Pluronic F-127负载骨髓间充质干细胞和碱性成纤维细胞生长因子修复兔膝关节软骨缺损

目的探讨应用Pluronic F-127负载骨髓间充质干细胞（MSCs）和碱性成纤维细胞生长因子（bFGF）修复兔膝关节软骨缺损的可行性。方法抽取青紫蓝兔骨髓,行MSCs原代及传代培养,将3代MSCs作为种子细胞。将32只青紫蓝兔造成双膝关节4 mm×4 mm关节软骨缺损模型,随机分为四组,实验组缺损内植入负载了MSCs和bFGF的Plu-ronic F-127;细胞组植入负载了MSCs的Pluronic F-127;生长因子组植入负载了bFGF的Pluronic F-127;对照组缺损不做处理。于术后4周和16周,处死动物取材,进行大体观察、HE染色、甲苯胺蓝染色,且对各组修复组织行组织学评分及比较。结果术后4周时实验组与其他三组比较修复效果不明显;术后16周,大体观察及HE染色示实验组缺损已不易辨认,以透明样软骨修复为主,Pluronic F-127基本降解仅残余少量颗粒,甲苯胺蓝染色示细胞周围基质有较多异染质,而细胞组和生长因子组缺损表面不平整,界限清晰,修复组织以纤维组织为主,仅周缘见少许纤维软骨出现,对照组仅缺损底部有纤维组织覆盖。根据Wakitani评分标准术后16周时实验组的修复效果明显好于其他三组。结论 Plu-ronic F-127结合MSCs和bFGF的复合物最终形成类关节软骨样组织,基本修复关节软骨缺损,此方法简便易行,有实用价值,有望成为临床治疗软骨缺损的一种新方法。

Pluronic F-127 loaded with MSCs and bFGF in repairing articular cartilage defects in rabbit knee joint

Objective To study the effect of repairing articular cartilage defects in rabbit knee with Pluronic F-127 combined with Mesenchymal Stem Cells(MSCs) and basic fibroblast growth factor(bFGF).Methods Bone marrows were harevested from rabbit,MSCs were cultured and expanded in vitro.Thirty-two rabbits were randomly divided into four groups: experimental group,cell group,growth factor group and control group.Full thickness articular cartilage defects(4 mm in diameter and 4 mm in depth) were created mechanically in the middle trochlea groove of rabbit femur.Pluronic F-127/MSCs /bfGF,Pluronic F-127/MSCs,Pluronic F-127/bfGF were implanted into the defects respectively as experimental group,cell group,growth factor group,the control group underwent no special treatment.The defects were evaluated by means of macroscopic observation,HE staining,toluidine blue staining 4 and 16 weeks after operation.The histological grade was made by Wakitani scale.Results Experimental group displayed a little effect at 4 weeks,but at 16 weeks,showed better results,compared to the other three groups.At 16 weeks,gross observation showed that the specimens harvested from experimental group demonstrated a hyaline-like cartilage formation,and integrated satisfactorily with adjacent cartilage.The Pluronic F-127 around cells degraded besides some drops.The repair tissues of experimental group presented more metachromatic substance visualized by toluidine blue staining.The defects in cell group and growth factor group were repaired mainly with fibrous tissues which were less smooth,the surrounding part of the repair tissues was engineered fibrocartilage.A little white membrane formed at the bottom of the defects in control group.The quality of the new formed cartilage and the ability on hyaline cartilage formation of the experimental group were obviously better than those of other three groups by Wakitani scale.Conclusion These results demonstrate that the repair tissues formed from Pluronic F-127/MSCs / bfGF in experimental group are nearly articular cartilage tissues and can repair the defect of articular cartilage.This simple and easy method may be used clinically in the future.