Detamore Michael S, Athanasiou Kyriacos A
Department of Bioengineering, Rice University, Houston, Texas, USA.
Tissue Eng. 2005 Jul-Aug;11(7-8):1188-97. doi: 10.1089/ten.2005.11.1188.
This objective of this study was to determine the effects of a rotating bioreactor in temporomandibular joint (TMJ) disc tissue engineering. Porcine TMJ disc cells were seeded at a density of 20 million cells/mL onto nonwoven poly(glycolic acid) (PGA) scaffolds in spinner flasks for 1 week and then cultured either under static conditions or in a rotating bioreactor for a period of 6 weeks. A series of analyses was performed, including mechanical testing, measurement of cellularity, quantification of matrix biosynthesis with a hydroxyproline assay and enzyme-linked immunosorbent assays, and observation of matrix distribution with immunohistochemistry. Between the bioreactor and static cultures, there were marked differences in gross appearance, histological structure, and distribution of collagen types I and II. Engineered constructs from the bioreactor contracted earlier and to a greater extent, resulting in a denser matrix and cell composition. In addition, immunostaining intensity was generally uniform in static constructs, in contrast to higher intensity around the periphery of bioreactor constructs. Moreover, bioreactor constructs had higher amounts of collagen II than did static constructs. However, differences in total matrix content and compressive stiffness were generally not significant. On the basis of the results of this study there is no clear benefit from use of the rotating bioreactor, although a sequence of static culture followed by rotating bioreactor culture may prove in the future to be more beneficial than either alone.
本研究的目的是确定旋转生物反应器在颞下颌关节（TMJ）盘组织工程中的作用。将猪TMJ盘细胞以2000万个细胞/毫升的密度接种到转瓶中的非织造聚乙醇酸（PGA）支架上，培养1周，然后在静态条件下或旋转生物反应器中培养6周。进行了一系列分析，包括力学测试、细胞计数、用羟脯氨酸测定法和酶联免疫吸附测定法定量基质生物合成，以及用免疫组织化学观察基质分布。生物反应器培养物和静态培养物在总体外观、组织结构以及I型和II型胶原分布方面存在明显差异。来自生物反应器的工程构建物收缩更早且程度更大，导致基质和细胞组成更致密。此外，静态构建物中的免疫染色强度通常均匀，而生物反应器构建物周边的免疫染色强度更高。而且，生物反应器构建物中的II型胶原含量高于静态构建物。然而，总基质含量和压缩刚度的差异通常不显著。基于本研究结果，使用旋转生物反应器没有明显益处，尽管未来可能证明先进行静态培养然后进行旋转生物反应器培养的序列比单独使用任何一种方法更有益。
