以PGA为三维支架同种异体工程化软骨的构建

FABRICATION OF TISSUE ENGINEERED CARTILAGE USING PGA AS SCAFFOLD SEEDED WITH ALLOGENIC CHONDROCYTES

为探讨以聚羟基乙酸 (PGA)为三维支架材料的同种异体软骨细胞构建组织工程化软骨的能力 ,采用胶原酶消化方法分离乳兔肋软骨获取种子细胞 ,收集体外培养传 2～ 3代的软骨细胞 ,接种于经多聚赖氨酸处理的PGA支架材料上 ,将细胞 材料复合物种植在成兔皮下 ,一定时间取材 ,对获得的同种异体工程化软骨进行组织学评价。结果显示 ,软骨细胞体外培养 1天后 ,在PGA支架上分布均匀 ;培养 1周左右 ,PGA纤维间有呈蜘蛛网状基质产生。种植复合物 4周取材可见PGA纤维未完全降解 ,软骨细胞不成熟 ,周边存在一定程度炎细胞浸润 ;8周取材 ,PGA纤维消失 ,软骨基本成熟 ,炎细胞浸润不明显 ,软骨基质含量丰富 ,分布与正常肋软骨组织学特征相似。提示以PGA为支架材料同种异体软骨细胞在有免疫力的动物体内可形成工程化软骨 。

To investigate the formation of tissue engineered cartilage using polyglycolic acid (PGA) as three dimensional scaffold seeded with allogenic chondrocytes. First, chondrocytes were harvested from rib cartilage of one week old infant rabbits by collagenase digestion.The isolated cells were cultured in vitro for 2～3 generations, and then the cell suspension was concentrated. Next, the suspension was seeded onto PGA scaffolds soaked with poly l lysine. Then, scaffold chondrocyte constructs were transplanted into subcutaneous tissue of adult rabbits. Finally, the chondrogenesis was studied histologically at various periods. The results showed that chondrocytes were well distributed on the PGA scaffolds in the period of culture in vitro, and could secrete matrix around one week after seeding. 4 weeks after transplantation, the immature cartilage was regenerated in recipient animals. But some inflammatory cells surrounding the new cartilage and some PGA fibers were observed. 8 weeks after transplantation, mature cartilage with abundant matrix was obtained. PGA fibers were not found. This study proves that it is possible to regenerate new cartilage in animals possessing immune function by seeding allogenic chondrocytes onto PGA scaffold. Rejectionreaction is not strong.