Abstract Background and aim: Treatment of burn wound infections has become a global challenge due to the spread of multidrug-resistant bacteria; therefore, the development of new treatment options for the mentioned infections is essential. Platelets have drawn much attention for this purpose because they are a safe and cost-efective source of diferent antimicrobial peptides and growth factors. The present study evaluated antibacterial efects and wound healing properties of Platelet-derived Biomaterial (PdB) against Acinetobacter baumannii and Klebsiella pneumoniae burn wound infections. Methods: PdB was prepared through the freezing and thawing process and then, in vitro antibacterial efect was determined by disk difusion and broth microdilution methods. Afterward, burn wound was inficted on 56 rats, infected with both bacteria, and topical administration was performed to evaluate antibacterial efects and wound healing properties of PdB. Results: In vitro results showed that PdB inhibited the growth of A. baumannii in the highest dose (0.5), while we did not detect any inhibitory efects against K. pneumoniae. By contrast, PdB signifcantly inhibited the growth of bacteria in treated animal wounds compared to the control groups (P value<0.05). Macroscopic assessments pointed to the signifcant enhancement of wound closure in the treated animals. In addition, histopathological examination demonstrated that treatment of rats with PdB led to a considerable increase in re-epithelialization and attenuated the formation of granulation tissue (P value<0.05). Conclusion: The use of topical PdB is an attractive strategy for treating A. baumannii and K. pneumoniae burn wound infections because it inhibits bacterial growth and promotes wound healing properties.