Effects of Ozone Therapy on Muscle Damage

NCT07728422 | Completed

• 1 other identifier: 2019/188
• Study Type: interventional
• Target: 32
• Locations: 1 country
• Sites: 1

Brief Summary

The goal of this clinical trial is to determine the effectiveness of different ozone therapy methods in enhancing recovery after muscle-damaging exercise in healthy adults, and to evaluate their impact on biochemical markers of muscle damage and fatigue. The main questions it aims to answer are:

• Do ozone-based interventions accelerate recovery by reducing biochemical markers of muscle damage and fatigue following eccentric exercise?
• Which ozone therapy modality provides the greatest recovery benefit? Researchers will compare three intervention groups (autohemotherapy, ozone sauna therapy, and ozonated lavender oil massage) with a control group receiving no treatment. Participants will:
• Complete a standardized eccentric exercise protocol designed to induce muscle damage
• Receive one of the assigned interventions or no treatment (control)
• Provide blood samples at multiple time points (baseline, post-exercise, post-intervention, and 24, 48, and 72 hours follow-up) to assess recovery- related biomarkers

Conditions

Ozone Therapy; Muscle Damage; Oxidative Stress; Recovery

Keywords

Ozone theraphy; Eccentric exercise; Muscle damage biomarkers; Oxidative stress

Outcome Measures

Primary Outcomes (1)

• CK

  Serum Creatine Kinase level

  Baseline, immediately post-exercise, immediately after ozone sauna treatment, 24 hours after ozone sauna treatment, 48 hours after ozone sauna treatment and 72 hours after ozone sauna treatment.

Secondary Outcomes (4)

• LDH

  Baseline, immediately post-exercise, immediately after ozone sauna treatment, 24 hours after ozone sauna treatment, 48 hours after ozone sauna treatment and 72 hours after ozone sauna treatment.

• AST

  Baseline, immediately post-exercise, immediately after ozone sauna treatment, 24 hours after ozone sauna treatment, 48 hours after ozone sauna treatment and 72 hours after ozone sauna treatment.

• UA

  Baseline, immediately post-exercise, immediately after ozone sauna treatment, 24 hours after ozone sauna treatment, 48 hours after ozone sauna treatment and 72 hours after ozone sauna treatment.

• ALT

  Baseline, immediately post-exercise, immediately after ozone sauna treatment, 24 hours after ozone sauna treatment, 48 hours after ozone sauna treatment and 72 hours after ozone sauna treatment.

Study Arms (4)

Autohemotherapy

EXPERIMENTAL

Participants underwent major autohemotherapy involving withdrawal of venous blood, exposure to an oxygen-ozone mixture, and subsequent intravenous reinfusion in accordance with established clinical protocols

Other: Autohemotherapy

Ozone Sauna Therapy

EXPERIMENTAL

Participants underwent ozone sauna therapy administered in a sealed cabin system, with the head outside the chamber to avoid inhalation, under controlled temperature and ozone concentration conditions.

Other: Ozone Sauna Therapy

Ozonated Lavender Oil Massage

EXPERIMENTAL

Participants underwent massage therapy with ozonated lavender oil targeting lower extremity muscles, delivered by a certified therapist according to standardized procedures.

Other: Ozonated Lavender Oil Massage

Control

NO INTERVENTION

Participants remained at rest without receiving any recovery intervention, serving as the control condition for comparison.

Interventions

Autohemotherapy OTHER

Autohemotherapy was performed by withdrawing 100 mL of venous blood, which was subsequently exposed to an oxygen-ozone gas mixture (5% ozone, 95% oxygen) for 5-10 minutes in an ozone-resistant, sterile container. The ozonated blood was then rein-fused intravenously over a 15-minute period in accordance with established clinical protocols

Autohemotherapy

Ozone Sauna Therapy OTHER

Ozone sauna therapy was administered in a sealed cabin maintained at a tempera-ture of 45°C. Participants remained seated with the head positioned outside the cabin and the neck sealed using a polyethylene collar to prevent ozone inhalation. Each session lasted 20 minutes and was conducted at an ozone concentration of 20 µg·mL-¹

Ozone Sauna Therapy

Ozonated Lavender Oil Massage OTHER

Massage with ozonated lavender oil was administered by a certified massage thera-pist and targeted the lower extremity muscle groups, including the quadriceps, ham-strings, and gluteal muscles. The intervention lasted 20 minutes in total (10 minutes per limb) and followed standardized techniques adapted from Braun and Simonson (2008).

