Department of Tissue Engineering and Applied Cell Sciences, School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
Stem Cell Technology Research Center, Tehran, 1997775555, Iran.
J Cell Biochem. 2018 Jan;119(1):625-633. doi: 10.1002/jcb.26223. Epub 2017 Jul 17.
Scaffolds porosity has an important role in in vitro and in vivo differentiation process of stem cells with given the amount of space available to the cells to proliferate and differentiate. In the present study, chitosan with three porosities including 10%, 15%, and 20% that created by gelatin were used for investigation of the proliferation and osteogenic differentiation potential of adipose-derived stem cells (ADSCs). In order to be more like the scaffold to natural bone tissue, freeze-drying method was used in the scaffold preparation. Scaffold morphology, cell attachment, and toxicity were evaluated using scanning electron microscopy and MTT assay. Then, osteogenic differentiation potential of ADSCs cultured on chitosan with different porosities was evaluated by common osteogenic markers such as Alizarin red staining, ALP activity, calcium content, and osteogenic-related genes expression via real-time RT-PCR. Although all scaffolds supported the proliferation and differentiation of ADSCs, but 10% scaffold demonstrated higher amount of osteogenic markers in comparison with the other porosities and control groups. Taking together, it can be concluded that osteogenic differentiation well done in the scaffolds with lower porosity because density of the cells will increase by forcing resulted from the scaffold, so osteogenic differentiation of the stem cells have an inverse association with scaffold porosity. J. Cell. Biochem. 119: 625-633, 2018. © 2017 Wiley Periodicals, Inc.
支架的孔隙率在干细胞的体外和体内分化过程中起着重要作用，因为它为细胞提供了增殖和分化所需的空间。在本研究中，使用了三种不同孔隙率的壳聚糖（10%、15%和 20%），这些壳聚糖是通过明胶形成的，用于研究脂肪来源干细胞（ADSCs）的增殖和成骨分化潜力。为了使支架更接近天然骨组织，采用冷冻干燥法制备支架。通过扫描电子显微镜和 MTT 测定评估支架形态、细胞附着和毒性。然后，通过茜素红染色、碱性磷酸酶（ALP）活性、钙含量和实时 RT-PCR 检测成骨相关基因表达等常用成骨标志物评估不同孔隙率壳聚糖上 ADSCs 的成骨分化潜力。虽然所有支架都支持 ADSCs 的增殖和分化，但与其他孔隙率和对照组相比，10%的支架表现出更高的成骨标志物含量。综上所述，成骨分化在孔隙率较低的支架中完成得更好，因为细胞密度会因支架的强制作用而增加，因此干细胞的成骨分化与支架孔隙率呈负相关。J. Cell. Biochem. 119: 625-633, 2018. © 2017 Wiley Periodicals, Inc.
