Molecular Hydrogen Inhalation: Effects on Health, Exercise Capacity and Inflammatory Response - in Vivo/in Vitro Studies

NCT07130942 | Not Yet Recruiting

• 1 other identifier: AWFiS/2025_8_JM
• Study Type: interventional
• Target: 250
• Locations: 1 country
• Sites: 1

Brief Summary

Summary To comprehensively address the research aims and explore the state of knowledge of the mechanisms of biochemical response to specific tissue affecting method in form of a molecular hydrogen inhalation and will allow to determine its role on the post-exercise response in form of changes in biochemical markers secretion, expression of selected trophic factors and changes in iron metabolism in this process. Moreover, this project will try to take into account the role of hepcidin, vitamin D and cfDNA. Additionally, the present project may contribute to the determination of the role of presented inhalation procedure on cells proliferation, as example of anty-tumor proprieties, regulation of the expression of genes related to the stress response (HSF-1, NF-kB, TNF-dependent pathway), muscle cell growth (e.g. myostatin gene), energy pathways (e.g. GAPDH, LDH) and the membrane transport and the hedgehog pathway ls (including Gli1), Hif-1-alpha and NF-kB. 1.1. Primary Objectives

• For the measurement of Aerobic Components of Fitness and post-aerobic exercises response Bruce Treadmill Test will be performed.
• The blood collection for diagnostic tests will be strictly dependent on the requirements of a particular designation,

Conditions

Physiological Adaptations; Sport Performance; Aerobic Capacity; Anaerobic Power; Sport Recovery

Keywords

molecular hydrogen; anti-cancer potential; inflammatory response; exercise capacity; inhalation

Outcome Measures

Primary Outcomes (4)

• Measurement of the inflammation blood markers using Magnetic Luminex Performance Assays

  The intake will be performed in appropriate standardized BD Vacutainer tubes by qualified medical personnel: Serum and plasma will be separated from the samples, Each sample to obtain the serum will be centrifuged at 2500-3000 rpm for 10 minutes in 4° C and stored in 1.5 ml tubes at - 80° C until the assay (no longer than 6 months). The following secretory factors and markers of inflammation will be determined in serum or blood plasma (depending on the requirements of the method used) and then analysed in detail using immu: • high sensitive C-protein (hsCRP), • creatine kinas (CK), • lactic acid (LA), • interleukin 6 (IL6), • interleukin 10 (IL10), • interleukin 15 (IL15),

  Change from baseline at 24 hours after fatigue-induced exercise (before and after each protocol)

• Measurement of the Iron metabolism blood markers using Magnetic Luminex Performance Assays and ELISA Assays

  The intake will be performed in appropriate standardized BD Vacutainer tubes by qualified medical personnel: Serum and plasma will be separated from the samples, Each sample to obtain the serum will be centrifuged at 2500-3000 rpm for 10 minutes in 4° C and stored in 1.5 ml tubes at - 80° C until the assay (no longer than 6 months). The following secretory factors and markers will be messured: • free Iron • hepcidin, • transferrin, • ferritin, • EPO, • EFRE, • TIBC,

  Change from baseline at 24 hours after fatigue-induced exercise (before and after each protocol)

• Measurement of neurotrophic, angiogenic blood markers using Automated Hematology Analyzers and Magnetic Luminex Performance Assays

  The intake will be performed in appropriate standardized BD Vacutainer tubes by qualified medical personnel: Serum and plasma will be separated from the samples, Each sample to obtain the serum will be centrifuged at 2500-3000 rpm for 10 minutes in 4° C and stored in 1.5 ml tubes at - 80° C until the assay (no longer than 6 months). The following secretory factors and markers of inflammation will be determined: angiogenin (ANG), • insulin-like growth factor-1 (IGF-1), • Growth Differentiation Factor 15 (GDF15), • nerve growth factor (NGF), • Amyloid Precursor Protein- α (sAPPα), • angiopoietin (ANGP1), • brain-derived neurotrophic factor (BDNF) • growth differentiation factor 15 (GDF-15), • tumor necrosis factor α (TNFα).

