以脱细胞牛软骨基质为支架体外构建组织工程软骨的实验研究

Acellular Cartilaginous Matrix as A Scaffold for Cartilage Reconstruction in vitro

目的以脱细胞牛软骨基质(acellular cartilaginous matrix,ACM)作支架体外构建组织工程软骨,了解其作为软骨组织工程支架的可行性。方法 2003年1月-2005年12月联合应用冻干-反复冻融-酶消化法对牛软骨基质行脱细胞处理。将体外培养扩增的2～5代兔软骨细胞接种在材料上,体外培养3周,观察软骨细胞在支架材料上的生长分布情况。结果软骨细胞在制备的ACM上可较好地黏附生长,并且分泌大量Ⅱ型胶原和葡萄糖胺聚糖;但软骨细胞不能长入ACM内部,只能在表层生长,少量软骨细胞分布在ACM孔隙中。结论 ACM支架材料具有良好的细胞相容性和活性,并且能促进软骨细胞增殖和维持软骨细胞表型。

Objective To investigate the feasibility of applying the acellular cartilaginous matrix（ACM）as the scaffold in constructing tissue engineering cartilage in vitro.Methods We processed the bovine cartilaginous matrix by combination of lyophilization（freeze dry）and repeated freeze-thaw （12cycles）and enzyme digestion in order to remove the cell component.The 2-5passage chondrocytes were seeded onto the ACM,and had been cultured for 3 weeks.The cell growth,attachment and distribution were detected by histochemical stain,immunohistochemical stain,and scan electron microscope.Results The chondrocytes could adhere and grow well on the matrix surface,and synthesize plenty of glycosaminoglycanand typeⅡcollagen.However,the chondrocytes grew only on the surface and superficial layer of the scaffold,and they did not move into the inner part of the scaffold.Conclusion ACM has good cellular compatibility without cytotoxicity and provide temporary substrate to which these anchorage-dependent cells can adhere,and stimulate the chondrocytes anchored on the scaffold to proliferate and keep differentiated phenotype.