壳聚糖聚电解质复合物再生纤维软骨的实验研究

Experimental study on chitosan-polyelectrolyte complex as a scaffold for cartilage regeneration in vivo

目的:探讨壳聚糖电介质复合物与髁突软骨细胞体内形成纤维软骨情况,以期为工程化软骨再生修复颞下颌关节软骨缺损奠定基础。方法:合成磷酸化壳聚糖电解质复合物(chitosan-polyelectrolyte complex,CS-PEC)制备三维凝胶支架,并与髁突软骨细胞复合培养后植入裸鼠体内,分别于4、8周取出新生物,通过组织学和免疫组化观察新生物的生物学特性。结果:软骨细胞在CS-PEC支架中形态铺展良好。植入体内4周后,支架保持原有结构,可见肥大样软骨细胞出现,免疫组化染色显示中央新软骨形成区Ⅱ胶原为阳性。8周后,支架降解,见新生软骨形成,阿新兰染色显示软骨基质形成,免疫组化染色显示软骨形成区Ⅱ胶原为阳性,周围纤维样细胞中Ⅰ胶原阳性;对照组无新生软骨形成,软骨特异性标志Ⅱ胶原及软骨基质特染均为阴性。结论:CS-PEC复合材料有望成为软骨再生的支架载体。

Objective：To investigate whether chitosan-polyelectrolyte complex （CS-PEC） can be used as scaffold for chondrocyte cuhure and for cartilage regeneration in vivo. Methods：Condylar chondrocytes of fetal mouse were seeded onto the three-dimension gel scaffolds of CS-PEC and cuhured. The cuhured chondrocytes/CS-PEC complex samples were transplanted subcutaneously into nude mice and the CS-PEC scaffold without chondrocyte was used as the control. The animals were sacrificed 4 and 8 weeks after operation respectively. Cartilage formulation was observed by histological and immunohistochemical methods. Results：In the in vitro cuhure the majority of cells attached to the CS-PEC surface and expanded rapidly. 4 v, eeks after transplantation,in the chondrocytes/CS-PEC complex the scaffold maintained mostly the original structure. Hypertrophic chondrocytes appeared in scaffold materials. Collagen Ⅱ was positive in the new cartilage. 8 weeks after transplantation the scaffold degraded almost completely and new cartilage could be ohserved. Collagen Ⅱ and cartilage matrix was positive in the new cartilage and the collagen Ⅰ was positive in the surruunding fihrohlast-like cells. In cnntrol transplants,8 week after transplantation some fibre-like tissue fomted in the circumference, hut there was no new cartilage formation and the collagen Ⅱ and the cartilage matrix was negative. Conclusions：CS-PEC may be used as scaffold for fibre-cartilage regeneration.