用旋转生物反应器体外再造骨组织的实验研究

Experimental Research of Tissue Engineered Bone Formation in Vitro in a Spinner Flask Bioreactor System

目的:将骨髓基质干细胞接种于聚乙醇酸支架中,在旋转生物反应器内进行动态培养,观察骨组织的形成情况。方法:分离、培养、扩增猪骨髓基质干细胞,细胞长满后用地塞米松诱导,收集细胞,按5×107/mL的浓度50μL接种于聚乙醇酸支架中,静态培养5 d后,移入旋转生物反应器内进行动态培养,2月取材,通过大体标本、组织学检查观察新骨的形成情况,以静态培养作为对照。结果:细胞接种后1 d,即可以贴附于材料表面,显微镜下观察,细胞在材料表面伸展良好。连续培养2个月,形成质地坚硬的组织;组织学检查可见新形成的骨样组织,厚度约100μm,而对照组中在材料表面有纤维组织形成。结论:旋转生物反应器内进行动态培养是体外再造骨组织的有效手段。

Objective： To evaluate in vitro new bone formation by bone marrow mesenchymal stem cells （MSCs） seeded into polyglycolic acid （PGA） scaffold in a spinner flask bioreactor system. Methods： Bone marrow mesenchymal stem cells （MSCs） were cultured, expanded and induced in vitro. Cells were seeded into PGA non -woven mesh at the density of 5 × 10^7/ml in 50μl cell suspension. Cell growth on the scaffold was observed by phase - contrast microscope. After 5 days static incubation, four cell -polymer complexes were transferred into a spinner flask bioreactor. Two months later, bone formation was observed by gross inspection examination and histological observation. Four cell -polymer complexes were incubated in static condition acting as control. Results： MSCs adhered and spreaded well on the surface of PGA scaffold. Hard tissue was restored successfully in the bioreactor 2 months after implantation. And histological examination demonstrated that newly restored tissue was composed of osteoid. In the control group only soft tissue formed on the surface of the scaffold. Conclusion： It was an efficient method to form new bone in vitro by seeded MSCs into PGA scaffold and incubated in a spinner flask bioreactor system.