Quantitative analysis of hepatic cell morphology and migration in response to nanoporous and microgrooved surface structures

Abstract

Material surface topography is an important factor for regulating cellular behaviour. Understanding the mechanism of how surface topography influences mammalian cells is critical for the development of medical implants and tissue engineering. In this study, we investigated the influences of nanoporous and microgrooved substrates on the morphology and migration of hepatic cell line, BEL-7402 cells. Cells were cultured on nanoporous (140nm in diameter) anodized alumina membrane (AAM), nanoporous (140nm in diameter) polydimethylsiloxane (PDMS), and microgrooves (10μm, 30μm, and 50μm in width, and 2μm in depth) patterned PDMS, then imaged by fluorescent microscopy, time-lapse microscopy, and scanning electron microscopy (SEM). Cell morphology and migration were investigated through image analysis. The results suggest that the nanoporous and microgrooved surface structure induced totally different changes on BEL-7402 cells. Compared to the well-spread cells on the flat surface plate, the cells formed spheroids on the nanoporous AAM surface and nanoporous PDMS surface with no elongation and alignment, while the cells grew with elongated and aligned morphology along the microgrooves on the PDMS substrates. The BEL-7402 cell migration speed was significantly higher on the nanoporous substrates than on the flat surfaces. On the microgrooved PDMS substrates, the cells migrated along the groove direction and showed relatively small difference of the overall velocity compared to the cells on the flat PDMS surface. Our findings provide insights into the control of cell morphological features and migratory behaviour by using artificial nanoporous or microgrooved substrates, which can benefit the research on hepatocellular carcinoma metastasis, tissue engineering, and medical implant design.