胶原杂化去抗原松质骨载体复合自体间充质干细胞修复骨软骨缺损

Collagen hybridized antigen-extracted cancellous bone scaffold supplemented with autologous mesenchymal stem cells to treat osteochondral defect

目的制备胶原杂化去抗原松质骨载体并复合自体间充质干细胞修复关节骨软骨缺损。方法脱脂、脱蛋白和脱钙的方法来制备去抗原松质骨载体,酸解-酶解法制备胶原进而杂化松质骨载体。扩增间充质干细胞,接种到胶原杂化松质骨载体,植入左膝关节(实验侧),右膝关节植入没有复合细胞的胶原杂化去抗原松质骨载体(对照侧),在第6、12、24、48周时结合Pineda评分观察实验侧和对照侧的骨软骨缺损的组织修复情况。结果实验侧自12周时出现初步的软骨表型,24周时最为明显,48周有部分退变;软骨下骨部分逐渐恢复。对照侧软骨缺损一直以纤维组织充填,软骨下骨有部分恢复。上述各时间点实验侧和对照侧的Pineda评分为16.600±0.966,17.600±0.699(6周);6.600±0.M3、14.800±0.789(12周);3.300±0.483、13.600±0.843(24周);3.500±0.527、14.100±0.876(48周),差异均有统计学意义(P<0.05)。结论胶原杂化的去抗原松质骨载体复合自体间充质干细胞能够修复关节的骨软骨缺损。

Objective To prepare collagen hybridized antigen-extracted cancellous bone supplemented with autologous mesenchymal stem cells and apply it to treat osteochondral defect. Methods Antigen-extracted cancellous bone was prepared by degreasing, deproteinization and decalcification. Collagen type I was produced by means of acidolysis and enzymolysis and used to hybridize the resulting cancellous bone scaffold. Antologous mesenchymal stem cells were harvested, expanded and seeded in the collagen hybridized antigen-extracted cancellous bone scaffold to repair osteochondral defect in the left knee ; and as an auto-control,the right knee of the same rabbit was implanted with a same scaffold that was cell-free. Reparative profile of the defect was assessed by histological examination and Pineda score at regular intervals of 6th, 12th,24th and 48th week. Results The reparative tissue on the experimental side began to take on chondrogenic phenotype at 12th week,improved at 24th week and degraded partially at 48th week, and the subchondral bone was restored gradually. However, on the control side,the defects were filled with fibrous tissue throughout the 48 week and the subchondral bone was not reconstructed completely. Pineda score revealed statistical difference between the experimental and the control sides at each time point, namely, 16. 600 ± 0.966 va 17. 600 ± 0. 699 （6th week） ; 6. 600 ± 0. 843 vs 14. 800 ± 0. 789 （ 12th week） ; 3. 300 ±0.483 vs 13. 600 ±0.843 （24th week） ;3. 500 ± 0. 527 vs 14. 100 ± 0. 876 （48th week） respectively. Conelusion The collagen hybridized antigen-extracted cancellous bone scaffold supplemented with autologous mesenchymal stem ceils can be used to treat osteochondral defect without osteogenic and chondrogenic induction in vitro.