双腔搅拌式生物反应器提高β-磷酸三钙修复关节软骨缺损的效果

Double-chamber stirred bioreactor improves the repaired effect of beta-tricalcium phosphate on goat knee cartilage defects

背景:前期实验自行研发了一种可进行成骨及成软骨双向诱导分化的双腔搅拌式生物反应器。目的:探索双腔搅拌式生物反应器的力学刺激能否提高组织工程骨软骨修复山羊膝关节缺损的效果。方法:取青山羊12只,制作双侧后肢股骨内髁骨软骨缺损,随机分组,实验组与对照组缺损处均植入在双腔搅拌式生物反应器中进行成软骨、成骨诱导2周的自体骨髓间充质干细胞与β-磷酸三钙复合体,不同的是实验组将双腔搅拌式生物反应器置于磁力搅拌仪上给予力学刺激,对照组未给予力学刺激;空白对照组不做处置。植入后12,24周进行大体观察,Masson染色、Ⅱ型胶原免疫组织化学染色和组织学评分。结果与结论:实验组与对照组均有新生软骨与骨组织生成,实验组修复骨缺损效果明显优于对照组(P<0.05);空白对照组无新生软骨生成。表明通过双腔搅拌式生物反应器体外培养阶段的力学刺激,可以改善以山羊骨髓间充质干细胞为种子细胞、β-磷酸三钙为支架的组织工程骨软骨复合物对膝关节缺损的修复效果。

BACKGROUND： A preliminary experiment developed a double-chamber stirred bioreactor which can carry out osteogenic and cartilage induction at the same time. OBJECTIVE： To explore the effects of double-chamber stirred bioreactor on the repair of goat knee cartilage defects with tissue-engineered cartilage. METHODS： Twelve goats were selected to make bilateral femoral condyle osteochondral defects models and randomized to three groups： experimental group, implanted with the composites of β-tricalcium phosphate and bone marrow mesenchymal stem cells that were subjected to 2-week chondrogenic and osteogenic induction simultaneously in the double-chamber stirred bioreactor under mechanical stimulation; control group, implanted with the composites of β-tricalcium phosphate and bone marrow mesenchymal stem cells that were subjected to 2-week chondrogenic and osteogenic induction simultaneously in the double-chamber stirred bioreactor; blank control group, without treatment. After 12 and 24 weeks of implantation, general observation, Masson staining, II collagen immunohistochemical staining and histological scoring were performed. RESULTS AND CONCLUSION： In the experimental and control groups, new cartilage tissue and bone tissue were visible, but the experimental group showed better repair effects than the control group （P 〈 0.05）. The blank control group had no cartilage formation. These findings indicate that under the mechanical stimulation by the double-chamber stirred bioreactor in vitro, the repair effect of tissue-engineered osteochondral complex on knee joint cartilage defects can be improved.