An adequate environment for wound healing improves cell proliferation and cell activity, rearranges fibroblasts, and suppresses the multiplication of bacteria. The aim of this study was to evaluate the wound-healing effects of a semi-occlusive hydrocolloid wound dressing (HWD) containing a topical antiseptic drug, benzalkonium chloride (BKC) in an infectious wound model. In vivo wound-healing profiles in rats exhibited that BKC-loaded HWD successfully promoted epithelialization rate, compared with gauze control or HWD without BKC. Furthermore, the significant decrease in desquamated epithelial regions, numbers of microvessels and inflammatory cells infiltrated in granulation tissues and granulation tissue areas were observed in the histopathological and histomorphometrical profiles of BKC-loaded HWD. In conclusion, these results suggest that BKC-loaded HWD facilitates wound healing and re-epithelization, and has promising effects for the treatment of infected wounds, as compared to HWD without BKC.