Antimicrobial food packaging was developed in response to the growing demand for longer fresh food shelf life and protection against foodborne infections. Therefore, the current study was conducted to investigate the antimicrobial effect of homemade biodegradable antimicrobial films incorporated with silver nanoparticles (Ag-NPs) against Staphylococcus aureus (S. aureus) experimentally inoculated in chicken breast. Chicken breast slices (5 × 5 cm) were experimentally inoculated with a cocktail of three S. aureus strains at a concentration of about 6.6 log10 CFU/cm2 and wrapped in homemade biodegradable antimicrobial films. Following that, the residual bacterial counts in wrapped chicken breast fillet slices were monitored for up to 10 days in a refrigerator (4 °C). It was found that at the end of the chilling period the films with biosynthesized Ag-NPs (1mM), biosynthesized Ag-NPs (2mM), chemically synthesized Ag-NPs (1mM), and chemically synthesized Ag-NPs (2mM) reduce the S. aureus counts by about 4.66, 5.24, 4.79, and 5.43 log10 CFU/cm2, respectively when compared with control films. Biodegradable antimicrobial films also prolonged the shelf-life of samples by approximately 4 days when compared to control samples.