Biomechanics Lab, Department of Mechanics, School of Aerospace Engineering, Beijing Institute of Technology, Beijing, People's Republic of China.
J Biomed Mater Res A. 2020 Oct;108(10):2080-2089. doi: 10.1002/jbm.a.36967. Epub 2020 May 4.
The topography of extracellular matrix regulates the differentiation of mesenchymal stem cells (MSCs). In particular, the effect of spreading shape or area on cellular differentiation and viability of individual MSCs cultured in the confined adhesive regions is an interesting fundamental issue. In this study, the adhesive patterns with the circularity of 0.1 or 1 and the areas of 314; 628; 1,256; or 2,512 μm were constructed using micropatterning technology. The expression of osteogenesis marker alkaline phosphatase and the apoptosis level of individual MSCs were measured using double fluorescent staining. Results indicated that individual MSCs confined in the small area showed an apoptotic tendency, and those in the large area might enter into osteogenesis. The branched shape with small circularity increased MSC viability but reduced their pluripotency compared with the circular shape. The expression of other osteogenesis markers, such as osteocalcin and Collagen I, confirmed that large and branched pattern promoted MSC osteogenesis. In addition, the transcriptional coactivator yes-associated protein (YAP) was transferred higher in the nuclei of the large and branched cells than other micropatterned groups. This study suggested that the spreading area and shape of individual MSCs regulate their viability and osteogenesis through the YAP pathway.
细胞外基质的拓扑结构调节间充质干细胞（MSCs）的分化。特别是，在有限的黏附区域中培养的单个 MSC 的铺展形状或面积对细胞分化和存活的影响是一个有趣的基本问题。在这项研究中，使用微图案化技术构建了具有 0.1 或 1 的圆形度和 314；628；1256；或 2512 μm 的面积的黏附图案。使用双荧光染色测量个体 MSC 的成骨标志物碱性磷酸酶的表达和凋亡水平。结果表明，在小面积限制的单个 MSC 表现出凋亡趋势，而在大面积限制的 MSC 可能进入成骨状态。与圆形相比，具有小圆形度的分支形状增加了 MSC 的活力，但降低了其多能性。其他成骨标志物的表达，如骨钙素和 Collagen I，证实了大的和分支的图案促进了 MSC 的成骨作用。此外，转录共激活因子 yes 相关蛋白（YAP）在大的和分支细胞的核中转移得更高，而在其他微图案化组中则较低。本研究表明，单个 MSC 的铺展面积和形状通过 YAP 途径调节其活力和成骨作用。
