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首页> 外文期刊>Journal of biomedical materials research. Part B, Applied biomaterials. >Topography enhances Runx2 expression in outgrowing cells from iPS cell-derived embryoid bodies
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Topography enhances Runx2 expression in outgrowing cells from iPS cell-derived embryoid bodies

机译：地形增强了来自IPS细胞衍生的胚胎体的超越细胞中的Runx2表达

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摘要

The effect of differing polystyrene substrate topographies on the osteogenic potential of the outgrowing cells (OGCs) formed from mouse-induced pluripotent stem (iPS) cells (miPSCs)-derived embryoid bodies (EBs) was investigated. Polystyrene substrates were sandblasted with 25, 50, and 150 mu m aluminum oxide particles to obtain topographies with average Sa values of 0.6, 1.1, and 1.8 mu m, respectively. 3D-SEM was used to evaluate substrate's topographies. Examination was done by scanning electron microscopy (SEM), by immunocytofluorescence (ICF) analysis for vinculin, Runx2 and collagen type I, and by quantitative RT-PCR (qRT-PCR) analysis for Runx2 and collagen type I. SEM and ICF analyses revealed that surface roughness caused cells elongation (2, 6, 8, 10 times for the NT, 0.6 mu m, 1.1 mu m, and 1.8 mu m, respectively). Vinculin staining demonstrated how the Sa value affected cellular attachment to the substrate. FA points were randomly distributed on flat surfaces, but rough surfaces resulted in more concentrated FA points on the podia of the cells (11.7, 25.2, 26.7, 16.6 vinculin spots per 20 mu m(2) for the NT, 0.6 mu m, 1.1 mu m, and 1.8 mu m, respectively). qRT-PCR revealed that Runx2 expression was highest on day 16 on surfaces with Sa of 0.6 mu m and 1.1 mu m. Collagen type I expression increased from day 0 to day 16, no significance was found among the groups. In conclusion, surface topography affects cell shape and expression of early osteogenic potential in OGC, particularly surfaces with Sa values of 0.6 mu m and 1.1 mu m which showed the highest concentration of FA points on podia. These findings could be utilized in the development of inner surface topographies of scaffolds used with iPSCs. (c) 2019 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater 107B: 2288-2296, 2019.
机译：研究了不同聚苯乙烯衬底地形对由小鼠诱导的多能茎（IPS）细胞（MIPSC）形成的胚胎体（EBS）形成的外苯乙烯底物地形上的效果。聚苯乙烯底物用25,50和150μm氧化铝颗粒砂粘连，以分别获得平均SA值为0.6,1.1和1.8μm的拓扑。 3D-SEM用于评估基板的拓扑。通过扫描电子显微镜（SEM），通过vinculin，runx2和胶原蛋白I型的免疫荧光（ICF）分析来进行检查，以及Runx2和胶原型I的定量RT-PCR（QRT-PCR）分析。SEM和ICF分析显示表面粗糙度引起的细胞伸长（2,6,8,10次，分别为0.6μm，1.1μm和1.8μm）。 vinculin染色证明了SA值如何将蜂窝连接到基材上。将FA点随机分布在平坦表面上，但粗糙的表面导致细胞孔的孔径上更浓缩（11.7,25.2,26.7,16.6 vinculin每20μm（2），0.6μm，1.1 mu m，分别为1.8 mu m）。 QRT-PCR显示，RUNX2表达在第16天的表面上最高，SA为0.6μm和1.1μm。胶原蛋白类型I表达从第0天增加到第16天，在组中没有发现具有重要意义。总之，表面形貌影响OGC中的细胞形状和早期成骨潜力的表达，特别是具有0.6μm和1.1μm的SA值的表面，这显示了普罗拓的最高浓度。这些发现可以用于开发与IPSCS使用的支架的内表面拓扑结构。 （c）2019年Wiley期刊，Inc。J生物保解员B部分：Appl BioMater 107B：2288-2296,2019。

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