Systematic variation of microtopography, surface chemistry and elastic modulus and the state dependent effect on endothelial cell alignment.

We examined how variations in elastic modulus, surface chemistry and the height and spacing of micro-ridges interact and effect endothelial cell (EC) alignment. Specifically, we employed independent control of the surface properties in order to elucidate the relative importance of each factor. Polydimethylsiloxane elastomer (PDMSe) was fabricated with 1.5 or 5 microm tall, 5 microm spaced and 5, 10, or 20 microm wide ridge microtopographies. Elastic modulus was varied from 0.3, 1.0, 1.4, and 2.3 MPa by controlling oligomeric additives and crosslink density. Surface chemistry was left untreated, argon plasma treated, coated with fibronectin (Fn) or patterned with Fn tracks on flat PDMSe or the tops of micro-ridges. Primary porcine vascular ECs were cultured on the PDMSe substrates and nuclear form factor (NFF) was used to determine cell orientation relative to surface microtopography. Experimental results showed that microtopographical variation strongly altered EC alignment on Fn coated surfaces, but not on plasma treated surfaces. Interestingly, similar alignment was achieved with different orientation cues, either micropatterned chemistry (2D) or microtopography (3D). In total, the effect of varying one of the experimental parameters depended strongly on the state of the others, highlighting the need for multi-factor analysis of surface properties for applications where cells and tissue will contact synthetic materials.