  Change from baseline at 24 hours after fatigue-induced exercise (before and after each protocol)

• The general assessment of the homoeostasis Automated Hematology Analyzers

  • blood morphology, • glycemia, • free iron, • activity of alanine aminotransferase (ALT), • aspartate aminotransferase (AST), • creatine kinase (CK), • lactate dehydrogenase (LDH), • concentration of insulin, • total cholesterole • creatinine,

  Before all procedures and before post-hydrogen inhalation testing

Secondary Outcomes (3)

• Measurement of proteins involved in antioxidant defence using Western Blotting Techniques

  Before and after 1-time and 14-times inhalation using a molecular hydrogen generator

• Messurment of cfDNA changes

  Before, and 5 min, 60 min after every WAnT and Bruce Test performance

• Measurement of isolated serum cytotoxic activity

  Up to 12 months post-experimental period

Other Outcomes (1)

• Body composition analyses using bioelectrical impedance analysis (BIA)

  During the initial visit, and before and after every experimental phase (1-time; 14-times inhalation using a molecular hydrogen generator)

Study Arms (4)

1-time molecular hydrogen inhalation intervention_SHAM intervention

SHAM COMPARATOR

1 time SHAM intervention

Procedure: Inhalation using a molecular hydrogen generator

Two weeks molecular hydrogen inhalation intervention_SHAM intervention

SHAM COMPARATOR

14-days molecular hydrogen inhalation intervention

Procedure: Inhalation using a molecular hydrogen generator

1-time molecular hydrogen inhalation intervention

EXPERIMENTAL

1-time molecular hydrogen inhalation intervention

Procedure: Inhalation using a molecular hydrogen generator

Two weeks molecular hydrogen inhalation intervention

EXPERIMENTAL

14-days molecular hydrogen inhalation intervention

Procedure: Inhalation using a molecular hydrogen generator

Interventions

Inhalation using a molecular hydrogen generator PROCEDURE

Hydrogen inhalation will be performed by using a hydrogen gas generator similar in terms of capabilities to the generator Hycellvator ET100 (Helix Japan, Co., Ltd., Tokyo, Japan). The apparatus will be generating 30.0 mL/s gas mixture, consisting of 68.0% hydrogen (hydrogen purity, 99.99%) and 32.0% of oxygen. All gases will be supplied through a nasal cannula connected to the gas generators. Although we could not measure directly the hydrogen and oxygen An average inspiratory flow rate will be adjusted to 500 mL/s at rest, the hydrogen concentration in the inspired gas must have been around 4.08% at most. Control population will have inhalation with the use of gas generator that has the same outer shape as the used generator to produce Placebo (30.0 mL/s, ambient air 400 m above sea level) consisting of 0.00005% of hydrogen and 20.9% of oxygen.

Also known as: molecular hydrogen inhalation, molecular hydrogen therapy, Molecular hydrogen (H2) therapy

1-time molecular hydrogen inhalation intervention; 1-time molecular hydrogen inhalation intervention_SHAM intervention; Two weeks molecular hydrogen inhalation intervention; Two weeks molecular hydrogen inhalation intervention_SHAM intervention

Eligibility Criteria

• Age: 18 Years - 49 Years
• Sex: all
• Healthy Volunteers: Yes
• Age Groups: Adult (18-64)

You may qualify if:

• The study will be used purposeful selection of the following criteria:
• age 18-25,
• not taking medicines during the study,
• negative history of cardiovascular disorders,
• negative history of autonomic nervous system disorders,
• negative history of mental disorders,
• negative history of cerebrospinal traumas,
• negative history of other diseases that may directly affect obtained results,
• good health status (no concurrent injuries),
• no drugs intake,
• no supplements consumption,
• In the study as a not training grope age and morphologically appropriate participation will take part. Participants will be recruited basing on a voluntary letter of intent. All representatives of the analysed group participating in the pre-qualification research will fill in the physical activity sheet - Global Health Activity Questionnaire - World Health Organization in Polish adaptation. This will allow to eliminate people who report high levels of physical activity (similar to the level of sport training individuals).

You may not qualify if:

• taking medicines during the study,
• history of cardiovascular disorders,
• history of autonomic nervous system disorders,
• history of mental disorders,
• history of cerebrospinal traumas,
• history of other diseases that may directly affect obtained results,
• concurrent injuries,
• drugs intake,
• supplements consumption.