Ozonated Lavender Oil Massage

Eligibility Criteria

• Age: 18 Years - 30 Years
• Sex: male
• Healthy Volunteers: Yes
• Age Groups: Adult (18-64)

You may qualify if:

• male football players aged 18-30 years
• playing exclusively in the midfield position
• active participation in regional professional football leagues (non-top-tier)
• engagement in regular football training at least four sessions per week with a minimum duration of 90 minutes per session
• absence of acute or chronic musculoskeletal injuries within the preceding six months
• no use of anti-inflammatory medications or ergogenic supplements within the four weeks prior to the study

You may not qualify if:

• female football players
• diagnosed cardiovascular, metabolic, or neurological disorders
• known hypersensitivity to ozone exposure or lavender oil
• any surgical intervention within the past six months
• concurrent participation in alternative recovery or therapeutic interventions during the study period
• playing positions other than midfield

Sponsors & Collaborators

• Inonu University: lead

Study Sites (1)

Inonu University

Malatya, Malatya, 44280, Turkey (Türkiye)

Location

Related Publications (29)

• Dupuy O, Douzi W, Theurot D, Bosquet L, Dugue B. An Evidence-Based Approach for Choosing Post-exercise Recovery Techniques to Reduce Markers of Muscle Damage, Soreness, Fatigue, and Inflammation: A Systematic Review With Meta-Analysis. Front Physiol. 2018 Apr 26;9:403. doi: 10.3389/fphys.2018.00403. eCollection 2018.

  PMID: 29755363 BACKGROUND

• Paoli A, Bianco A, Battaglia G, Bellafiore M, Grainer A, Marcolin G, Cardoso CC, Dall'aglio R, Palma A. Sports massage with ozonised oil or non-ozonised oil: Comparative effects on recovery parameters after maximal effort in cyclists. Phys Ther Sport. 2013 Nov;14(4):240-5. doi: 10.1016/j.ptsp.2012.11.004. Epub 2013 Apr 24.

  PMID: 23623301 BACKGROUND

• Moreno-Fernandez A, Macias-Garcia L, Valverde-Moreno R, Ortiz T, Fernandez-Rodriguez A, Molini-Estrada A, De-Miguel M. Autohemotherapy with ozone as a possible effective treatment for Fibromyalgia. Acta Reumatol Port. 2019 Sep 29;44(3):244-249. Print 2019 Jul-Sep.

  PMID: 31575841 BACKGROUND

• Lin PH, Lin YP, Chen KL, Yang SY, Shih YH, Wang PY. Effect of aromatherapy on autonomic nervous system regulation with treadmill exercise-induced stress among adolescents. PLoS One. 2021 Apr 13;16(4):e0249795. doi: 10.1371/journal.pone.0249795. eCollection 2021.

  PMID: 33848307 BACKGROUND

• Pandur E, Balatinacz A, Micalizzi G, Mondello L, Horvath A, Sipos K, Horvath G. Anti-inflammatory effect of lavender (Lavandula angustifolia Mill.) essential oil prepared during different plant phenophases on THP-1 macrophages. BMC Complement Med Ther. 2021 Nov 24;21(1):287. doi: 10.1186/s12906-021-03461-5.

  PMID: 34819075 BACKGROUND

• Yousefi B, Banihashemian SZ, Feyzabadi ZK, Hasanpour S, Kokhaei P, Abdolshahi A, Emadi A, Eslami M. Potential therapeutic effect of oxygen-ozone in controlling of COVID-19 disease. Med Gas Res. 2022 Apr-Jun;12(2):33-40. doi: 10.4103/2045-9912.325989.

  PMID: 34677149 BACKGROUND

• Tartari APS, Moreira FF, Pereira MCDS, Carraro E, Cidral-Filho FJ, Salgado AI, Kerppers II. Anti-inflammatory Effect of Ozone Therapy in an Experimental Model of Rheumatoid Arthritis. Inflammation. 2020 Jun;43(3):985-993. doi: 10.1007/s10753-020-01184-2.

