The stiffness of elastomeric surfaces influences the mechanical properties of endothelial cells

Abstract: Optimal characterization of the mechanical properties of both cells and their surrounding is an issue of major interest. Indeed, cell function and development are strongly influenced by external stimuli. Furthermore, a change in cell mechanics might, in some cases, associate with diseases or malfunctioning. In this work, atomic force microscopy (AFM) was applied to examine the mechanical properties of the silicone elastomer polydimethylsiloxane (PDMS) a common substrate in cell culture. Force spectroscopy analysis was done over different specimens of this elastomeric material containing varying ratios of resin to cross-linker in its structure (5:1, 10:1, 20:1, 30:1 and 50:1), which impacts the final material properties (e.g., stiffness, elasticity). To quantify the mechanical properties of the PDMS, factors as the modulus of Young, the maximum adhesive forces as well as both relaxation amplitudes and times upon constant height contact of the tip (dwell time different of zero) were calculated from the different segments forming the force curves. It is demonstrated that the material stiffness is increased by prior oxygen plasma treatment of the sample, required for hydrophilic switching, contrarily to what observed for its adhesiveness. Subsequent incubation of endothelial HUVEC cells on top of these plasma treated PDMS systems yields minor variation in cell mechanics in comparison to those obtained on a glass reference, on which cells show much higher spreading tendency and, by extension, a remarkable membrane hardening. Thus, surface wettability turns a factor of higher relevance than substrate stiffness inducing variations in the cell mechanics.