Sponsors & Collaborators

• Gdansk University of Physical Education and Sport: lead
• Medical University of Gdansk: collaborator
• Kazimierz Wielki University: collaborator

Study Sites (1)

University of Physical Education and Sport (GUPES)

Gdansk, Pomeranian Voivodeship, 80-336, Poland

Location

Related Publications (7)

• Chomczynski P, Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156-9. doi: 10.1006/abio.1987.9999.

  PMID: 2440339 BACKGROUND

• Sim M, Kim CS, Shon WJ, Lee YK, Choi EY, Shin DM. Hydrogen-rich water reduces inflammatory responses and prevents apoptosis of peripheral blood cells in healthy adults: a randomized, double-blind, controlled trial. Sci Rep. 2020 Jul 22;10(1):12130. doi: 10.1038/s41598-020-68930-2.

  PMID: 32699287 BACKGROUND

• Farley JB, Stein J, Keogh JWL, Woods CT, Milne N. The Relationship Between Physical Fitness Qualities and Sport-Specific Technical Skills in Female, Team-Based Ball Players: A Systematic Review. Sports Med Open. 2020 Apr 15;6(1):18. doi: 10.1186/s40798-020-00245-y.

  PMID: 32297147 BACKGROUND

• Timon R, Olcina G, Gonzalez-Custodio A, Camacho-Cardenosa M, Camacho-Cardenosa A, Martinez Guardado I. Effects of 7-day intake of hydrogen-rich water on physical performance of trained and untrained subjects. Biol Sport. 2021 Jun;38(2):269-275. doi: 10.5114/biolsport.2020.98625. Epub 2020 Oct 22.

  PMID: 34079172 BACKGROUND

• Botek M, Khanna D, Krejci J, Valenta M, McKune A, Sladeckova B, Klimesova I. Molecular Hydrogen Mitigates Performance Decrement during Repeated Sprints in Professional Soccer Players. Nutrients. 2022 Jan 25;14(3):508. doi: 10.3390/nu14030508.

  PMID: 35276867 BACKGROUND

• Itoh T, Hamada N, Terazawa R, Ito M, Ohno K, Ichihara M, Nozawa Y, Ito M. Molecular hydrogen inhibits lipopolysaccharide/interferon gamma-induced nitric oxide production through modulation of signal transduction in macrophages. Biochem Biophys Res Commun. 2011 Jul 22;411(1):143-9. doi: 10.1016/j.bbrc.2011.06.116. Epub 2011 Jun 23.

  PMID: 21723254 BACKGROUND

• Hancock JT, Russell G. Downstream Signalling from Molecular Hydrogen. Plants (Basel). 2021 Feb 14;10(2):367. doi: 10.3390/plants10020367.

  PMID: 33672953 BACKGROUND

MeSH Terms

Conditions

Respiratory Aspiration

Interventions

Hydrogen; Therapeutics

Condition Hierarchy (Ancestors)

Respiration Disorders; Respiratory Tract Diseases; Pathologic Processes; Pathological Conditions, Signs and Symptoms

Intervention Hierarchy (Ancestors)

Elements; Inorganic Chemicals; Gases

Study Officials

• Jędrzej Antosiewicz, Full Profesor

  Medical University of Gdańsk, Department of Bioenergetics and Physiology of Exercise, Gdańsk, Poland

  PRINCIPAL INVESTIGATOR

Central Study Contacts

Jan Pawel Mieszkowski JP Mieszkowski, PhD hab, PhD hab

CONTACT

+48 58 554 71 21; jan.mieszkowski@awf.gda.pl

Andrzej Kochanowicz, PhD hab.

CONTACT

andrzej.kochanowicz@awf.gda; andrzej.kochanowicz@awf.gda.pl

Study Design

• Study Type: interventional
• Phase: not applicable
• Allocation: RANDOMIZED
• Masking: NONE
• Purpose: BASIC SCIENCE
• Intervention Model: CROSSOVER
• Sponsor Type: OTHER
• Responsible Party: SPONSOR

Study Record Dates

• First Submitted: August 6, 2025
• First Posted: August 19, 2025
• Study Start: October 1, 2025
• Primary Completion (Estimated): September 28, 2029
• Study Completion (Estimated): December 31, 2030
• Last Updated: August 19, 2025

Record last verified: 2025-08

Data Sharing

• IPD Sharing: Will not share

Locations

PL (1)