Bacterial biofilms affect many areas of human activity including food processing, transportation, public infrastructure, and most importantly healthcare. This study addresses the prevention of biofilms and shows that the surface charge of an abiotic substrate influences bacterial motility as well as the morphology and physiology of the biofilm. Grafting-from polymerisation was used to create polymer brush surfaces with different characteristics, and the development of Pseudomonas aeruginosa biofilms was followed using confocal microscopy. Interestingly, two types of biofilms developed on these surfaces: mushroom structures with high levels of cyclic diguanylate (c-di-GMP) were found on negatively charged poly (3-sulphopropylmethacrylate) (SPM) and zwitterionic poly (2-(methacryloyloxy)ethyl)dimethyl-3-sulphoproyl) ammonium hydroxide) (MEDSAH), while flat biofilms developed on glass, positively charged poly (2-(methacryloyloxy)-ethyl trimethyl ammonium chloride) (METAC), protein-repellent poly oligo(ethylene glycol methyl ether methacrylate) (POEGMA) and hydrophobic polymethylmethacrylate (PMMA). The results show that of all the surfaces studied, overall the negatively charged polymer brushes were most efficient in reducing bacterial adhesion and biofilm formation. However, the increased level of regulatory c-di-GMP in mushroom structures suggests that bacteria are capable of a quick physiological response when exposed to surfaces with varying physicochemical characteristics enabling some bacterial colonization also on negatively charged surfaces.