Influence of Redox Stress on Crosstalk between Fibroblasts and Keratinocytes

SIMPLE SUMMARY: There has been significant scientific progress in skin care and skin damage repair, but the complete understanding of skin homeostasis is still beyond our reach. With an increase in environmental stress factors, the incidence rates of skin cancer and skin disorders are on the rise. Taken together with the incidence of scar- and burn-related morbidities, there is an urgent need to understand interactions between skin cells to develop novel therapies for the regeneration of healthy skin. One of the recurrent stress factors affecting the skin are the harmful free radicals, also referred to as oxidative stress. This study aimed to address the influence of oxidative stress on the interaction between two types of skin cells, keratinocytes and fibroblasts. The study utilized cold atmospheric plasma (CAP) to induce oxidative stress in cells and to assess the interactions between the two cell types. We showed that CAP can stimulate cells to enhance their proliferation and migration. This study provides a further understanding of skin cell regulation under stress conditions. Such knowledge may help in designing treatment therapies for rapid wound healing and skin repair. ABSTRACT: Although the skin is constantly subjected to endogenous and exogenous stress, it maintains a homeostatic state through wound repair and regeneration pathways. Treatment for skin diseases and injury requires a significant understanding of the various mechanisms and interactions that occur within skin cells. Keratinocytes and fibroblasts interact with each other and act as key players in the repair process. Although fibroblasts and keratinocytes are widely studied in wound healing and skin remodeling under different conditions, the influence of redox stress on keratinocyte-fibroblast crosstalk has not been thoroughly investigated. In this study, we used cold atmospheric plasma (CAP) to generate and deliver oxidative stress to keratinocytes and fibroblasts and to assess its impact on their interactions. To this end, we used a well-established in vitro 3D co-culture model imitating a realistic scenario. Our study shows that low CAP exposure is biocompatible and does not affect the viability or energetics of fibroblasts and keratinocytes. Exposure to low doses of CAP enhanced the proliferation rate of cells and stimulated the expression of key genes (KGF, MMP2, GMCSF, IL-6, and IL-8) in fibroblasts, indicating the activation and initiation of the skin repair process. Additionally, enhanced migration was observed under co-culture conditions under the given redox stress conditions, and expression of the upstream regulator and the effectors of the Hippo pathway (YAP and CYR61, respectively), which are associated with enhanced migration, were elevated. Overall, this study reinforces the application of CAP and redox stress in skin repair physiology.