Wound healing is a complicated process for maintaining skin integrity after injury, for which electrical stimulations (ES) have been attributed to promote wound healing by facilitating cell migration. Reducing the duration of the stimulation treatment from hours to minutes to promote efficient wound healing while avoiding cell damage poses a considerable challenge. Here, a promising technology of ultrashort pulse electric field (PEF), microsecond PEF at higher voltage, is proposed and realized to promote wound healing under a much short time (seconds) for the total treatment. We revealed that microsecond PEF regulated actin cytoskeleton reorganization and focal adhesion turnover, promoting fibroblasts migration in 2D cell cultures under the pulse stimulation. This accelerated fibroblast migration was accompanied by the mutual promotion with extracellular matrix (ECM) alignment in 3D microenvironments, which cooperatively benefit the eventual wound healing. These findings were further corroborated by the significant 2.5-fold enhancement observed in the healing of skin wounds in the classical mouse model by day 6 after electrical stimulation. Additionally, we coined an actin- and collagen-dependent mechanism of microsecond PEF-mediated wound healing. The quantitative mechanism proposed here for our novel microsecond pulse electric filed (μsPEF) methodology orients the new practical electric treatment toward a wide range of biomedical applications, such as wound healing, regenerative medicine and tissue engineering.