Wound infections with rising incidences of multi-drug resistant bacteria are among the public health problems worldwide. The current study describes wound dressing materials made from biodegradable polyhydroxybutyrate (PHB) combined with AgNPs and gelatin (AgNPs/Gelatin/PHB). Microbial PHB was mixed with gelatin (1:2) to form a polymer matrix which was loaded with different concentrations of AgNPs (8.3–133 μg/mL). The statistical results of AgNPs synthesizing based on Box-Behnken design revealed that 1.247 mM silver nitrate and 24.054 % of Corchorus olitorius leaf extract concentration at pH (8.07) were the optimum values for the biosynthesis. UV–Vis spectroscopy, FTIR study and XRD reflects that nanoparticles are formed. The UV–Vis spectroscopy of Gelatin/PHB/AgNPs exhibited two specific bands at 298 nm and 371 nm, which confirm the formation of the conjugate. AgNPs had MICs and MBCs …