组(control)、FG表面进行多巴胺涂层组(PD)和FG经多巴胺处理后表面涂有SHH组(PD-SHH)。采用扫描电子显微镜分别观察支架微观结构,用ELISA测量PD-SHH组SHH。分离并培养大鼠神经干细胞,并将神经干细胞分别与该3种支架复合培养,免疫荧光及Western Blot检测各组细胞中胶质纤维酸性蛋白(GFAP)、生长再生相关蛋白(GAP43)和髓鞘碱性蛋白(MBP)的表达。结果:ELISA结果显示PD-SHH缓释支架可以持续释放SHH达到19 d;扫描电子显微镜可见纤维蛋白胶经冷冻干燥后呈海绵型网状结构,多巴胺及SHH微粒粘附在网状结构上;免疫荧光及Western Blot结果显示:PD-SHH组细胞高表达GAP43和MBP等神经细胞相关蛋白而低表达GFAP(P<0.05)。结论:制备的聚多巴胺涂层纤维蛋白支架具有良好的SHH缓释效果,该复合支架对大鼠NSCs分化有明显的促进作用。Objective:To construct a polydopamine-mediated surface modification of fibrin glue scaffold adhering sonic hedeghog and explore its effect on the differentiation of rat neural stem cell(NSCs).Methods:According to thematerials coated on the surface of FG,the protein scaffold were divided into three groups:Control group(without any coating),PD group(coating with polydopamine),PD-SHH group(coating with polydopamine and SHH).The microstructure of the scaffold was observed by scanning electron microscopy and the sustained release effect of SHH in the PDSHH group was measured by ELISA.Neurospheres were cultured from fetal mice brain and then characterized,and the rat neural stem cells were cultured with the three scaffolds respectively.After 14 days,immunohistochemistry and Western Blot were used to investigate the espression of Nestin,GAP43,MBP,and GFAP.Results:PD-SHH has a good performance of sustained-release of SHH;Scanning electron microscopy(SEM)shows:fibrin glue scaffold has a spongelike network structure after lyophilization,Dopamine and SHH particles adhere to the network structurally.Immunofluorescence and Western Blot results showed that the expression of GAP43,MBP in the PD-SHH group was higher than that in Control and PD groups(P<0.05),while GFAP was lowly expressed(P<0.05).Conclusion:PD-SHH scaffold has a good sustained release effect of SHH and promoted the differentiation of rat NSCs significantly.

关 键 词：纤维蛋白支架 多巴胺 音猬因子 神经干细胞 大鼠 

分 类 号：R318.08[医药卫生—生物医学工程]