三种支架材料与软骨细胞复合构建组织工程化软骨的研究

Study on construction of tissue-engineered articular cartilage by three kinds of scaffolds combined with chondrocytes

目的 研究细胞、材料及孔径在诱导关节软骨发育过程中的作用。方法 采用细胞与支架的三维培养和体内植入技术。结果 种入支架的原代软骨细胞增殖差 ,未形成软骨组织。而第 3代软骨细胞生长良好 ,最终发育成软骨组织。孔径为 2 5～ 1 0 0 μm的胶原海绵捕捉的细胞量明显多于孔径为 1 0 0～ 550 μm的明胶海绵和PDLLA泡沫。裸鼠皮下植入 1 6周时 3种支架 细胞复合体比较 ,胶原海绵 细胞复合体中的支架材料降解最彻底 ,软骨组织发育最好。结论 第 3代软骨细胞细胞增殖优于原代细胞 ;2 5～ 1 0 0 μm的小孔径支架对细胞的吸附量优于 1 0 0～ 550 μm的大孔径支架 ;

Objective To investigate the roles of chondrocytes,scaffolds and pore size in the chondrogenesis of tissue engineered cartilage.Methods The techniques of three dimentional cell culture and implantation were used.Results The primary chondrocytes seeded to any scaffold were not proliferated and did not have any neocartilage formation.In contrary,the chondrocytes of the third passage seeded into the scaffolds had an excellent proliferation and eventually regenerated neocartilage.The numbers of chondrocytes seeded into collage sponges whose pore diameters were between 25 and 100 μm were much more than those of the gelatin sponge and PDLLA foam,whose pore diameters between 100 and 550?μm.In comparsion among the constructs having been cultured in nude mice for 16 weeks,the neocartilages from the collage chondrocyte constructs,in which scaffolds degraded most thoroughly,developed best.Conclusions On the basis of the cell proliferation the third passage chondrocytes were superior to the primary chondrocytes.On the basis of the number of chondrocytes attaching the scaffolds the small pore scaffolds (25 to 100 μm) possess more advantages than the large pore scaffolds (100 to 550?μm).On the basis of the quality of the cartilage formation of the contrusts,the thorough degradation of the scafflods in constructs is the essential prerequisite for the excellent quality of neocartilage formation.