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首页> 外文期刊>Journal of biomedical materials research. Part B, Applied biomaterials. >In vitro osteogenic differentiation of human mesenchymal stem cells photoencapsulated in PEG hydrogels
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In vitro osteogenic differentiation of human mesenchymal stem cells photoencapsulated in PEG hydrogels

机译:PEG水凝胶光包裹人间充质干细胞的体外成骨分化

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Much research has focused on the differentiation of human mesenchymal stem cells (hMSCs) in monolayer culture; however, little is known about their differentiation potential in three-dimensional culture conditions. In this research, hMSCs were encapsulated in a photocrosslinkable, injectable scaffolding system based on poly(ethylene glycol) (PEG) hydrogels. To demonstrate the ability of hMSCs to differentiate in PEG hydrogels, cell/polymer constructs were cultured in osteogenic differentiation media to elicit an osteoblastic response. First, viability of encapsulated hMSCs up to 4 weeks in culture was investigated using a membrane integrity assay. Second, gene expression of encapsulated cells was determined with reverse transcription polymerase chain reaction (RT-PCR) as a function of media composition. After 1 week in osteogenic differentiation media, encapsulated hMSCs expressed osteonectin, osteopontin, and alkaline phosphatase, which are all characteristic of osteoblasts. Finally, von Kossa staining was used to evaluate mineralization of the PEG gels. Results support the hypothesis that hMSCs photoencapsulated in PEG hydrogels and cultured in the presence of osteogenic differentiation media are able to differentiate to osteoblasts inside the gel and mineralize the matrix. These experiments demonstrate the feasibility of using a PEG-based, photocrosslinkable system to culture and deliver human mesenchymal stem cells for bone tissue regeneration and repair.
机译:许多研究集中在单层培养中人间充质干细胞(hMSCs)的分化。然而,对于它们在三维培养条件下的分化潜力知之甚少。在这项研究中,hMSCs被封装在基于聚乙二醇(PEG)水凝胶的可光交联,可注射的支架系统中。为了证明hMSC在PEG水凝胶中分化的能力,在成骨分化培养基中培养细胞/聚合物构建体以引发成骨细胞应答。首先,使用膜完整性测定法研究了封装的hMSC在培养中长达4周的生存力。其次,通过逆转录聚合酶链反应(RT-PCR)确定封装细胞的基因表达,该表达是培养基组成的函数。在成骨分化培养基中培养1周后,封装的hMSC表达了成骨细胞的特征性骨连接素,骨桥蛋白和碱性磷酸酶。最后,von Kossa染色用于评估PEG凝胶的矿化作用。结果支持以下假设:将hMSCs光囊化在PEG水凝胶中并在成骨分化培养基的存在下培养能够分化为凝胶内的成骨细胞并使基质矿化。这些实验证明了使用基于PEG的光可交联系统培养和递送人间充质干细胞用于骨组织再生和修复的可行性。

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