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&#x2009;=&#x2009;36) and assigned to control, HBOT (1&#x2009;h/day, 2 ATA for 11&#x2009;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-&#x3b1;). Compared with HBOT alone, air-breaks significantly improved wound closure, epithelial regeneration and collagen deposition (p&#x2009;<&#x2009;0.05). Gene analysis showed higher Col1a1 expression and lower Hif1a and Tnf-&#x3b1; 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.