Finally, the long-lasting effect of the hydrogel loaded with vancomycin was tested, compared to vancomycin and E-coli as controls. The optical density at 600 nm was measured every 15 min for 20 h (Figure ). The vancomycin-loaded hydrogels showed a long-lasting antibacterial activity, and the hydrogel with the lowest concentration was the most effective. The empty hydrogels showed bacterial growth even above the E. coli control, with the lowestconcentration hydrogel having the highest bacterial growth. As shown in the photographs of the wounds (Figure ) and the percentage of wound contraction as determined by ImageJ software (Figure ), the Tegaderm and wound groups displayed slow healing during the 12-day period. In fact, the results demonstrate that the healing progress speed was much higher in the 8% hydrogel and 8% hydrogel+VAN groups: 3 days after surgery, the percentage of wound contraction is 46.12±15% for 8% hydrogel and 58.65±15.1 for 8% hydrogel+VAN, respectively. By contrast, these values were 37.2±16.2% and 36.3 ±18% for the Tegaderm and control groups, respectively. Moreover, Figure demonstrates the results of hematoxylin-eosin staining (H&E) staining of rats' wound tissues at day 5 and day 12 post-surgery. The results at day 5 displayed many inflammatory cells and disorganized micro-architectures in control groups compared to other groups. At day 12, the collagen fibers in 8% hydrogel and 8% hydrogel+VAN groups were more organized. In 8% hydrogel+VAN groups, a new layer of epidermis is observable. The histopathology results of H&E staining of the rat's main organs, including kidney, liver, and spleen (Figure ), did not show histopathological alterations such as dead tissue or alterations in the structure of cells. In addition, there was no signs of inflammation in 8% hydrogel, 8% hydrogel+VAN and Tegaderm groups compared to control group. As shown in the graphs, biochemical factors, including total protein (TP), albumin (ALB), calcium (Ca), blood urea nitrogen (BUN), creatinine (CREA) are not significantly different in 8% Hydrogel and 8% Hydrogel+VAN. Moreover, white blood cells (WBCs), red blood cells (RBCs), hemoglobin (HGB), hematocrit (HCT) and platelets (PLT), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), and mean corpuscular hemoglobin concentration (MCHC) are not significantly different among different groups.