Enriched Oxygen Mixtures in Athletes

Oxidative Stress Clinical Trial

— OXY-SPORT  

Official title:

Effects of Enriched Oxygen Mixtures and Exercise on Oxidative Stress and Stem Cells Proliferation in Athletes.

Clinical Trial Details — Status: Completed

Administrative data

• NCT number: NCT04366427
• Other study ID #: HEC-DSB/04-19
• Status: Completed
• Phase: Phase 2
• Start date: September 15, 2020
• Est. completion date: December 31, 2020

Study information

• Verified date: April 2021
• Source: University of Padova
• Contact: n/a
• Is FDA regulated: No
• Study type: Interventional

Clinical Trial Summary

Currently, Hyperbaric Oxigen (HBO) is a widely used treatment for several conditions. There are 14 indications for HBO, officially recognized by the Undersea and Hyperbaric Medical Society (UHMS), but research is discovering other interesting applications.

HBO plays an important role in enhancing antioxidant defense mechanisms by increasing radical oxygen species (ROS) and nitric oxide species (NOS). This controlled oxidative stress has been shown to stop the vicious circle of inflammation - damage - hypoxia already seen in several diseases. Increased neoangiogenesis has been demonstrated at pressures of 2 atmospheres absolute (ATA), while effects helping ischemic tissues need pressures between 2.5 and 2.8 ATA to develop. Also, stem cell proliferation and mobilization have been demonstrated after HBO treatments.

During sports activities, metabolism generates waste products - mostly CO2, lactic acid, but also ROS. HBO could be useful in modulating antioxidant mechanisms and increasing stem cell mobilization, thus helping cells in the recovery after training and sportive competitions.

The authors hypothesize that:

1. HBO can reduce oxidative stress and induce stem cell mobilization in healthy professional athletes;

2. hyperoxic mixtures can reduce oxidative stress and induce stem cells mobilization in healthy professional athletes;

3. HBO at low pressures (L-HBO at 1.45 ATA) is at least comparable to conventional HBO (at 2.5 ATA) in reducing oxidative stress and increasing stem cell mobilization.

The Authors will include healthy athletes. These will be randomly assigned to a control group, a L-HBO group, a HBO group, a 30% O2 group, or a 50% O2 group.

The Authors will assess oxidative stress changes and stem cells proliferation before and after 20 L-HBO/HBO/30% O2 mix/50% O2 mix treatments, and after 2 months after the end of treatments.

Description:

Subjects will be recruited through public announcements in local gyms and gathered to explain the protocol. Those willing to participate will sign a written informed consent and recruited. To be included, all the subjects will undergo a general medical screening to allow hyperbaric treatments. This will include weight, height, non-invasive arterial blood pressure, and heart rate measurements.

After inclusion, subjects will be randomly assigned to three arms using an electronic number generator by personnel not directly involved in the experiment:

• Arm 1(control): no intervention.

• Arm 2 (L-HBO): treated with oxygen at 1.45 ATA for 60 min (inclusive of compression and decompression times, and an air break of 3 minutes breathing air);

• Arm 3 (HBO): treated with oxygen at 2.5 ATA for 60 min (inclusive of compression and decompression times, and an air break of 3 minutes breathing air).

• Arm 4 (30% O2): breathing an air mixture with 30% of oxygen at atmospheric pressure (1 ATA).

• Arm 5 (50% O2): breathing an air mixture with 50% of oxygen at atmospheric pressure (1 ATA).

Subjects included in Arm 2, 3, 4, 5 will undergo a total of 20 treatments. They will follow a personalized diet proportional to their energetic expenditure.

The Authors will identify 3 time-points in the protocol:

TIME 0 (T0): immediately after inclusion, before any treatment or experiment; TIME 1 (T1): at the end of HBO treatments; TIME 2 (T2): 2 months after the end of HBO treatments.

The following exams will be performed on the included subjects:

• a standardized panel including Complete Blood Count (CBC), creatinine, Blood Urea Nitrogen (BUN), C reactive protein, and VES will be performed at T0, T1, and T2.

• oxidative stress markers will be analyzed on blood, urine, and saliva samples. On blood samples (T0; T1; T2), the Authors will measure IL-1 beta, IL-6, TNF-alfa, reactive oxygen species and total antioxidant capacity (by paramagnetic resonance), total (tot) and reduced (red) aminothiols (by fluorescence spectroscopy), 3-nitrotyrosine (3-NT) (by competitive immunoassay).

On urine samples (T0; T1; T2), the Authors will assess lipid peroxidation by measuring 8-isoprostane concentration (by competitive immunoassay), nitrite and nitrate (NO2/NO3) concentration (by colorimetry based on the Griess reaction), inducible Nitric Oxide Synthase (by ELISA commercially available kit), creatinine, neopterine, and uric acid concentrations, 8-oh-2-deoxyguanosine (by competitive immunoassay).

