Investigating the Impact of Hyperbaric Oxygen Therapy (HBOT) on Wound Healing

Wound Treatment Using HBOT

Acute and chronic wounds represent a major global healthcare challenge, contributing to significant morbidity and mortality. While the biology of wound healing has been extensively studied, optimizing therapeutic interventions remains a clinical priority. One widely used treatment is hyperbaric oxygen therapy (HBOT).

During HBOT, patients typically breathe 100% oxygen at ~2.4 atmospheres absolute (ATA) for 90 minutes, five times per week, for up to 40 sessions. This “supersaturate” the blood with oxygen, allowing delivery to poorly perfused tissues to meet the energy demands of cell proliferation and tissue repair required for wound healing. Cellular responses to HBOT include enhanced cell migration, increased collagen synthesis, neovascularization, and upregulation of growth factor production.

However, HBOT’s clinical effectiveness in chronic wound management, such as diabetic foot ulcers, has yielded mixed outcomes. The authors of our feature publication suggest this may be due to HBOT’s variable effects on the bioenergetic capacity of the cells involved in repair. Specifically, they sought to understand whether clinically-relevant HBOT treatment enhances or impairs mitochondrial function in wound-healing cells.

HBOT Effects on Cell Mitochondrial Dynamics during Wound Healing

Green and colleagues used high-resolution respirometry and fluorescence microscopy to quantify HBOT effects on mitochondrial function. To model wound-healing cells, Lifeline® Cell Technology’s primary human dermal fibroblasts, cultured in FibroLife media were exposed to a range of gas mixtures and hyperbaric pressures, conditions, as shown below. Cycle 3 most closely replicates clinical HBOT treatment. Baseline cells were kept in the chamber at ambient pressure with the same gas compositions described below for one hour.

Mitochondrial respiration, intermembrane potential, motility, dynamics, and the intracellular distribution of bioenergetic capacity were measured.

The authors found that HBOT exposure impacted multiple aspects of mitochondrial structure and function in the dermal fibroblasts. Mitochondrial size was generally stable between perinuclear and peripheral regions of the cells, except after Cycles 3 and 5, where peripheral mitochondria were significantly shorter. Compared to baseline, mitochondrial length decreased in the perinuclear region after Cycles 1–3 and in the peripheral region after all cycles except Cycle 4, suggesting that peripheral mitochondria are more susceptible to HBOT effects.

Mitochondrial motility was immediately suppressed in both regions following all HBOT conditions, with the largest decrease observed after Cycle 3. Although a compensatory increase in motility was observed after extended treatment (6hrs), ATP-linked respiration did not return to baseline levels, indicating that energy supply to key cellular regions was limited.

Mitochondrial intermembrane potential was preserved in peripheral mitochondria across all conditions but decreased significantly in perinuclear mitochondria after high-oxygen exposures (Cycles 3 and 5). The combination of reduced perinuclear motility and diminished intermembrane potential produces a transient imbalance between cellular energy demands and mitochondrial capacity, affecting critical wound-healing processes such as cell proliferation and migration.

Cellular respiration measurements revealed exposure-dependent effects. Basal ATP-linked respiration increased after Cycle 1, decreased after Cycles 2 and 3, and was unchanged in Cycles 4 and 5. Maximal respiration increased after Cycles 1, 3, 4, and 5, and spare respiratory capacity was elevated in all cycles, suggesting that HBOT can enhance the overall ability of cells to meet higher energy demands despite transient regional deficits.

These findings help explain the variable clinical outcomes of HBOT for wound treatment in chronic conditions such as diabetes or lower-extremity ischemia. While certain conditions (i.e., Cycle 1) create a mitochondrial profile that supports proliferation and migration, the clinically relevant protocol (Cycle 3: 100% O₂ at 2.4 ATA) actually suppresses mitochondrial motility and ATP-linked respiration in both perinuclear and peripheral regions. Therefore, although HBOT increases oxygen availability, it may temporarily reduce mitochondrial function and impair wound healing. Further studies are recommended using diabetes-related fibroblast models to identify more suitable HBOT treatments for chronic wound management as well as to understand how these mitochondrial responses influence HBOT efficacy.

Lifeline® Fibroblast Products  

Lifeline Cell Technology offers a comprehensive portfolio of primary human fibroblasts isolated from different tissues and specialized media optimized for their growth to help researchers investigate cellular function and biology in vitro including:

• Human Dermal Fibroblasts – Neonatal, Primary
• Normal Human Dermal Fibroblasts — Adult, Primary
• Human Dermal Fibroblasts – Neonatal, Xeno-Free, Primary
• Normal Human Primary Lung Fibroblasts
• Normal Human Bladder Fibroblasts
• Normal Human Vas Deferens Fibroblasts
• Normal Human Cardiac Fibroblasts
• Normal Human Uterine Fibroblasts
• Normal Human Gingival Fibroblasts
• Normal Human Scleral Fibroblasts
• Normal Human Prostate Fibroblasts
• FibroLife S2 Fibroblast Medium Complete Kit
• FibroLife Fibroblast Serum Free Medium Complete Kit

Join us next month for another installment of the Lifeline® blog to see how our cells and culture media are advancing biomedical research worldwide. If you have used our products in your publication, we’d love to feature your work here!