旋转灌注式生物反应器系统构建及在骨组织工程中的应用

Development of Rotating Perfusion Bioreactor System And Application for Bone Tissue Engineering

构建一种旋转灌注式生物反应器系统,并设计制造具有相互连通微管道结构的大段支架。生物反应器在旋转的同时实现对大段支架的灌注,系统内外气体的实时交换通过安装在反应器两端的半透膜和可透气的蠕动泵软管实现。容器内的氧气和营养物质在得到充分混合之后,连续不断地输送到支架微管道内,使黏附在微管道内的细胞获得充足营养的同时受到一定流体剪应力的刺激,调节细胞功能的发挥。该系统克服了静态培养中存在的各种缺点,改善了培养环境,增加了培养过程的可控性,有助于促进黏附在微管道内部的细胞增殖、分化和产生大量基质。将成骨细胞/支架结构体在该生物反应器系统中培养14 d后,用扫描电镜观察细胞在大段支架微管道内的生长情况,结果表明,支架微管道内有大量细胞长入。

A rotating perfuslon bioreactor system has recently been developed in our laboratory to produce 3D dynamic culture condition, and the critical-sized scaffolds with interconnected microchennels were fabricated. Gas exchange occurs by semipermeable membrane covered on each side of bioreactor and gas-permeable peristaltic pump tube. Rotation and perfusion of culture media through large scaffolds enhance well mixing and mass transport of oxygen and nutrients in the bioreactor. Osteoblastic cells attached to microchennels are exposed to a low fluid flow-induced shear stress level. This bioreactor system overcomes several defects exited in static culture condition, improves the culture environment, facilitates osteoblast proliferation, differntiation, significant matrix production and mineralization, and the controllability of culture process is enhanced. Large scaffolds/osteoblast constructs were cultured in the bioreactor system for 14 days. Osteoblastic cells attached to microchannels of scaffolds were observed under scanning electron microscope （SEM）. The results indicated that cells grew extensively in the microchennels of large scaffolds.