Wound healing is a complex process regulated by various immune and biological systems. Keratinocytes are crucial cell type in the skin and wound healing. They play an important role in the healing process by rapidly covering the surfaces of skin and wounds, thereby resetting the epithelial barrier. Wound healing is important for diseases such as diabetes and burns and the importance of wound treatment methods for these diseases continues to increase. During the initial phases of wound healing, it is recognized that the activation of nitric oxide synthases (NOS), particularly inducible nitric oxide synthases (iNOS) by inflammatory cells, where and macrophages plays an important role. It is known that reactive oxygen species (ROS) regulate wound contraction, but its exact mechanism remains unclear. In this study, we investigated the impact of human keratinocytes treated with a natural product-derived nitric oxide (NO) on wound healing. The effects of NO on migration abilitiy of keratinocytes were assessed through wound healing assay, transwell migration assay, FACS analysis, reverse transcrption (RT)-PCR and western blot. According to the research findings, the selected NO donor fucoidan and Stewartia pseudocamellia (S. pseudocamellia) root methanol hot water extract (MHWE) exhibited a dose-dependent enhance in wound confluence when treat to human keratinocytes (HaCaT) cells with scratch induced wounds as compared to the control group. Furthermore, in the transwell migration assay, it was observed a dose dependent increase in the number of migrated cells compared to the control group. To determine whether NO promotes intracellular ROS, ROS levels were measured through FACS analysis. This analysis confirmed the activation of ROS generation. The expression levels of migration- related genes and proteins were measured by RT-PCR and western blot. The results revealed a dose- dependent manner increase in the exprssion levels of genes and proteins. In conclusion, the amalgamation of these research findings suggests that NO could exert a beneficial influence on wound healing. This suggests the potential for NO to be considered as an effective therapeutic apporoach in the development of strategies for optimal wound healing.