永生化软骨细胞为基础的工程化软骨修复软骨缺损的实验研究

Experimental study on repair of articular cartilage defects by engineered cartilage based on immortalized chondrocytes

目的 探讨用永生化软骨细胞作为种子细胞修复羊关节软骨缺损的可行性。方法 把人端粒酶催化亚基(hTERT)转染羊关节软骨细胞,筛选阳性克隆并通过旋转生物反应器-微载体技术进行大量扩增;将扩增的永生化软骨细胞与β-磷酸三钙复合并在体外培育后,植入羊前肢肱骨头关节面软骨缺损处;3和6个月取材,进行形态学和免疫组织化学评价。结果 实验组,术后6个月,缺损区被新生的软骨组织所充填,Ⅱ型胶原染色呈强阳性,与对照组差别有显著性差异(P<0.01)。在材料组,缺损区边缘可见部分新生软骨组织;空白对照组,缺损区新生软骨组织形成。结论 永生化软骨细胞在软骨组织工程中具有潜在的应用前景。

Objective To investigate the feasibility of repair of goat articular cartilage defects by tissue engineered cartilage based on immortalized chondrocyte Methods The hTERT gene was introduction into chondrocytes of goat and the positive clones were selected to culture and subculture with rotary cell culture system (RCCS)-microcarrier technique. Immortalized chondrocyte-loaded β-tricalcium phosphate (β-TCP) complexes were incubated in vitro and implanted into the defects of the articular cartilage in the humeral head of the goat. The specimens were taken within 3 and 6 months after surgical procedure for histological and im-munohistochemical evaluation. Results In experimental group, defects were filled with cartiaginous tissue entirely. Histological and immunohistochemical analysis showed extensively cartilage formation in the defect field and the staining for II collagen was positive obviously. In the β-TCP group, the defects were partly repaired by new-formed cartilage around the defects and there is no cartilage formation in the scaffold center. In the control groups,only a little new-formed cartilage can be demonstrated around the margin of defects.There is a significant difference between the experimental and the other two groups(P< 0.01) .Conclusions Immortalized chondrocytes may have a significant application in the cartilage tissue-engineering field.