【摘要】 目的:设计并构建一套骨组织工程用三维灌注生物反应器系统。方法:①明确骨组织工程用生物反应器系统的设计原则。②灌注小室由管状玻璃主体、聚四氟乙烯螺帽以及橡胶O型圈构建组成。整个系统通过铂处理的硅胶管以及三通和四通将3个灌注小室、双向蠕动泵、培养液贮存瓶、废液贮存瓶、接种口和空气滤膜连接构成。③研究生物反应器系统操作程序。④计算细胞/支架结构体内细胞受到的流体剪切力。结果:设计并构建了一套三维灌注生物反应器系统,既可以减轻内外扩散限制,也可以将力学刺激作用于细胞。该系统区别于其他设计的主要特点是:将细胞接种和长期培养整合于一体构建工程化组织;培养基可以完全穿过支架内部;同时,其模块式设计不仅可以增加工程化组织培养的数量,而且可以根据实验目的在不同时间点灵活方便地收获结构体。结构体内细胞所受到的流体剪切力可以根据灌注速率和支架性质进行计算。结论:所设计的三维灌注生物反应器系统在骨组织工程中具有广阔的应用前景,不仅可以构建工程化骨组织用于临床修复骨缺损,而且可以用于研究流体剪切力作用对骨细胞在三维支架中行为的影响。更多还原

【Abstract】 AIM:To design and build a three-dimensional perfusion bioreactor system used in bone tissue engineering.METHODS:①The design philosophy of the bioreactor system for bone tissue engineering was specified.②Perfusion chamber was built with glass cartridge,polytetrafluoroethylene screw top,O-ring.The whole system circuit consisting of three perfusion chambers,a two-directional peristaltic pump,a medium reservoir,a waste reservoir,a seeding port and some air filters was connected with platinum-cured silicone tubing,as well as some three-way junctions and four-way junctions.③The operation procedure of the bioreactor system was developed.④The fluid shear stress within perfused cell/scaffold constructs was calculated.RESULTS:A three-dimensional bioreactor system,which could mitigate both external and internal diffusion limitations as well as supply mechanical stress to the cultured cells,was designed and constructed.The system was characterized by its function of integrating cell seeding and long-term cultivation for the development of engineered tissues,its capability to make fluid flow through the scaffolds,and its modular design promoting the growth of multiple engineered tissue and providing flexibility in harvesting the constructs at different time points,which were different from other bioreactor designs.The fluid shear stress within perfused constructs could be calculated based on the perfusion flow rate and the characteristics of the scaffolds.CONCLUSION:The three-dimensional bioreactor system has promising applications in bone tissue engineering.It could be used to either construct engineered bone tissues for clinical repair of bone defects or explore the influences of fluid shear stress on the behavioral responses of osteocyte within three-dimensional scaffolds.更多还原