If untreated, skin wounds can lead to severe complications. Depending on the type of injury, long-term antibiotic administration is often required, and this decreases patient compliance. This limitation could be addressed by applying dressings capable of preventing infections by controlling drug release to the wound site. In this research, biodegradable wound dressings were investigated, based on natural polymers chitosan and alginate and incorporating the broad-spectrum gentamicin as antibiotic. Specifically, gentamicin was loaded into alginate nanoparticles, which were then loaded into chitosan-based films. This approach aimed at obtaining a system capable of modulating antibiotic release. The obtained nanoparticles had an average diameter of 86 nm and polydispersity index of 0.15. Antibiotic loading was around 600 µg/mg, with loading efficiency close to 100%. Films incorporating nanoparticles were compared to control films, which contained only gentamicin. Results showed that nanoparticles incorporation decreased film's swelling in phosphate buffer saline, thus leading to a decrease in burst release while cytocompatibility for human dermal fibroblasts was maintained. Antibacterial activity was confirmed against both gram-positive and gram-negative bacteria. Moreover, the antibiotic was released as a function of pH, with distinct behavior at pHs ranging from 7.4 to 5.5. This indicates that alginate nanoparticles dispersed in chitosan films effectively release gentamicin on demand.