Despite the extensive research focused on the immunomodulatory properties of beta-glucans, alpha-glucans have received comparatively less attention. These molecules exhibit reduced immunogenicity compared to beta-glucans, a characteristic that could present valuable advantages in specific therapeutic contexts. This study evaluates the potential of three distinct extracted fractions (S_B, S_C, and R_D) obtained through different extraction methodologies from Coriolus versicolor, Pleurotus ostreatus, and Hericium erinaceus to enhance skin repair by promoting cellular proliferation, migration, and modulating immune responses. These fractions were previously characterized using several chemical and structural characterization methods that revealed significant content in bioactive molecules, particularly alpha glucans, exceeding 80% of total polysaccharide content. Cell viability was assessed using PrestoBlue and MTT assays in the HaCat cell line, revealing the non-toxicity of the compounds at the tested concentrations. Furthermore, proliferation assays (BrdU incorporation) and migration assays (scratch assay) in HaCaT cells were conducted at optimized concentrations (0.6 and 0.3 mg/mL). C. versicolor extracts (S_C and R_D) promoted the highest wound closure of the injured monolayer by 95% after 48 h (at 0.6 mg/mL) compared to 66% in the non-treated control, although BrdU incorporation revealed no significant changes in proliferation. In contrast, H. erinaceus extracts promoted increased cellular proliferation (>120% for S_B extracts at 0.6 mg/mL) with less impact in cell migration, suggesting selective enhancement of proliferation pathways. These differences may be attributed to variations in alpha-glucan structure and receptor interactions among the three species, leading to diverging effects on proliferation and migration. Additionally, immunomodulatory effects are being assessed by measuring key pro-inflammatory cytokines (TNF-α, IL1β, IL-8, and IL-6) and anti-inflammatory (IL 10) to elucidate how these polysaccharides regulate inflammation during healing and skin repair. These findings highlight the potential of alpha-glucans as multifunctional agents in dermatological applications, supporting their role in developing innovative, natural based therapies for skin regeneration.