A PMMA-based polymer previously shown to inhibit cell proliferation was compared to untreated PMMA. Conformation of adsorbed proteins, cell adhesion, cytoskeleton formation, and integrin activation were examined. Fibronectin adsorbed in a different conformation on the PMMA-based polymer exposing a different balance of the heparin-binding domains. Fibroblasts attached in equal numbers to both surfaces over a 4-h period, but the integrins involved in the adhesion process elicited different intracellular signaling pathways. Cells attached to PMMA showed activation of FAK and MAP as they spread using an assembled actin cytoskeleton. Cells attached to the polymer showed early and strong MAP activity that resulted in nonassembly of the actin cytoskeleton and sub-optimal cell spreading. We conclude that the chemistry of the polymer surface dictated a different conformation of the adsorbed proteins that resulted in alternative cell signaling and diminished cell spreading. This accounted for the biological inhibition previously reported on the PMMA-based polymer.