To better understand and optimize the fine interactions that occur during adhesion events between human cells and synthetic materials, we seeded human umbilical vein endothelial cells (HUVEC) onto ethylene-vinyl alcohol (EVOH) copolymer films prepared by casting. Different adhesive proteins, e.g. fibronectin and gelatin, and the monoclonal antibody (MoAb) CLB-HEC19 specific for the endothelial cell membrane were used to coat the materials. We used atomic force microscopy (AFM) to analyse the EVOH film structure, to test its planarity and homogeneity, before seeding it with endothelial cells. The metabolic changes induced in the endothelial cells by interactions with the copolymer functional groups and the adhesive proteins were monitored by a micro-electronic pH sensor, positioned close to the HUVEC monolayer. We found that the adhesion of HUVEC onto various substrates was finely modulated by the MoAb CLB-HEC19 and that the endothelial cell metabolic rate was enhanced when cultured onto a CLB-HEC19 coating. Surface roughness seems also to play a role in the interaction with HUVEC. The AFM measurement analysis demonstrated that L6 surface is rougher than R20. These surface characteristics could favour cell adhesion; in fact HUVEC adhesion results on R20 were significantly lower than on L6.