Effective therapies for skin wound healing and scar prevention are essential, and innovative approaches are necessary to address these clinical challenges. Due to their unique physicochemical properties, molybdenum trioxide nanoparticles (MoO 3 NPs) have emerged as promising candidates in wound healing and tissue repair. This study investigates the dose-responsive effects of MoO 3 NPs on dermal fibroblasts to provide insights into their potential applications in tissue repair and regeneration. Fibroblasts were exposed to a range of MoO 3 NP concentrations (from 50 to 400 μg/mL), and their cellular responses, including viability, proliferation, migration, and collagen synthesis, were assessed to comprehensively evaluate the influence of MoO 3 NPs on critical processes related to wound healing and fibrosis. Our findings reveal a dose-responsive relationship between MoO 3 NPs and skin fibroblast behavior; at lower concentrations (50 μg/mL), MoO 3 NPs exhibited stimulatory effects on fibroblast viability, proliferation, and migration, suggesting their potential as wound healing enhancers. Furthermore, this concentration stimulated the essential process of collagen synthesis, which plays a pivotal role in tissue repair. Nevertheless, with concentrations surpassing 50 μg/mL, a shift was noticeable as MoO 3 NPs started to impede cellular processes, particularly at high doses (400 μg/mL), resulting in decreased cell proliferation, migration, and angiogenesis. Also, the analysis of gene expression emphasized the regulatory influence of MoO 3 NPs on key genes associated with tissue repair and regeneration. Upregulation of transforming growth factor (TGF-β) and collagen genes was noted at lower concentrations, coinciding with improved healing responses, whereas higher doses downregulated these genes, potentially affecting fibroblast functionality. These findings emphasize the importance of precise nanoparticle dosing in therapeutic applications and provide valuable insights for the development of MoO 3 NP-based interventions to promote tissue repair.