the current study aimed to formulate Selenium-chitosan-Mupirocin (M-Senps-ccH) complex. the nanohybrid system was prepared using chitosan-cetyltrimethylammonium bromide (ctAB)based hydrogel (ccH) that entrapped mupirocin (M) and selenium nanoparticles (Senps). the in vitro studies were performed by evaluation of the antibacterial activity and toxicity on L929 mouse fibroblast cell line. The in vivo study was conducted on rat diabetic wound infection model that was infected by mupirocin-methicillin-resistant Staphylococcus aureus (MMRSA). the wounds were treated by M-SeNPs-CCH nanohybrid system with concentrations of M; 20 mg/ml, CCH; 2 mg/ml and SeNPs; 512 μg/ml in two times/day for 21 days. The therapeutic effect of this nanohybrid system was evaluated by monitoring wound contraction and histopathological changes. evaluation of the average wound healing time showed a significant difference between the treatment and control groups (P≤0.05). The histopathological study indicated that the amount of wound healing was considerable in M-Senps-ccH nanohybrid system groups compared to the control and M groups. the M-Senps-CCH nanohybrid system formulated in this study was able to reduce 3-fold MIC of mupirocin with synergistic antibacterial activity as well as to play a significant role in wound contraction, angiogenesis, fibroblastosis, collagenesis, proliferation of hair follicle, and epidermis growth compared to the control group (P ≤ 0.05). This research suggests that this nanohybrid system might be a development for the treatment of diabetic wound infection at mild stage. Management of the infectious diabetic foot ulcer remains a global health issue 1,2. According to the World Health Organization (WHO), about 422 million people worldwide suffer from diabetes 2. About 25% of diabetic patients experience diabetic foot wounds in their lifetime 3,4 , half of them suffer infection 3. Diabetic wounds are classified into three groups, i.e. mild, moderate and severe 3. To avoid amputation and reduce health care costs, it is essential to control the infection 5. Gram-positive bacteria such as Staphylococcus aureus and beta-hemolytic streptococci are common infectious agents in the mild stage of diabetic foot wounds 3,5. Regarding the increasing incidence of antibiotic-resistant strains such as methicillin-resistant S. aureus (MRSA) 6,7 , the treatment of infection caused by this organism has