Regulation of endothelial cells migration on poly(D, L-lactic acid) films immobilized with collagen gradients

To investigate the effect of protein surface-density gradient on the motility of endothelial cells, we developed a novel approach for the fabrication of a collagen density gradient onto poly(d, l-lactic acid) (PDLLA) films in this study. The approach involves a sequential alkali hydrolysis of PDLLA films to produce a density gradient of -COOH moieties onto the films, which were activated and then covalently linked with collagen. A collagen surface-density gradient onto PDLLA films was thus generated by this approach. Contact angle measurement and confocal laser scanning microscopy (CLSM) were employed to confirm the formation of -COOH gradient and collagen gradient, respectively. All results proved the feasibility of the fabrication of a collagen density gradient onto PDLLA films via the approach. Endothelial cells cultured on the gradient areas with low and moderate collagen surface-densities displayed a strong motility tendency, with the values such as net displacement, total distance, chemotactic index, migration rate and cell trajectories in parallel to the gradient. However, endothelial cells grew on the gradient area with high collagen density demonstrated a reverse response to the collagen gradient clue. These results suggest that cell motility is regulated by the collagen gradient with a surface-density dependent manner. This study provides an alternative for the fabrication of protein surface-density gradient onto biodegradable substrates to investigate chemical stimuli induced cell directional motility. It is potentially important for understanding the controlled angiogenesis for implantation of tissue-engineered constructs.