Institute of Biomedical Engineering, College of Medicine and College of Engineering, National Taiwan University, Taipei 100, Taiwan.
J Biomed Mater Res A. 2009 Dec;91(3):929-38. doi: 10.1002/jbm.a.32309.
The aim of this study was to evaluate the behaviors of mesenchymal stem cells (MSCs) on Poly(L-Lactic acid) (PLLA) membranes with different surface topographies. The double-sided micropatterns, island-patterned, and sunken-patterned PLLA membranes with diameters of 60 and 100 microm, were fabricated by the soft lithography method. The cell viability of MSCs on the island- and sunken-patterned PLLA membranes were characterized by scanning electron microscopy (SEM), MTT assay, and flow cytometric analysis. Cell adhesion and proliferation capability were superior for the MSCs seeding on the island-patterned PLLA membranes than those on the sunken-patterned PLLA membranes. Especially, we observed the best biocompatibility for MSCs on the island-patterned surface with diameter of 100 microm. In addition, the improvement of cell attachment and augmenting subsequent cellular response are investigated after the island-patterned membranes precoating with collagen and fibronectin. Furthermore, the flow cytometric analysis reveals the MSCs can expand and maintain the phenotype on these PLLA membranes without losing its potential for differentiation. Since scale-up of cell production and optimization of culture conditions are important for stem cell engineering, to control the stem cell proliferation and differentiation is necessary. Therefore, besides topographical properties play a crucial role on the stem cells attachment and proliferative activity, it is suggested that the "relative scale" between cell and pattern also affects the cell adhesion morphologies and cell behaviors. Based on the overall cellular response, this study provides a valuable guidance to prepare appropriate topographic surface for tissue engineering application.
本研究旨在评估间充质干细胞（MSCs）在具有不同表面形貌的聚（L-乳酸）（PLLA）膜上的行为。通过软光刻法制备了具有 60 和 100 微米直径的双面微图案、岛图案和凹坑图案 PLLA 膜。通过扫描电子显微镜（SEM）、MTT 测定和流式细胞术分析来表征 MSC 在岛状和凹坑状 PLLA 膜上的细胞活力。与凹坑状 PLLA 膜相比，MSC 在岛状 PLLA 膜上的黏附和增殖能力更强。特别是，我们观察到 100 微米直径的岛状表面具有最佳的生物相容性。此外，在岛状膜上预先涂覆胶原蛋白和纤连蛋白后，研究了细胞附着的改善和随后的细胞反应增强。此外，流式细胞术分析表明，MSC 可以在这些 PLLA 膜上扩增并保持其表型，而不会失去其分化潜能。由于细胞生产的规模化和培养条件的优化对于干细胞工程非常重要，因此控制干细胞的增殖和分化是必要的。因此，除了形貌特性对干细胞的附着和增殖活性起着至关重要的作用外，还建议细胞与图案之间的“相对比例”也会影响细胞的黏附形态和细胞行为。基于整体细胞反应，本研究为组织工程应用制备合适的形貌表面提供了有价值的指导。
