Department of Biomedical Engineering, Tufts University, Medford, Massachusetts 02155, USA.
J Biomed Mater Res A. 2011 Jan;96(1):21-8. doi: 10.1002/jbm.a.32949. Epub 2010 Oct 14.
Tissue engineered cartilage can be grown in vitro with the use of cell-scaffold constructs and bioreactors. The present study was designed to investigate the effects of perfusion bioreactors on the chondrogenic potential of engineered constructs prepared from porous silk fibroin scaffolds seeded with human embryonic stem cell (hESC)-derived mesencyhmal stem cells (MSCs). After four weeks of incubation, constructs cultured in perfusion bioreactors showed significantly higher amounts of glycosaminoglycans (GAGs) (p < 0.001), DNA (p < 0.001), total collagen (p < 0.01), and collagen II (p < 0.01) in comparison to static culture. Mechanical stiffness of constructs increased 3.7-fold under dynamic culture conditions and RT-PCR results concluded that cells cultured in perfusion bioreactors highly expressed (p < 0.001) cartilage-related genes when compared with static culture. Distinct differences were noted in tissue morphology, including polygonal extracellular matrix structure of engineered constructs in thin superficial zones and an inner zone under static and dynamic conditions, respectively. The results suggest that the utility of perfusion bioreactors to modulate the growth of tissue-engineered cartilage and enhance tissue growth in vitro.
组织工程软骨可以通过细胞-支架构建体和生物反应器在体外生长。本研究旨在研究灌注生物反应器对多孔丝素支架接种人胚胎干细胞（hESC）衍生间充质干细胞（MSCs）构建的工程化构建体的软骨形成潜力的影响。孵育四周后，与静态培养相比，灌注生物反应器中培养的构建体的糖胺聚糖（GAG）（p < 0.001）、DNA（p < 0.001）、总胶原（p < 0.01）和胶原 II（p < 0.01）含量显著增加。在动态培养条件下，构建体的机械刚度增加了 3.7 倍，并且 RT-PCR 结果表明，与静态培养相比，在灌注生物反应器中培养的细胞高度表达（p < 0.001）与软骨相关的基因。组织形态学上存在明显差异，包括工程化构建体的薄表层中的多边形细胞外基质结构和静态和动态条件下的内层。结果表明，灌注生物反应器可用于调节组织工程软骨的生长并增强体外组织生长。
