Diabetic wounds represent a major clinical challenge due to impaired healing processes such as reduced angiogenesis, chronic inflammation, and defective collagen remodeling. This study aimed to evaluate the therapeutic potential of a hyaluronic acid-based three-dimensional scaffold (HAS) in combination with hyperbaric oxygen therapy (HBOT) in promoting wound healing in a diabetic rat model. Diabetes was induced in Wistar rats, and full-thickness excisional wounds were created. Diabetic animals were divided into control, HBOT, HAS, and HAS+HBOT groups. Additionally, non-diabetic untreated rats (Healthy group) were considered as a control. Wound healing progression was assessed via wound contraction rate, stereological evaluations (fibroblast counts, neutrophil infiltration, and blood vessel density), cytokine profiling (VEGF, TGF-β, TNF-α, IL-1β), collagen deposition (Masson's trichrome, hydroxyproline assay), and biomechanical properties (maximum force and energy absorption). Wound contraction was significantly accelerated in all treatment groups compared to controls, with the HAS+HBOT group showing the greatest improvement (p < 0.05). Histological analysis revealed enhanced fibroblast proliferation and neovascularization, along with reduced neutrophil infiltration in treated groups, particularly in the HAS+HBOT group. Collagen content was markedly higher in treated wounds, supported by increased hydroxyproline levels and trichrome staining. The HAS+HBOT group also demonstrated elevated VEGF and TGF-β levels and decreased expression of TNF-α and IL-1β, indicating a favorable balance between regeneration and inflammation. Biomechanical testing confirmed superior maximum force and energy absorption in the HAS+HBOT group compared to all others. The combination of HAS and HBOT significantly enhances diabetic wound healing by promoting tissue regeneration, modulating inflammation, and restoring biomechanical integrity. This strategy holds promise as an effective therapeutic approach for chronic diabetic wounds.
