The global rise in antimicrobial resistance, particularly among ESKAPE pathogens, has intensified the demand for alternative therapeutic strategies. Silver nanoparticles (AgNPs) have exhibited broad-spectrum antimicrobial activity and represent a promising approach to combat multidrug-resistant infections. This study aimed to synthesize and functionalize AgNPs using various polymeric agents-ethylene glycol (EG), polyethylene glycol (PEG), polyvinylpyrrolidone (PVP), and their combinations-and to evaluate their antimicrobial and antibiofilm efficacy against clinically relevant bacterial strains. AgNPs were synthesized via chemical reduction and functionalized as Ag@EG, Ag@PEG, Ag@EG/PVP, and Ag@PEG/PVP. A total of 68 clinical isolates-including , , , , , and -were tested. Antimicrobial susceptibility was assessed using disc diffusion and broth microdilution assays, while antibiofilm activity was evaluated via the crystal violet method. Among all tested formulations, Ag@EG/PVP exhibited the highest antimicrobial and antibiofilm activity, with notably low minimum inhibitory concentrations (MIC) and minimum biofilm eradication concentrations (MBEC) for and . In contrast, AgNPs functionalized with PEG or EG alone showed limited efficacy. Biofilm-forming isolates, particularly spp., required higher concentrations for inhibition. These results highlight the critical role of functionalization in modulating the antimicrobial properties of AgNPs, with Ag@EG/PVP demonstrating potent activity against both planktonic and biofilm-associated multidrug-resistant bacteria. Overall, this study supports further developing AgNPs-based formulations as adjuncts or alternatives to conventional antibiotics, particularly for managing biofilm-related infections. Future research should focus on formulation optimization, safety assessment, and translational potential.