Polyethylene terephthalate (PET) is a major pollutant polymer, due to its wide use in food packaging and fiber production industries worldwide. Currently, there is great interest for recycling the huge amount of PET-based materials, derived especially from the food and textile industries. In this study, we applied the electrospinning technique to obtain nanostructured fibrillary membranes based on PET materials. Subsequently, the recycled PET networks were decorated with silver nanoparticles through the chemical reduction method for antimicrobial applications. After the characterization of the materials in terms of crystallinity, chemical bonding, and morphology, the effect against Gram-positive and Gram-negative bacteria, as well as fungal strains, was investigated. Furthermore, in vitro and in vivo biocompatibility tests were performed in order to open up potential biomedical applications, such as wound dressings or implant coatings. Silver-decorated fibers showed lower cytotoxicity and inflammatory effects and increased antibiofilm activity, thus highlighting the potential of these systems for antimicrobial purposes.