On saliva samples (T0; T1; T2) the Authors will measure reactive oxygen species and total antioxidant capacity (by paramagnetic resonance), and cortisol (by competitive immunoassay).

• stem cells will be analyzed on blood samples (at T0, T1, T2) (by flow cytometry).

Blood samples (approximately 6-12 ml) will be drawn from the veins of the forearms (preferentially on the non-dominant limb); plasma and erythrocytes will be separated by centrifuge at 1000×g for 10 min at 4°C. Urine samples will be collected by voluntary voiding in sterile containers. 1 mL of saliva will be obtained by Salivette devices (Sarstedt, Nümbrecht, Germany). The subjects will be instructed to refrain from drinking, eating, smoking, brushing their teeth, and using mouthwash in the 30 min before salivary collection.

All samples will be stored in multiple aliquots at - 80 °C until assayed and thawed only once before analysis.

With this setting, blinding of patients and investigators will be impossible due to different structural characteristics. However, outcome assessors will be blinded to patients' allocation.

Recruitment information / eligibility

• Status: Completed
• Enrollment: 42
• Est. completion date: December 31, 2020
• Est. primary completion date: November 15, 2020
• Accepts healthy volunteers: Accepts Healthy Volunteers
• Gender: All
• Age group: 18 Years to 35 Years

Eligibility

Inclusion Criteria:

• professional athletes

• performing at least 3 training sessions/week

Exclusion Criteria:

• previous pneumothorax

• problems with compensation maneuvers

• known epilepsy

• active smoker

Related Conditions & MeSH terms

• Oxidative Stress
• Stem Cell Research

Intervention

Combination Product:
• L-HBO
as previously described.
• HBO
as previously described.
• 30% O2
as previously described
• 50% O2
as previously described

Locations

Country	Name	City	State
Italy	Human Physiology Institute, Department of Biomedical Sciences, University of Padova	Padova	Veneto

Sponsors (2)

Lead Sponsor	Collaborator
University of Padova	Performa di Crocicchia Srl

Country where clinical trial is conducted

Italy, 

References & Publications (12)

Bosco G, Vezzani G, Mrakic Sposta S, Rizzato A, Enten G, Abou-Samra A, Malacrida S, Quartesan S, Vezzoli A, Camporesi E. Hyperbaric oxygen therapy ameliorates osteonecrosis in patients by modulating inflammation and oxidative stress. J Enzyme Inhib Med Chem. 2018 Dec;33(1):1501-1505. doi: 10.1080/14756366.2018.1485149. — View Citation

Bosco G, Yang ZJ, Di Tano G, Camporesi EM, Faralli F, Savini F, Landolfi A, Doria C, Fanò G. Effect of in-water oxygen prebreathing at different depths on decompression-induced bubble formation and platelet activation. J Appl Physiol (1985). 2010 May;108(5):1077-83. doi: 10.1152/japplphysiol.01058.2009. Epub 2010 Feb 25. — View Citation

Bosco G, Yang ZJ, Nandi J, Wang J, Chen C, Camporesi EM. Effects of hyperbaric oxygen on glucose, lactate, glycerol and anti-oxidant enzymes in the skeletal muscle of rats during ischaemia and reperfusion. Clin Exp Pharmacol Physiol. 2007 Jan-Feb;34(1-2):70-6. — View Citation

Camporesi EM, Bosco G. Mechanisms of action of hyperbaric oxygen therapy. Undersea Hyperb Med. 2014 May-Jun;41(3):247-52. Review. — View Citation

Fisher-Wellman K, Bloomer RJ. Acute exercise and oxidative stress: a 30 year history. Dyn Med. 2009 Jan 13;8:1. doi: 10.1186/1476-5918-8-1. — View Citation

Menzies P, Menzies C, McIntyre L, Paterson P, Wilson J, Kemi OJ. Blood lactate clearance during active recovery after an intense running bout depends on the intensity of the active recovery. J Sports Sci. 2010 Jul;28(9):975-82. doi: 10.1080/02640414.2010.481721. — View Citation

Morabito C, Bosco G, Pilla R, Corona C, Mancinelli R, Yang Z, Camporesi EM, Fanò G, Mariggiò MA. Effect of pre-breathing oxygen at different depth on oxidative status and calcium concentration in lymphocytes of scuba divers. Acta Physiol (Oxf). 2011 May;202(1):69-78. doi: 10.1111/j.1748-1716.2010.02247.x. Epub 2011 Mar 1. — View Citation

Moskowitz A, Andersen LW, Huang DT, Berg KM, Grossestreuer AV, Marik PE, Sherwin RL, Hou PC, Becker LB, Cocchi MN, Doshi P, Gong J, Sen A, Donnino MW. Ascorbic acid, corticosteroids, and thiamine in sepsis: a review of the biologic rationale and the present state of clinical evaluation. Crit Care. 2018 Oct 29;22(1):283. doi: 10.1186/s13054-018-2217-4. Review. — View Citation

