Sun Jun, Hou Xiao-kui, Kuang Yong, Wei Xiao-en, Shi Meng
Department of Orthopaedics, Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai 200021, China.
Zhongguo Gu Shang. 2011 Sep;24(9):768-74.
To investigate the optimum proportion of Mosaicplasty and genes-enhanced tissue engineering for the repair of acute osteochondral defects.
Western blot test was conducted to detect the expression of hTGF-beta1, Col II and Aggrecan in 3 groups, including hTGF-beta1, transduction group, Adv-betagal transduction group and control group without transduction. Eighteen 6-month-old Shanghai male goats (weight: 22 to 25 kg) were used. BMSCs were isolated from the autologous bone marrow, and were subcultured to get the cells at passage 3. Thirty-six medial femoral condyles were used and divided into 6 groups named AR, AL, BR, BL, CR, and CL. Acute cylindrical defects (9 mm in diameter and 3 mm in depth)were created in the weight bearing area of the medial femoral condyle of hind limbs. In the single group, the autologous osteochondral mosaicplasty was performed to repair the defect; in the combination group, besides the mosaicplasty, the dead space between the cylindrical grafts and the host cartilage were injected with the suspension of hTGF-beta1, gene enhanced autogenous BMSCs in sodium alginate, and CaCl2 was dropped into it to form calcium alginate gels. The autologous osteochondral transplantation cover rates of group AR was 44.44% single group, AL was 44.44% combination group, BR was 33.33% single group, BL was 33.33% combination group, CR was 22.22% single group, and CL was 22.22% combination group. The goats were killed 24 weeks after operation to receive gross and histology observation, which was evaluated by the histological grading scale of O'Driscoll, Keeley and Salter. Immunohistochemistry and TEM observation were also performed.
Western blot test showed the expression of the hTGF-beta1, Col II and the Aggrecan in the hTGF-beta1 transduction group were significantly higher than that of the Adv-betaga1 transduction and the blank control groups. The gross and histology observation revealed that each defects of six groups had different degrees of repairing. There was no significantly difference among the BL, AR, and AL groups. But the scores of the other three groups (BR, CR, and CL) were significantly poorer than the former three groups.
Mosaicplasty associated with genes enhanced tissue engineering could repair the osteochondral defects effectively. With the autologous osteochondral transplantation coverage reducing, the advantage of the combination could have a better representation.
探讨镶嵌成形术与基因增强组织工程修复急性骨软骨缺损的最佳比例。
采用蛋白质免疫印迹法检测3组（hTGF-β1转导组、Adv-betagal转导组和未转导的对照组）中hTGF-β1、Ⅱ型胶原（Col II）和聚集蛋白聚糖（Aggrecan）的表达。选用18只6月龄上海雄性山羊（体重22～25 kg）。从自体骨髓中分离出骨髓间充质干细胞（BMSCs），传代培养至第3代。取36个内侧股骨髁，分为AR、AL、BR、BL、CR、CL 6组。在山羊后肢内侧股骨髁的负重区制造急性圆柱形缺损（直径9 mm，深3 mm）。单一组采用自体骨软骨镶嵌成形术修复缺损；联合组除镶嵌成形术外，在圆柱形移植物与宿主软骨之间的死腔内注入hTGF-β1、基因增强的自体BMSCs海藻酸钠悬液，并滴加CaCl2形成海藻酸钙凝胶。AR组自体骨软骨移植覆盖率单一组为44.44%，联合组AL为44.44%，BR组单一组为33.33%，联合组BL为33.33%，CR组单一组为22.22%，联合组CL为22.22%。术后24周处死山羊，进行大体和组织学观察，采用O'Driscoll、Keeley和Salter组织学分级标准进行评估。同时进行免疫组织化学和透射电镜观察。
蛋白质免疫印迹法检测显示，hTGF-β1转导组中hTGF-β1、Col II和Aggrecan的表达明显高于Adv-betaga1转导组和空白对照组。大体和组织学观察显示，6组缺损均有不同程度的修复。BL、AR和AL组之间无显著差异。但其他三组（BR组、CR组和CL组）的评分明显低于前三组。
镶嵌成形术联合基因增强组织工程可有效修复骨软骨缺损。随着自体骨软骨移植覆盖率的降低，联合应用的优势表现得更为明显。
