利用胶原和聚乙烯醇构建组织工程软骨支架

Construction of scaffold for cartilage tissue engineering using collagen and polyvinyl alcohol

目的依据仿生学原理,应用Ⅱ型胶原蛋白(type-Ⅱcollagen,CoⅡ)、硫酸软骨素(chondroitin sulfate,CS)、透明质酸(hyaluronan,HA)和聚乙烯酸(polyvinyl alcohol,PA)来制备组织工程支架材料。方法依据人体正常关节软骨细胞外基质主要成分Ⅱ型胶原、硫酸软骨素和透明质酸;添加高分子聚合物PVA来提高支架强度,应用冷冻干燥等技术制备复合支架,接种细胞体外静态培养7d,通过力学测试、扫描电镜和组织染色等方法对支架进行检测。结果支架孔隙均匀,孔隙率为93%～95%,CoⅡ-CS-HA-PVA复合支架最大张力约为1.19N,单纯CoⅡ-CS-HA复合支架约为0.55N,差异有统计学意义(t=4.985,P=0.002),细胞在支架上均匀分布,活力旺盛。结论 CoⅡ-CS-HA-PVA复合支架具备良好的生物相容性,可为种子细胞增殖分化提供一个适宜的环境,有很好的临床应用前景。

Objective To construct the scaffold for cartilage tissue engineering according to the bionics principles. Methods 3-dimensional scaffolds were prepared with type-Ⅱ collagen （CoⅡ）,hyaluronan （HA） and chondroitin sulfate （CS） using the freeze-dry technology. Polyvinyl alcohol （PVA） was added to enhance their mechanical strength. The scaffolds were evaluated by SME,mechanical test and immunohistochemistry. Then 293 cells were seeded in scaffolds and cultured for 7 d in vitro. Data were analyzed by SPSS13.0. Results The pores of scaffold were uniform with a porosity rate of 93%-95%. The maximum tension of CoⅡ-CS-HA-PVA scaffold and CoⅡ-CS-HA scaffold was 1.19 N and 0.55 N,respectively （t=4.985,P=0.002）. The distribution of cells was even in scaffolds. Conclusion The CoⅡ-CS-HA-PVA scaffold can provide a suitable environment for cell proliferation and differentiation,thus having a very good prospect in clinical application.