Development and Characterization of Sustainable Antimicrobial Food Packaging Films with Incorporated Silver Nanoparticles Synthesized from Olive Oil Mill By-Products

The growing accumulation of non-biodegradable petrochemical plastics and increasing food waste present urgent environmental and public health challenges. This study addresses both issues by developing biodegradable food packaging films from agar and starch, enhanced with antimicrobial properties by incorporating silver nanoparticles. The innovation of this work is the synthesis of novel agar–starch–silver nanoparticle coatings, where the contained nanoparticles were produced via green methods using two agro-industrial by-products of Greek olive oil production—olive stone extract and olive mill wastewater—as reducing agents. The morphology of the novel coatings was confirmed using transmission electron microscopy combined with energy-dispersive X-ray spectroscopy, revealing nanoscale particles with variable sizes. Additional film characterization was performed through Fourier-transform infrared spectroscopy, scanning electron microscopy coupled with energy-dispersive spectroscopy, and surface profilometry. Infrared spectroscopy analysis suggested the presence of functional groups responsible for nanoparticle stabilization, while energy-dispersive X-ray spectroscopy revealed silver aggregation in both olive stone extract and olive mill wastewater-derived films. Profilometry showed that films with olive mill wastewater-based nanoparticles had a rougher surface than those synthesized from olive stone extract. Antibacterial efficacy was tested against Escherichia coli (Gram-negative) and Staphylococcus epidermidis (Gram-positive) using a spot-on-film assay with high (10 6 CFU/film) and low (10 3 CFU/film) bacterial loads. After 72 h of incubation at 4 °C, both film types showed strong antibacterial activity at high bacterial concentrations, demonstrating their potential for active food packaging. These findings highlight a promising approach to sustainable food packaging within the circular economy, utilizing agricultural waste to create biodegradable materials with effective antimicrobial functionality.