17β-Estradiol Treatment Improves Wound-Healing–Related Effects in an In-Vitro Fibroblast–Endothelial Co-culture Model

Introduction: Estrogen plays a central role in regulating multiple phases of wound healing, but its direct influence on cooperative interactions between fibroblasts and endothelial cells is not well established. This study investigated whether 17β-estradiol (E2) enhances wound-healing–related cellular behaviors in a fibroblast–endothelial co-culture model. Methods: Human dermal fibroblasts (C163) and HUVECs were co-cultured at a 1:1 ratio and treated with E2 (0, 50, 75, or 100 ng/mL). Cell viability (MTT assay), collective migration (scratch assay), and expression of Vimentin and CD31 (qPCR using Transwell co-culture and immunofluorescence) were evaluated. Results: E2 significantly increased the viability and migration of both cell types in a dose-dependent manner, with the strongest effect at 75 ng/mL. VIM and CD31 transcript levels were similarly upregulated at all concentrations, with peak expression at 75 ng/mL (p < 0.05). Immunofluorescence confirmed enhanced Vimentin and CD31 protein expression. Preliminary monoculture observations demonstrated a weaker E2 response compared with co-culture, indicating that fibroblast–endothelial interaction amplifies hormone-mediated effects. discussion: The co-culture approach better reflects in vivo wound environments compared to single-cell models, highlighting estrogen’s coordinated effects on multiple cell types. However, this model does not include other relevant cells such as keratinocytes or immune cells, and the mechanisms underlying the observed responses were not explored here. Further studies should investigate signaling pathways and optimal dosing to guide potential therapeutic applications. Discussion: Our study found that E2 improves fibroblast–endothelial cooperation by increasing cell survival, migration, and expression of Vimentin and CD31 in a co-culture paradigm. The highest effects were observed at 75 ng/mL, indicating an appropriate estrogen dose for coordinated wound-healing responses. The lower response in monoculture emphasizes the necessity of intercellular communication in mediating E2 effects. Overall, E2 appears to support important cellular processes involved in wound healing. Conclusion: 17β-Estradiol promotes coordinated fibroblast–endothelial activity by enhancing proliferation, migration, and cytoskeletal/angiogenic marker expression. These findings support the potential relevance of E2 as a modulator of key processes involved in wound repair.