<p>Biofilms can be described as complex communities of microorganisms that adhere to surfaces and are embedded in a protective extracellular matrix, making them resistant to conventional treatments. Nanotechnology offers promising solutions for overcoming these challenges. This study introduces an innovative inorganic polymeric nanohybrid-based dressing, comprising a nanohybrid system (Cu-Ag-Co-silica) integrated into an electrospun cellulose acetate (CA) mat. The antibiofilm efficacy of this material was evaluated against a wide range of microbial strains, including both bacteria and fungi, in mono- and polymicrobial biofilms. The trimetallic silica nanohybrid-enhanced CA mat demonstrated superior antibiofilm activity, exhibiting the highest rate of radical scavenging activity (RSA) and an increased RSA with larger mat areas. Additionally, the material showed enhanced synergistic effects, significantly reducing minimum biofilm inhibitory concentration (MBIC₅₀), minimum killing time (MKT), and colony-forming units (CFU) over time compared to single-metal counterparts. Notably, filamentous fungi and gram-negative bacteria displayed reduced resilience to the antimicrobial dressing, while polymicrobial biofilms exhibited higher CFU counts than monomicrobial biofilms. These findings highlight the potential of this innovative inorganic polymeric nanohybrid-based dressing as a broad-spectrum agent for combating a wide variety of microbes in chronic wounds. This breakthrough offers new insights into antibiofilm mechanisms and practical strategies for effectively addressing biofilm-related infections.</p>