Dose-dependent effects of hyperbaric oxygen on PC12 cell viability and neurite outgrowth with and without NGF stimulation

Article Article scientifique Article Dose-dependent effects of hyperbaric oxygen on PC12 cell viability and neurite outgrowth with and without NGF stimulation eng 2025-10-03 2025-12 author André-Lévigne Dominik Division of Plastic, Reconstructive and Aesthetic Surgery, Geneva University Hospitals, Geneva, Switzerland. Electronic address: dominik@levigne.ch. 748833 author Maytain Margot Division of Plastic, Reconstructive and Aesthetic Surgery, Geneva University Hospitals, Geneva, Switzerland. 821191 author Pignel Rodrigue Diving and Hyperbaric Unit, Division of Emergency Medicine, Department of Anesthesiology, Clinical Pharmacology, Intensive Care and Emergency Medicine, Geneva University Hospitals and Faculty of Medicine, University of Geneva, Geneva, Switzerland. 818281 author Boet Sylvain 0000-0002-1679-818X Diving and Hyperbaric Unit, Division of Emergency Medicine, Department of Anesthesiology, Clinical Pharmacology, Intensive Care and Emergency Medicine, Geneva University Hospitals and Faculty of Medicine, University of Geneva, Geneva, Switzerland; Ottawa Hospital Research Institute, The Ottawa Hospital, University of Ottawa, Ottawa, ON, Canada. 413164 Département d'anesthésiologie, pharmacologie et soins intensifs (APSIC) author Kalbermatten Daniel Division of Plastic, Reconstructive and Aesthetic Surgery, Geneva University Hospitals, Geneva, Switzerland. 948183 Département de chirurgie (CHIRU) Daniel Felix Kalbermatten author Madduri Srinivas Division of Plastic, Reconstructive and Aesthetic Surgery, Geneva University Hospitals, Geneva, Switzerland. 948651 Département de chirurgie (CHIRU) 116-124 10.1016/j.bjps.2025.09.033 41151309 1748-6815 https://www.sciencedirect.com/science/article/pii/S1748681525005820 Hyperbaric oxygen therapy (HBOT) is a promising approach to promote peripheral nerve regeneration. However, the underlying mechanisms of action are poorly understood resulting in inconsistent treatment protocols and unclear indications. The present study evaluated the effects of single or repeated HBOT at different atmospheric pressures, with or without nerve growth factor (NGF) on PC12-cells. Cell proliferation, survival, and neurite outgrowth were analyzed. PC12-cells were exposed to a single session of HBOT or daily sessions over 4 consecutive days. Four groups were studied with or without NGF stimulation: control (21% oxygen at 1ATA (atmosphere absolute)) and 100% oxygen at 2.0ATA, 2.5ATA, or 3.0ATA. Brightfield imaging, live/dead assay, Alamar blue reagent, and tubulin-β-III immunostaining were performed. A single dose of HBOT enhanced proliferation of unstimulated PC12-cells under all experimental conditions compared to the control (p<0.05). In contrast, repeated HBOT increased dead cell count at 2.0ATA, 2.5ATA, and 3.0ATA compared to the control (p<0.01). Notably, repeated HBOT in NGF-stimulated PC12 cells resulted in maintained stable cell numbers and significant increase in axonal outgrowth (p<0.05). However, 3.0ATA impeded axonal outgrowth and induced cell death with or without NGF. Repeated HBOT decreased cell count and induced cell death in non-stimulated PC12-cells. NGF-stimulated cells showed stable cell numbers and no significant increase in cell death at 2.0 and 2.5ATA, whereas 2.5ATA significantly stimulated axonal outgrowth. Notably, 2.5ATA was the best pressure setting for neurite outgrowth stimulation in vitro. The data for nerve regeneration applications are promising but further research is needed to confirm the data in vivo and to investigate underlying mechanisms. Hyperbaric oxygen therapy Nerve growth factor PC12 cells Peripheral nerve regeneration Dewey 617 63 Département d'anesthésiologie, pharmacologie, soins intensifs et urgences 229 57 Département de chirurgie 222 false 1343 Reconstruction chirurgicale et régénération des tissus 1029 Journal of plastic, reconstructive & aesthetic surgery 111 72cd0cb9-7292-4ab5-8622-a2a47495fa21 Article (Published version) Public André-Lévigne_et_al._-_2025_-_Dose-.pdf 3775031 application/pdf CC BY-4.0 16119a2861518726d79065e2b4bd51374431716f002c0285073ce63da5b4dbd3 openAccess Faculté de médecine / Section de médecine clinique / Département d'anesthésiologie, pharmacologie, soins intensifs et urgences Faculté de médecine / Section de médecine clinique / Département de chirurgie

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Dose-dependent effects of hyperbaric oxygen on PC12 cell viability and neurite outgrowth with and without NGF stimulation	Journal of plastic, reconstructive & aesthetic surgery			2025	37	36

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Maytain, Margot

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