接枝Nogo-A抗体和多聚赖氨酸的透明质酸水凝胶支架的制备及其与培养海马神经元相容性的研究

Fabrication of hyaluronic acid hydrogels modified with Nogo-A receptor antibody and poly-L-lysineand its biocompability with hippocampal neurons

为探讨同时接枝Nogo-A受体(NgR)的抗体和多聚赖氨酸(PLL)的透明质酸(HA)水凝胶用于中枢神经系统损伤修复的可行性,本研究在碳二亚胺盐酸盐的介导下用己二酸二酰肼交联的方法制备HA水凝胶,并接枝PLL和NgR抗体。取新生大鼠海马神经元接种于水凝胶支架材料上,分为HA-NgR抗体-PLL水凝胶组和纯HA水凝胶组。培养7d后,用吖啶橙(AO)染色、抗神经丝蛋白(NF)和胶质纤维酸性蛋白(GFAP)免疫荧光染色和扫描电镜观察两组细胞的生长情况。结果显示:纯HA水凝胶不利于神经细胞在材料上粘附和突起生长;HA-NgR抗体-PLL水凝胶可以显著增加海马神经元的粘附数量、促进神经元突起形成,同时也能使星形胶质细胞在材料表面生长。上述结果提示:接枝NgR抗体和PLL的HA水凝胶与海马神经元相容性良好,为中枢神经系统损伤修复提供了一种较理想的支架材料。

To explore the potential application of hyaluronie acid （HA） hydrogels modified with Nogo-A receptor NgR antibody and poly- L-lysine （PLL） in repairing the central nervous system injury, HA hydrogels modified with NgR antibody and PLL were fabricated by using ethyl N ,N-dimethylaminopropyl earbodiimide （EDC） as an activating agent, and crosslinked by Adipie dihydrazides （ADH）. Then, new- born rat hippocampal neurons were planted respectively on pure HA hydrogels or HA hydrogels modified with NgR antibody and PLL. After 7 days co-incubation, the growth of hippoeampal neurons was estimated by the techniques of acridine orange （AO） staining, anti-neurofil- ament （ NF ） , anti-glial fibrillary acidic protein （ GFAP ） immunofluoreseent histoehewical staining and scanning electron microscopy （SEM）. The＇results showed that HA hydrogels modified with NgR antibody and PLL could significantly enhance the attachment of primary hippocampal neurons in number and neurite outgrowth compared with unmodified HA hydrogels. Additionally, some astrocytes were observed to grow on the surface of the modified HA hydrogels. The present result suggest that the HA hydrogel modified with NgR antibody and PLL has a good biocompatibility for the attachment and neurite outgrowth of hippocampal neurons, which can facilitate it to become a useful scaffold for repairing central nervous system injury.