The mechanisms underlying the complex and multistage wound-healing process are not yet completely understood. One of the most important and intriguing questions remaining is the effect of the interactions between wounds and the microflora that are present in wounds. In this report, we describe the first study of the effect of treating murine skin wounds with topical bacterial lipopolysaccharide (LPS), the main exogenous ligand of Toll-like receptor 4. Our findings demonstrate that LPS treatment strongly affects the wound-healing process by accelerating the resolution of inflammation, increasing macrophage infiltration, enhancing collagen synthesis, and altering the secretion of a number of mediators that are involved in the skin regeneration process. Topical LPS treatment upregulated the secretion of proinflammatory cytokines [interleukin (IL)-6, IL-1β, and leukemia inhibitory factor (LIF)] and CC-chemokines (CCL2/MCP-1, CCL7/MCP-3, CCL3/MIP-1α, and CCL5/RANTES), but not CXC-chemokines (CXCL2/MIP-2 and CXCL9/MIG). The secretion of growth factors (vascular endothelial growth factor, transforming growth factor-β1 (TGF-β1), and fibroblast growth factor 2) at the wound site was also upregulated. Taken together, these results suggest that the topical application of LPS at the wound surface affects the inflammatory process and promotes the wound healing of injured skin.