Influence of Laser-Designed Microstructure Density on Interface Characteristics and on Preliminary Responses of Epithelial Cells

Abstract

Current trends in designing medical and tissue engineering systems rely on the incorporation of micro- and nano-topographies for inducing a specific cellular response within the context of an aimed application. As such, dedicated studies have recently focused on understanding the possible effects of high and low density packed topographies on the behavior of epithelial cells, especially when considering their long-term viability and functionality. We proposed to use stair-like designed topographies with three different degrees of distribution, all created in polydimethylsiloxane (PDMS) as active means to monitor cell behavior. Our model cellular system was human bronchial epithelial cells (BEAS-2B), a reference line in the quality control of mesenchymal stem cells (MSCs). PDMS microtextured substrates of 4 µm square unit topographies were created using a mold design implemented by a KrF Excimer laser. Varying the spacing between surface features and their multiscale level distribution led to irregular stairs/lines in low, medium and high densities, respectively. Profilometry, scanning electron and atomic force microscopy, contact angle and surface energy measurements were performed to evaluate the topographical and interface characteristics of the designed surfaces, while density-induced cellular effects were investigated using traditional cell-based assays. Our analysis showed that microstructure topographical distribution influences the adhesion profiles of epithelial cells. Our analysis hint that epithelial organoid formation might be initiated by the restriction of cell spreading and migration when using user-designed, controlled micro-topographies on engineered surfaces.

Keywords

• polydimethylsiloxane (PDMS) replica
• laser textured mold
• BEAS cells
• topography analysis
• cellular behavior