旋转生物反应器内微载体扩增骨髓间充质干细胞组织工程法修复兔下颌骨缺损

Rapidly proliferated bone marrow mesenchymal stem cells (BMSCs) in vitro by microcarrier vehicle in rotating bioreactor form bone for tissue engineering and repair mandible defects

目的探索旋转生物反应器内微载体扩增骨髓间充质干细胞(BMSCs)组织工程法及BM-SCs与PHB的复合支架修复兔下颌骨缺损的可行性。方法体外分离纯化BMSCs,在旋转生物反应器内,利用微载体将其在短时间内快速扩增后接种于聚羟基丁酸酯(PHB)支架上,以修复兔的下颌骨缺损区,此为实验A组;以未对缺损区进行修复为对照B组;以单纯PHB修复缺损区为对照C组。分别于修复术后第3、8、14、42、84天处死每组兔2只,对缺损区行组织学检查、骨形态发生蛋白(BMP)免疫组织化学检查、X线摄片。结果第84天,A组的大部分修复材料被骨性组织取代;B组的缺损区未修复;A组修复骨缺损的效率较C组高。结论在旋转生物反应器内,应用微载体技术可以成功地进行BMSCs的培养和快速扩增,能满足骨组织工程的需要;PHB可以作为一种组织工程材料修复骨缺损。

Objective To investigate the rapid proliferation method of rabbit bone marrow mesenchyrnal stern cells （BMSCs） cultured by mierocarrier in a rotating bioreactor. Methods BMSCs of rabbit were separated and purifled in vitro, and rapidly proliferated by mierocarrier in a rotating bioreactor in a short period of time. The BMSCs were seeded into polyhyclroxybutyrate （PHB）, and then implanted into the mandible defect, to evaluate the performance outcome of experimental group A versus control group B with simple mandible defect, and compare group A with control group C （PHB alone）. Two rabbits from each group were killed on days 3, 8, 14, 42, 84, and specimeas were evaluated by histological examination, BMP immunohistochemistry and X - ray. Results Most PHB were replaced by bone tissue in the experimental group at day 84 after implantation, and the reconstruction of defects was superior to the control group C, but there was only fibrous tissue in control group B. Conclusion BMSCs can be cultured and rapidly proliferated by mierocarrier vehicle in a rotating bioreactor, and this technique can meet the needs of bone tissue engineering. PHB can be one of use as a repair material for defects requiring bone tissue engineering.