Yang Chung-Yao, Sung Chun-Yen, Shuai Hung-Hsun, Cheng Chao-Min, Yeh J Andrew
Institute of Nanoengineering and Microsystems, National Tsing Hua University, Hsinchu 30013, Taiwan.
Sci Technol Adv Mater. 2013 Aug 13;14(4):044406. doi: 10.1088/1468-6996/14/4/044406. eCollection 2013 Aug.
This paper describes a high-throughput method for developing physically modified chitosan membranes to probe the cellular behavior of MDCK epithelial cells and HIG-82 fibroblasts adhered onto these modified membranes. To prepare chitosan membranes with micro/nanoscaled features, we have demonstrated an easy-to-handle, facile approach that could be easily integrated with IC-based manufacturing processes with mass production potential. These physically modified chitosan membranes were observed by scanning electron microscopy to gain a better understanding of chitosan membrane surface morphology. After MDCK cells and HIG-82 fibroblasts were cultured on these modified chitosan membranes for various culture durations (i.e. 1, 2, 4, 12 and 24 h), they were investigated to decipher cellular behavior. We found that both cells preferred to adhere onto a flat surface rather than on a nanopatterned surface. However, most (> 80%) of the MDCK cells showed rounded morphology and would suspend in the cultured medium instead of adhering onto the planar surface of negatively nanopatterned chitosan membranes. This means different cell types (e.g. fibroblasts versus epithelia) showed distinct capabilities/preferences of adherence for materials of varying surface roughness. We also showed that chitosan membranes could be re-used at least nine times without significant contamination and would provide us consistency for probing cell-material interactions by permitting reuse of the same substrate. We believe these results would provide us better insight into cellular behavior, specifically, microscopic properties and characteristics of cells grown under unique, nanopatterned cell-interface conditions.
本文描述了一种用于开发物理改性壳聚糖膜的高通量方法，以探究黏附在这些改性膜上的MDCK上皮细胞和HIG - 82成纤维细胞的细胞行为。为了制备具有微/纳米尺度特征的壳聚糖膜，我们展示了一种易于操作的简便方法，该方法可轻松与具有大规模生产潜力的基于集成电路的制造工艺相结合。通过扫描电子显微镜观察这些物理改性的壳聚糖膜，以更好地了解壳聚糖膜的表面形态。在将MDCK细胞和HIG - 82成纤维细胞在这些改性壳聚糖膜上培养不同的时间（即1、2、4、12和24小时）后，对它们进行研究以解读细胞行为。我们发现两种细胞都更倾向于黏附在平坦表面而不是纳米图案化表面上。然而，大多数（> 80%）的MDCK细胞呈现圆形形态，并且会悬浮在培养基中，而不是黏附在带负电纳米图案化壳聚糖膜的平面表面上。这意味着不同的细胞类型（例如成纤维细胞与上皮细胞）对不同表面粗糙度的材料表现出不同的黏附能力/偏好。我们还表明，壳聚糖膜可以重复使用至少九次而不会有明显污染，并且通过允许重复使用相同的底物，可为我们探究细胞 - 材料相互作用提供一致性。我们相信这些结果将使我们更好地了解细胞行为，特别是在独特的纳米图案化细胞界面条件下生长的细胞的微观特性。
