Department of Clinical Biochemistry, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
Department of Tissue Engineering and Applied Cell Sciences, School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
Differentiation. 2020 Sep-Oct;115:22-29. doi: 10.1016/j.diff.2020.06.003. Epub 2020 Jul 17.
Cell morphological changes induced by micro-grooved topography have been shown to be an important regulator of smooth muscle (SM) differentiation of mesenchymal stem cells (MSCs). In addition to the micro-grooved topography, transforming growth factor-β (TGF-β) can also modulate MSCs differentiation towards smooth muscle cells (SMCs) through alterations in cell morphological characteristics. Thus, it can be hypothesized that substrate topography and TGF-β may interact to facilitate differentiation of MSCs into SMCs. In this study, we investigated the time-course cooperative effects of substrate topography and TGF-β in the regulation of SM differentiation of human MSCs. Western blotting, followed by image analysis, was performed to assess the protein expression of α-actin, h1-calponin and gelsolin. Three-way analysis of variance was employed to investigate the main effect of each independent variable, i.e. TGF-β conditioning, substrate topography and culture time, along with the interactions of these variables. Each of TGF-β, substrate topography and culture time significantly affected the protein expression of α-actin, h1-calponin and gelsolin. Overall, TGF-β conditioning of the cells and culturing the cells on the micro-grooved substrate resulted in greater protein expression of α-actin and h1-calponin, and lesser protein expression of gelsolin. In addition to the isolated effects of the variables, treatment type interacted with substrate topography and culture time to regulate the expression of the above-mentioned proteins. This study indicated the feasibility of promoting SM differentiation of human MSCs by simultaneous recruitment of micro-grooved topography and TGF-β. The findings could be of assistance when effective utilization of chemo-physical cues is needed to achieve functional SMC-like MSCs in vitro.
微槽形貌诱导的细胞形态变化已被证明是调节间充质干细胞（MSCs）向平滑肌（SM）分化的重要调控因子。除了微槽形貌外，转化生长因子-β（TGF-β）还可以通过改变细胞形态特征来调节 MSCs 向平滑肌细胞（SMCs）的分化。因此，可以假设基底形貌和 TGF-β可能相互作用，促进 MSCs 向 SMC 分化。在这项研究中，我们研究了基底形貌和 TGF-β在调节人 MSCs 向 SM 分化中的时间协同作用。通过 Western blot 分析和图像分析来评估α-肌动蛋白、h1-钙调蛋白和凝胶蛋白的蛋白表达。采用三因素方差分析来研究每个独立变量（即 TGF-β处理、基底形貌和培养时间）的主要效应以及这些变量的相互作用。TGF-β、基底形貌和培养时间都显著影响α-肌动蛋白、h1-钙调蛋白和凝胶蛋白的蛋白表达。总的来说，细胞的 TGF-β处理和在微槽基底上培养会导致α-肌动蛋白和 h1-钙调蛋白的蛋白表达增加，而凝胶蛋白的蛋白表达减少。除了变量的单独效应外，处理类型与基底形貌和培养时间相互作用，调节上述蛋白的表达。这项研究表明，通过同时募集微槽形貌和 TGF-β，促进人 MSCs 向 SM 分化是可行的。当需要有效利用化学物理线索在体外获得功能性类似 SMC 的 MSCs 时，这些发现可能会有所帮助。
