Influence of surface energy distribution on neuritogenesis

PC12 cells are a useful model to study neuronal differentiation, as they can undergo terminal differentiation, typically when treated with nerve growth factor (NGF). In this study we investigated the influence of surface energy distribution on PC12 cell differentiation, by atomic force microscopy (AFM) and immunofluorescence. Glass surfaces were modified by chemisorption: an aminosilane, n-[3-(trimethoxysilyl)propyl]ethylendiamine (C8H22N2O3Si; EDA), was grafted by polycondensation. AFM analysis of substrate topography showed the presence of aggregates suggesting that the adsorption is heterogeneous, and generates local gradients in energy of adhesion. PC12 cells cultured on these modified glass surfaces developed neurites in absence of NGF treatment. In contrast, PC12 cells did not grow neurites when cultured in the absence of NGF on a relatively smooth surface such as poly-l-lysine substrate, where amine distribution is rather homogeneous. These results suggest that surface energy distribution, through cell-substrate interactions, triggers mechanisms that will drive PC12 cells to differentiate and to initiate neuritogenesis. We were able to create a controlled physical nano-structuration with local variations in surface energy that allowed the study of these parameters on neuritogenesis.