纤维蛋白凝胶和脱钙骨基质支架材料复合软骨细胞修复兔膝关节软骨缺损的实验研究

目的：探讨纤维蛋白凝胶和脱钙骨基质支架材料复合软骨细胞作为软骨组织工程支架的可行性及有效性，并为后续研究可注射性材料做基础。方法：体外分离培养软骨细胞后接种到纤维蛋白凝胶和脱钙骨基质支架材料体外培养4周，然后植入兔膝关节软骨缺损区继续培养4、8、12周后取材，分别行大体、组织学、Ⅱ型胶原免疫组织化学观察。并进行Wakitani评分，观察其体内修复关节缺损效果。结果：大体观察4周后，实验组软骨缺损区可有乳白色组织修复，12周可修复完全，并无明显凹凸感。光镜下8周可见大量软骨细胞修复，并在TB染色下见Ⅱ型胶原比4周时明显增多。12周时软骨陷窝结构形成，细胞形态排列及Ⅱ型胶原与正常软骨组织相近。结论：纤维蛋白凝胶和脱钙骨基质支架材料复合软骨细胞可以作为软骨组织工程支架材料，能够用于再生修复软骨的缺损。并为构建可注射性修复材料提供途径。

Research of repairing rabbit knee joint cartilage defect by compound material of fibrin glue and decalcified bone matrix(DBM) and chondrocytes

Objective ： To investigate the feasibility and effectivity of using compound material of fibrin glue and DBM as scaffolds for cartilage tissue engineering. Methods：Chondrocytes isolated from articular cartilage were seeded into prepared scaffolds,after incubation for 4 weeks in vitro. Chondrocytes and fibrin glue and DBM constructs were implanted in the joint cave of rabbit. The specimens were excised at the 4th, 8th, 12th week, examined grossly analyzed by haematoxylin cosine, toluidine blues staining and type Ⅱ collagen immunohistochemistry reaction. Wakitani score was counted to evaluate the repairing effect. Results：Grossly analysis showed some ivory tissue filled the caves after 4 weeks and the caves were full filled with smooth surfaee after 12 weeks. The microscope showed a good deal of chondrocytes appeared after 8 weeks and more type Ⅱ collagen than 4 weeks. Twelve weeks later,cartilage lacuna could be observed. The cells arrangement and the amount of type Ⅱ collagen both showed the same as the natural one. Conclusion：Complicated material of fibrin glue and DBM as scaffolds can be used as scaffolds for cartilage tissue engineering.