Natural Killer (NK cells) are best known for their killing function in infection- and tumor-related responses, but also can shape immune responses involved in physiological processes, including wound healing. We recently reported that NK cells accumulate in skin wounds and express pro-inflammatory cytokines that may impede healing. Since impaired wound healing in diabetes is associated with persistent inflammation, we investigated whether NK cells contribute to impaired skin wound healing of diabetic mice. Our findings indicate that NK cells accumulate at higher levels in wounds in diabetic, and exhibit less mature phenotypes, compared to non-diabetic mice. In addition, local neutralization of CX3CL1 reduced NK cell accumulation in wounds of diabetic mice, suggesting that CX3CL1 plays a role in the infiltration/trafficking of these cells to the wound site. Finally, depletion of NK cells in diabetic wounds improved re-epithelization and collagen deposition, suggesting that the elevated levels of NK cells contribute to impaired healing associated with diabetes.