应用高浓度Ⅱ型胶原蛋白构建组织工程软骨支架

Construction of scaffold for treatment of articular cartilage defect using high-concentration collagen Ⅱ

目的构建有利于种子细胞黏附、增殖和分化的软骨组织工程海绵支架。方法依据天然软骨主要成分,提取透明软骨Ⅱ型胶原蛋白,将提取的胶原蛋白通过高速离心进行浓缩,真空冷冻干燥制得海绵支架材料,应用化学交联剂EDC/NHS进行化学交联,对支架采用组织化学染色、体外降解实验、扫描电镜和接种细胞静态培养的方法进行相关检测。结果支架具有良好的孔隙结构,孔径40～130μm,孔隙率在90%以上,细胞在支架上增殖、分化良好。结论构建的高浓度Ⅱ型胶原蛋白海绵支架具有良好的生物相容性,有较好的应用前景。

Objective To construct a sponge scaffold for cell adhesion,proliferation and differentiation in cartilage tissue engineering.Methods Collagen Ⅱ was extracted from hyaline cartilage,and was then subjected to high-speed centrifugation and vacuum freeze-drying to obtain a sponge scaffold.The sponge scaffold was soaked in 1-ethyl-3-（3-dimethylaminopropyl） carbodiimide（EDC）/N-hydroxysuccinimide（NHS） for cross-linking.The sponge scaffold was evaluated by scanning electron microscopy,immunohistochemistry,degradation test and cell adhesion and culture.Results The sponge scaffold had a good pore structure,with pore sizes of 40-130 μm and porosity above 90%.Good cell proliferation and differentiation were found on the scaffold.Conclusion The sponge scaffold constructed from high-concentration collagenⅡ has excellent biocompatibility,and shows great promise in the treatment of articular cartilage defect.