Cutaneous sulfur mustard (HD) exposure can lead to delayed wound healing. While animal models can be time-consuming and expensive, in vitro models offer a rapid and less expensive means to do initial screens of candidate therapies for prolonged wound healing. The purpose of this study was to develop an in vitro model to identify products with the potential for rapid fielding to effectively treat chemical warfare agent injuries and conventional wounds contaminated by chemical warfare agents. Human epidermal keratinocytes were seeded into 6 well plates and exposed to 0, 1, 10, or 100 μM HD. A 3 mm wide wound (disruption of the cell monolayer) was created using a sterile wounding instrument in each well. At various time points after wounding, wells were stained with 0.1% crystal violet to evaluate cell area within the wound. Images were captured and analyzed by computer-assisted histomorphometry to measure wound fill. After 5 days in culture, 0 and 1 μM HD-treated cells completely filled in the wound. In contrast, 10 and 100 μM HD-treated cells did not completely fill in the wound even after 12 days in culture. Additional wells were treated with a commercial wound cleanser or Pluronic F-68 surfactant following HD exposure and wound production. The wound cleanser killed the cells after just one dose. The surfactant-treated cells did not show an increase in wound healing over control cells. Other studies evaluated the use of growth factors to improve wound healing. Cells were treated with epidermal growth factor (EGF) or keratinocyte growth factor-2 (KGF-2). At higher concentrations, EGF did not appear to improve wound healing; however, at 1 ng/ml EGF appeared to improve wound healing over control levels. At the one concentration tested (100 ng/ml) and time point tested, KGF-2 did appear to improve wound healing. In conclusion, further studies will be conducted to evaluate possible products for improved wound healing. This in vitro wound healing model will be a useful tool to screen therapies for sulfur mustard-induced cutaneous injuries.