Enhanced Wound Healing Through Air‐Break Augmentation of Hyperbaric Oxygen Therapy Combined With Adipose‐Derived Stromal Cell Transplantation in a Murine Model

ABSTRACT

Hyperbaric oxygen therapy (HBOT) enhances wound healing by promoting angiogenesis and reducing hypoxia. However, the role of air-breaks—intermittent exposures to ambient air during HBOT—remains unclear. We investigated the effects of air-breaks on HBOT-mediated wound healing, particularly in combination with adipose-derived stromal cells (ASCs). Full-thickness wounds were created in C57BL/6 mice (n = 36) and assigned to control, HBOT (1 h/day, 2 ATA for 11 days), or HBOT with a 10-min air-break groups. In a second experiment, we evaluated ASC treatment combined with HBOT and air-breaks. Wound healing was assessed via gross examination, histology and gene expression analysis of collagen type 1 alpha 1 (Col1a1), hypoxia-inducible factor 1 alpha (Hif1a) and tumour necrosis factor (Tnf-α). Compared with HBOT alone, air-breaks significantly improved wound closure, epithelial regeneration and collagen deposition (p < 0.05). Gene analysis showed higher Col1a1 expression and lower Hif1a and Tnf-α levels in the air-break group. In ASC-treated wounds, air-breaks further accelerated healing, enhancing collagen synthesis and reducing hypoxia and inflammation. These findings suggest that incorporating air-breaks into HBOT protocols improves wound healing outcomes, both generally and in ASC-based therapies, by modulating collagen production, hypoxia and inflammation, and could optimise HBOT efficacy, particularly in cell-based regenerative therapies.