自体骨髓基质干细胞及可注射藻酸钙凝胶联合Mosaicplasty技术修复山羊膝关节股骨头大面积缺损

Repairing large area osteochondral defects using Mosaicplasty modified by autologous bone mesenchymal stem cells combined with alginate calcium gel

背景：目前修复软骨缺损的方法都存在修复组织数量不足，生物力学性能不佳，整合不良及供区并发症等缺陷。对于大面积的骨软骨复合缺损单独应用一种方法尚显不足。目的：观察组织工程方法复合Mosaicplasty技术用于修复大面积骨软骨缺损的效果。设计、时间及地点：随机对照动物实验，于2009—0I／09在青岛大学医学院中心实验室完成。材料：体外培养并扩增健康成年雄性山羊的骨髓基质干细胞，收集约3×10^7细胞，加入1mL藻酸钠溶液重恩形成骨髓丛质干细胞-藻酸钙凝胶材料。方法：12只山羊用于制备膝关节股骨髁大面积骨软骨缺损模型，骨髓基质干细胞Mosaicplasty组使用自制Mosaicplasty器械，植入直径2mm骨软骨柱镶嵌充填缺损，以自体骨髓基质干细胞复合藻酸钙凝胶填充残余缺损和部分供区。Mosaicplasty组单纯用Mosaicplasty修复骨软骨缺损。对照组单纯制造缺损不修复。主要观察指标：①大体观察：术后4，8，16周分别切开关节观察修复效果。②组织学检查：术后16周取修复组织标本，行苏木精-伊红染色、甲苯胺蓝染色光镜下观察。③电镜观察：取16局修复组织行透射电镜检查。结果：术后16周时骨髓基质干细胞-Mosaicplasty组移植物固定牢固，关节面平滑，移植物间界限消失，新生软骨组织类似于正常软骨，4-16周修复效果逐渐改善，优于其他各组。光镜观察细胞-凝胶新生软骨组织与移植软骨结合紧密，新生软骨细胞排列规整，细胞外基质分布均。对照组无明显修复。透剔电镜观察发现修复新生组织中细胞形态类似软骨细胞，细胞存在于排列紧密的纤维网格中，基质丰富。结论：使用自体骨髓基质干细胞-藻酸钙凝胶材料复合Mosaicplasty技术可促进骨软骨整合．改善其修复效果。

BACKGROUND： The existed repair method for cartilage defects has shortcomings of insufficient repairing tissue numbers, poor biomechanical properties, as well as donor site complication. Thus it is deficient to repair large-sized osteochondral defects using one method. OBJECTIVE： To investigate the therapeutic effect of tissue engineering modified Mosaicplasty on repairing large-sized osteochondral defects. DESIGN, TIME AND SETTING： A randomized controlled animal experiment was performed at the Center Laboratory of Qingdao University Medical College from January to September 2009. MATERIALS： The hircine bone mesenchymal stem cells （BMSCs） were in vitro cultured, and resuspended with algin solution to obtain BMSCs-calcium alginate gel. METHODS： Totally 12 goats were prepared for osteochondral defects models and were divided into 3 groups. BMSCs-Mosaicplasty group, BMSCs compound with injectable alginate calcium gel was then applied to fill the ＂dead space＂ after Mosaicplasty. In the Mosaicplasty group, the defects were repaired by Mosaicplasty. There was no treatment in the control group. From 4 to 16 weeks postoperatively, the animals were sacrificed and the in gross and under electromicroscopy. MAIN OUTCOME MEASURES： （1）Gross observation： the joint was exposed to observe the repair effect at weeks 4, 8, 16 after operation. （2）Histological examination： specimens were harvested at 16 weeks after operation and observed by haematoxylin-eosin staining, toluidine blue staining under light microscopy. （3）Transmission electron microscope was used at 16 weeks after operation. RESULTS： The transplanted subchondral bone and superficial cartilage was integrated hardly with each other or with recipient sites in tissue engineering modified Mosaicplasty groups at 16 weeks after operation. The quality and appearance of the transplanted and regenerated cartilage was similar to normal hyaline cartilage. Under microscopy, the regenerated cartilage was integrated with neighbor tightly in regular arrange. ECM distributed evenly and deeply stained by alcian blue. There was no obviously repaired in the control group. CONCLUSION： Tissue engineering can ameliorate the outcome of Mosaicplasty to repair the osteochondral defects.