Biocompatible smart interfaces play a crucial role in biomedical or tissue engineering applications, where their ability to actively change their conformation or physico-chemical properties permits finely tuning their surface attributes. Polyelectrolytes, such as acrylic acid, are a particular type of smart polymers that present pH responsiveness. This work aims to fabricate stable hydrogel films with reversible pH responsiveness that could spontaneously form wrinkled surface patterns. For this purpose, the photosensitive reaction mixtures were deposited via spin-coating over functionalized glasses. Following vacuum, UV, or either plasma treatments, it is possible to spontaneously form wrinkles, which could increase cell adherence. The pH responsiveness of the material was evaluated, observing an abrupt variation in the film thickness as a function of the environmental pH. Moreover, the presence of the carboxylic acid functional groups at the interface was evidenced by analyzing the adsorption/desorption capacity using methylene blue as a cationic dye model. The results demonstrated that increasing the acrylic acid in the microwrinkled hydrogel effectively improved the adsorption and release capacity and the ability of the carboxylic groups to establish ionic interactions with methylene blue. Finally, the role of the acrylic acid groups and the surface topography (smooth or wrinkled) on the final antibacterial properties were investigated, demonstrating their efficacy against both gram-positive and gram-negative bacteria model strains ( and ). According to our findings, microwrinkled hydrogels presented excellent antibacterial properties improving the results obtained for planar (smooth) hydrogels.