Nasole E, Nicoletti C, Yang ZJ, Girelli A, Rubini A, Giuffreda F, Di Tano A, Camporesi E, Bosco G. Effects of alpha lipoic acid and its R+ enantiomer supplemented to hyperbaric oxygen therapy on interleukin-6, TNF-a and EGF production in chronic leg wound healing. J Enzyme Inhib Med Chem. 2014 Apr;29(2):297-302. doi: 10.3109/14756366.2012.759951. Epub 2013 Jan 30. — View Citation

Pedoto A, Nandi J, Yang ZJ, Wang J, Bosco G, Oler A, Hakim TS, Camporesi EM. Beneficial effect of hyperbaric oxygen pretreatment on lipopolysaccharide-induced shock in rats. Clin Exp Pharmacol Physiol. 2003 Jul;30(7):482-8. — View Citation

Van Hooren B, Peake JM. Do We Need a Cool-Down After Exercise? A Narrative Review of the Psychophysiological Effects and the Effects on Performance, Injuries and the Long-Term Adaptive Response. Sports Med. 2018 Jul;48(7):1575-1595. doi: 10.1007/s40279-018-0916-2. Review. — View Citation

Yang ZJ, Xie Y, Bosco GM, Chen C, Camporesi EM. Hyperbaric oxygenation alleviates MCAO-induced brain injury and reduces hydroxyl radical formation and glutamate release. Eur J Appl Physiol. 2010 Feb;108(3):513-22. doi: 10.1007/s00421-009-1229-9. Epub 2009 Oct 23. — View Citation

* Note: There are 12 references in all — Click here to view all references

Outcome

Type	Measure	Description	Time frame	Safety issue
Primary	Change in Reactive oxygen species production	Reactive oxygen species production (µmol min-1) (by paramagnetic resonance)	On blood and saliva: at baseline (T0), at the completion of treatments (Time 1: 5 weeks after the baseline) and 2 months after the end of treatments (Time 2)
Primary	Change in Total antioxidant capacity	Total antioxidant capacity (by paramagnetic resonance) (mM)	On blood and saliva: at baseline (T0), at the completion of treatments (Time 1: 5 weeks after the baseline) and 2 months after the end of treatments (Time 2)
Primary	Change in Cortisol levels	Cortisol (by competitive immunoassay) (ng/ml)	On saliva: at baseline (T0), at the completion of treatments (Time 1: 5 weeks after the baseline) and 2 months after the end of treatments (Time 2)
Primary	Change in nitrite and nitrate (NO2/NO3) concentration	nitrite and nitrate (NO2/NO3) concentration (by colorimetry based on the Griess reaction) (µM)	On urine: at baseline (T0), at the completion of treatments (Time 1: 5 weeks after the baseline) and 2 months after the end of treatments (Time 2)
Primary	Change in inducible Nitric Oxide Synthase (iNOS)	inducible Nitric Oxide Synthase (by ELISA commercially available kit) (IU mL-1)	On urine: at baseline (T0), at the completion of treatments (Time 1: 5 weeks after the baseline) and 2 months after the end of treatments (Time 2)
Primary	Change in aminothiols levels	total (tot) and reduced (red) aminothiols (by fluorescence spectroscopy) (µmol L-1)	On blood: Change from Baseline (T0) aminothiols concentration after the exercise test (Time 1: the day after baseline measurements), and at the completion of treatments after a second exercise test (Time 3: 5 weeks after the baseline)
Primary	Change in Cytokines levels	IL-1 beta, IL-6, TNF-alfa (pg ml-1)	On blood: at baseline (T0), at the completion of treatments (Time 1: 5 weeks after the baseline) and 2 months after the end of treatments (Time 2)
Primary	Change in lipid peroxidation markers	On urine samples, we will assess lipid peroxidation by measuring 8-isoprostane and 8-OH-deoxyguanosine concentration (by competitive immunoassay) - (pg mg-1 creatinine)	On urine: at baseline (T0), at the completion of treatments (Time 1: 5 weeks after the baseline) and 2 months after the end of treatments (Time 2)
Primary	Change in Renal damage markers	On urine samples, we will assess renal damage by measuring creatinine (g-L-1), neopterin (µmol·mol-1 creatinine), and uric acid levels (mg/dl).	On urine: at baseline (T0), at the completion of treatments (Time 1: 5 weeks after the baseline) and 2 months after the end of treatments (Time 2)
Primary	Change in 3-nitrotyrosine levels	3-nitrotyrosine (3-NT) (by competitive immunoassay)( nM·L-1)	On urine: at baseline (T0), at the completion of treatments (Time 1: 5 weeks after the baseline) and 2 months after the end of treatments (Time 2)
Primary	Change in Stem cells mobilization	Stem cells (by flow cytometry) (%)	On blood: at baseline (T0), at the completion of treatments (Time 1: 5 weeks after the baseline) and 2 months after the end of treatments (Time 2)