定向多通道纤维蛋白支架对EMSCs迁移分化的影响  

The effect of fibrin scaffold with oriented multichannels on the migration and differentiation of EMSCs

作　　者：杨文静[1] 沈元昊 杨开元 陆浩 崔学文[2] 张志坚[1] 陈谦[1] Yang Wenjing;Shen Yuanhao;Yang Kaiyuan;Lu Hao;Cui Xuewen;Zhang Zhijian;Chen Qian(School of Medicine,Affiliated Hospital,Jiangsu University,Zhenjiang 212013,China;Department of Orthopedics,Affiliated Hospital,Jiangsu University,Zhenjiang 212013,China)

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

出　　处：《神经解剖学杂志》2018年第6期658-664,共7页Chinese Journal of Neuroanatomy

基　　金：国家自然基金(51403086,81501077);江苏省自然科学基金(BK20140544);江苏大学高级专业人才科研启动基金(13JDG089)

摘　　要：目的:构建具有定向多通道的纤维蛋白支架,并探究该支架对外胚层间充质干细胞(ecto-mesenchymal stem cells,EMSCs)的生长行为及向神经分化的影响,为其用于脊髓损伤修复提供新思路。方法:通过人工造孔和冷冻干燥技术构建具有定向多通道的纤维蛋白支架,扫描电子显微镜(scanning electron microscope,SEM)观察支架内微观结构。大鼠鼻粘膜来源的EMSCs与该支架复合培养7d,倒置荧光显微镜和SEM评估该支架对EMSCs迁移的引导效果,并通过免疫荧光染色技术鉴定其对EMSCs向神经诱导分化的程度。结果:(1)SEM显示支架横截面上有若干通道空洞,支架纵截面显示通道的走向基本与支架长轴一致;(2)EMSCs粘附在定向通道的孔壁上,且沿着支架纵轴的方向延伸;(3)免疫荧光染色显示神经相关蛋白阳性。结论:EMSCs在定向多通道纤维蛋白支架上能够正常生长,且支架的定向多通道对其生长具有引导作用,利于EMSCs的粘附、迁移和向神经方向分化。Objective: To prepare fibrin scaffold with oriented multichannels,and then,investigate its effect on the migration behavior and neuronal differentiation of ecto-mesenchymal stem cells ( EMSCs) .This work will offer new hope to repair spinal cord injuries.Methods: The fibrin scaffold with oriented multichannels was constructed with artificial pore-forming and freezing techniques,and its morphology was observed with scanning electron microscope ( SEM) .After that,the fibrin scaffold was co-cultured with EMSCs derived from rat nasal mucosa for 7 days,and its effect on guiding on the migration of EMSCs was evaluated with inverted fluorescence microscope and SEM.Finally,immunefluorescent staining technology was employed to determine the differentiation s of EMSCs towards neuron-like cells.Results: ( 1) SEM showed fibrin scaffold with oriented multichannels was obtained,and the multichannels were paralleled with the long axis of the fibrin scaffold.( 2) EMSCs grasped the wall and extended along the directions of the oriented multichannels.( 3) Nerve-related proteins were expressed positively by using immunofluorescent staining.Conclusion: The morphology of EMSCs seeded in fibrin scaffold was normal and the oriented multichannels of fibrin scaffold could guide the adhesion,migration and neuronal differentiation of EMSCs.

关 键 词：定向多通道 纤维蛋白支架 EMSCs 脊髓损伤 

分 类 号：R651.2[医药卫生—外科学]