Tubular perfusion system for chondrocyte culture and superficial zone protein expression.
Tubular perfusion system for chondrocyte culture and superficial zone protein expression.
复制标题
用于软骨细胞培养和浅表区蛋白表达的管状灌注系统。
DOI：
10.1002/jbm.a.35321
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发表时间：
2015
期刊：
影响因子：
--
通讯作者：
Fisher,JohnP
中科院分区：
文献类型：
--
作者：
Yu,Li;Ferlin,KimberlyM;Nguyen,Bao-NgocB;Fisher,JohnP
组织工程是关节软骨修复的另一种方法。机械刺激已被发现是软骨细胞健康发育和维持其原生表型的重要因素。为了增强营养转运和施加机械应力，我们开发了一种新的生物反应器，管状灌注系统（TPS），用于在三维支架中培养软骨细胞。在我们的设计中，软骨细胞被包裹在藻酸盐支架中，并放置在管状生长室中，该生长室中灌注介质以增强营养物质的转移并使细胞暴露于流体中。结果显示，DNA含量和组织化学染色显示，与静态培养相比，TPS培养促进软骨细胞增殖。培养14天后，TPS生物反应器培养中促炎和凋亡标志物的低信使RNA表达证实，流速为3 mL/min不会损伤海藻酸盐支架内的软骨细胞。此外，与静态组相比，在TPS生物反应器中培养的细胞显示聚集蛋白、II型胶原和浅区蛋白的基因表达水平增加，表明浅区特异性表型的出现。因此，TPS生物反应器是体外增强软骨细胞增殖和表型维持的有效手段。©2014 Wiley期刊公司[J] .中国生物医学工程学报，2015,31(1):464 - 467。
Tissue engineering is an alternative method for articular cartilage repair. Mechanical stimulus has been found to be an important element to the healthy development of chondrocytes and maintenance of their native phenotype. To enhance nutrient transport and apply mechanical stress, we have developed a novel bioreactor, the tubular perfusion system (TPS), to culture chondrocytes in three‐dimensional scaffolds. In our design, chondrocytes are encapsulated in alginate scaffolds and placed into a tubular growth chamber, which is perfused with media to enhance nutrient transfer and expose cells to fluid flow. Results demonstrate that TPS culture promotes the proliferation of chondrocytes compared to static culture as shown by DNA content and histochemical staining. After 14 days of culture, low messenger RNA expression of proinflammatory and apoptotic markers in TPS bioreactor culture confirmed that a flow rate of 3 mL/min does not damage the chondrocytes embedded in alginate scaffolds. Additionally, cells cultured in the TPS bioreactor showed increased gene expression levels of aggrecan, type II collagen, and superficial zone protein compared to the static group, indicative of the emergence of the superficial zone specific phenotype. Therefore, the TPS bioreactor is an effective means to enhance the proliferation and phenotype maintenance of chondrocytesin vitro. © 2014 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 103A: 1864–1874, 2015.