神经干细胞与透明质酸水凝胶材料生物相容性的研究

A study on the biocompatibility of neural stem cells with a newly created biomaterial of hyaluronic acid based scaffold

目的:评价神经干细胞与改性透明质酸水凝胶支架新材料的生物相容性,研究该透明质酸水凝胶支架作为中枢神经组织工程载体材料的可行性,为用组织工程及干细胞技术治疗中枢神经系统损伤提供基础。方法:以冷冻干燥法制备透明质酸水凝胶材料支架,通过化学接枝法将抗Nogo受体抗体(Anti-NogoR)和多聚赖氨酸(poly-l-lysine,PLL)分子接枝到水凝胶上对其改性,制成新的支架材料。体外培养胚胎13.5d大鼠前脑泡神经干细胞.将神经干细胞与生物支架共培养,通过扫描电镜观察透明质酸水凝胶的内部结构及神经干细胞在支架材料上的粘附与生长情况,通过细胞免疫组织化学技术观察神经干细胞在透明质酸水凝胶材料上的存活与分化情况。结果:制备的透明质酸水凝胶材料具有疏松的三维多孔结构,神经干细胞在支架材料上能够粘附并且有突起长出,生长良好。神经干细胞在支架材料上能够分化。结论:神经干细胞与经过改性的透明质酸水凝胶新材料有很好的生物相容性,能够在材料上存活分化。该新透明质酸水凝胶材料有望作为修复中枢神经损伤的组织工程载体。

Objective：To evaluate the biocompatibility of neural stem cells（NSCs）with a newly created biomaterial hydrogel and investigate the feasibility of hyaluronic acid（HA）scaffold for the tissue engineering of central nervous system（CNS）,in order to provide the basis for the treatment of CNS injury by the use of tissue engineering and stem cell technologies.Methods：A three dimensional HA based hydrogel,as a biodegradable tissue engineering scaffold to mimic the structure and components of the extracellular matrix（ECM）in the brain,was made by a technique of frozen-lyophilization.The molecules of poly-l-lysine（PLL）were immobilized on the backbone of the hydrogel for improving cell adherence and compatibility with brain tissue.Meanwhile,an antibody releasing system was developed by covalently attaching the anti-NogoR to the HA.The NSCs were isolated from the forebrain of rat embryo（E 13.5 d）and cultured on the hydrogel scaffold.The microstructure of the material and adhesion of the NSCs were observed under scanning electron microscopy（SEM）.Growth and differentiation of the NSCs on the scaffold were determinated by immunohistochemistry.Results：A poriferous three-dimenssion structure of the HA scaffold was observed with SEM and it was highly bionic.The NSCs have good adhesion with the scaffolds and could grow neurite,so that they were survived well in the scaffolds.The NSCs can also differentiate in the HA hydrogel in vitro.Conclusion：Our newly created biomaterial has a good biocompatibility with cultured NSCs and it might be useful for repairing the injury in the CNS,as a feasible approach of neural tissue engineering.