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首页> 外文期刊>BioResearch open access. >Construction of Mesenchymal Stem Cell–Containing Collagen Gel with a Macrochanneled Polycaprolactone Scaffold and the Flow Perfusion Culturing for Bone Tissue Engineering
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Construction of Mesenchymal Stem Cell–Containing Collagen Gel with a Macrochanneled Polycaprolactone Scaffold and the Flow Perfusion Culturing for Bone Tissue Engineering

机译：大通道聚己内酯支架的间充质干细胞胶原凝胶的构建及骨组织工程的血流灌注培养

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A novel bone tissue-engineering construct was developed by using poly(?-caprolactone) (PCL)-macrochanneled scaffolds combined with stem cell-seeded collagen hydrogels and then applying flow perfusion culture. Rat mesenchymal stem cells (MSCs) were loaded into collagen hydrogels, which were then combined with macrochanneled PCL scaffolds. Collagen hydrogels were demonstrated to provide favorable growth environments for MSCs and to foster proliferation. Cell number determination identified retention of substantially fewer (50–60%) cells when they were seeded directly onto macrochanneled PCL than of cells engineered within collagen hydrogels. Additionally, the cells actively proliferated within the combined scaffold for up to 7 days. MSC-loaded collagen–PCL scaffolds were subsequently cultured under flow perfusion to promote proliferation and osteogenic differentiation. Cells proliferated to levels significantly higher in flow perfusion culture than that under static conditions during 21 days. A quantitative polymerase chain reaction (QPCR) assay revealed significant alterations in the transcription of bone-related genes such as osteopontin ( OPN ), osteocalcin ( OCN ), and bone sialoprotein ( BSP ), such as 8-, 2.5-, and 3-fold induction, respectively, after 10 days of flow perfusion relative to those in static culture. OPN and OCN protein levels, as determined by Western blot, increased under flow perfusion. Cellular mineralization was significantly enhanced by the flow perfusion during 21 and 28 days. Analyses of mechanosensitive gene expression induced by flow perfusion shear stress revealed significant upregulation of c-fos and cyclooxygenase-2 (COX-2) during the initial culture period (3–5 days), suggesting that osteogenic stimulation was possible as a result of mechanical force-driven transduction. These results provide valuable information for the design of a new bone tissue-engineering system by combining stem cell-loaded collagen hydrogels with macrochanneled scaffolds in flow perfusion culture.
机译：通过使用聚（ε-己内酯）（PCL）的微通道支架与干细胞接种的胶原水凝胶结合，然后应用流灌注培养，开发了一种新型的骨组织工程构建体。将大鼠间充质干细胞（MSC）加载到胶原蛋白水凝胶中，然后与大通道PCL支架结合。胶原蛋白水凝胶被证明可为MSC提供有利的生长环境并促进增殖。确定细胞数量后，直接植入大通道PCL的细胞比保留在胶原水凝胶中的细胞要少得多（50-60％）。另外，细胞在组合支架内活跃增殖长达7天。随后在流动灌注下培养装载MSC的胶原蛋白PCL支架，以促进增殖和成骨分化。在21天的流动灌注培养中，细胞的增殖水平明显高于静态条件下的水平。定量聚合酶链反应（QPCR）分析显示，与骨相关的基因（如骨桥蛋白（OPN），骨钙蛋白（OCN）和骨唾液蛋白（BSP），如8、2.5-和3-的转录明显改变相对于静态培养中的血流灌注，经过10天的血流灌注后，它们的诱导倍数分别为。通过蛋白质印迹确定的OPN和OCN蛋白水平在血流灌注下增加。在21和28天内，血流灌注显着增强了细胞矿化作用。血流灌注剪切应力诱导的对机械敏感基因表达的分析显示，在初始培养期间（3-5天），c-fos和环氧合酶-2（COX-2）明显上调，这表明机械刺激可导致成骨性刺激力驱动的转导。这些结果通过在流灌注培养中将干细胞负载的胶原蛋白水凝胶与大通道支架相结合，为设计新的骨组织工程系统提供了有价值的信息。

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