  PMID: 32382842 BACKGROUND

• Fatouros IG, Jamurtas AZ. Insights into the molecular etiology of exercise-induced inflammation: opportunities for optimizing performance. J Inflamm Res. 2016 Oct 21;9:175-186. doi: 10.2147/JIR.S114635. eCollection 2016.

  PMID: 27799809 BACKGROUND

• Hody S, Croisier JL, Bury T, Rogister B, Leprince P. Eccentric Muscle Contractions: Risks and Benefits. Front Physiol. 2019 May 3;10:536. doi: 10.3389/fphys.2019.00536. eCollection 2019.

  PMID: 31130877 BACKGROUND

• Powers SK, Deminice R, Ozdemir M, Yoshihara T, Bomkamp MP, Hyatt H. Exercise-induced oxidative stress: Friend or foe? J Sport Health Sci. 2020 Sep;9(5):415-425. doi: 10.1016/j.jshs.2020.04.001. Epub 2020 May 4.

  PMID: 32380253 BACKGROUND

• Davis HL, Alabed S, Chico TJA. Effect of sports massage on performance and recovery: a systematic review and meta-analysis. BMJ Open Sport Exerc Med. 2020 May 7;6(1):e000614. doi: 10.1136/bmjsem-2019-000614. eCollection 2020.

  PMID: 32426160 BACKGROUND

• Clavo B, Rodriguez-Esparragon F, Rodriguez-Abreu D, Martinez-Sanchez G, Llontop P, Aguiar-Bujanda D, Fernandez-Perez L, Santana-Rodriguez N. Modulation of Oxidative Stress by Ozone Therapy in the Prevention and Treatment of Chemotherapy-Induced Toxicity: Review and Prospects. Antioxidants (Basel). 2019 Nov 26;8(12):588. doi: 10.3390/antiox8120588.

  PMID: 31779159 BACKGROUND

• Hidalgo-Tallon FJ, Pinto-Bonilla R, Baeza-Noci J, Menendez-Cepero S, Cabizosu A. Medical ozone on hamstring injury in a professional athlete assessed by thermography: a clinical case report. BJR Case Rep. 2023 Jun 13;9(4):20220078. doi: 10.1259/bjrcr.20220078. eCollection 2023 Aug.

  PMID: 37576006 BACKGROUND

• Scassellati C, Galoforo AC, Bonvicini C, Esposito C, Ricevuti G. Ozone: a natural bioactive molecule with antioxidant property as potential new strategy in aging and in neurodegenerative disorders. Ageing Res Rev. 2020 Nov;63:101138. doi: 10.1016/j.arr.2020.101138. Epub 2020 Aug 15.

  PMID: 32810649 BACKGROUND

• Bocci VA. Scientific and medical aspects of ozone therapy. State of the art. Arch Med Res. 2006 May;37(4):425-35. doi: 10.1016/j.arcmed.2005.08.006.

  PMID: 16624639 BACKGROUND

• Ugazio E, Tullio V, Binello A, Tagliapietra S, Dosio F. Ozonated Oils as Antimicrobial Systems in Topical Applications. Their Characterization, Current Applications, and Advances in Improved Delivery Techniques. Molecules. 2020 Jan 14;25(2):334. doi: 10.3390/molecules25020334.

  PMID: 31947580 BACKGROUND

• Bocci V, Borrelli E, Valacchi G, Luzzi E. Quasi-total-body exposure to an oxygen-ozone mixture in a sauna cabin. Eur J Appl Physiol Occup Physiol. 1999 Nov-Dec;80(6):549-54. doi: 10.1007/s004210050633.

  PMID: 10541921 BACKGROUND

• Yildirim A, Belviranli M, Okudan N. Protective effect by low-intensity downhill running training against muscle damage and oxidative stress after high-intensity downhill running in rats. An Acad Bras Cienc. 2022 May 2;94(2):e20200265. doi: 10.1590/0001-3765202220200265. eCollection 2022.

  PMID: 35507975 BACKGROUND

• Kanda K, Sugama K, Sakuma J, Kawakami Y, Suzuki K. Evaluation of serum leaking enzymes and investigation into new biomarkers for exercise-induced muscle damage. Exerc Immunol Rev. 2014;20:39-54.

  PMID: 24974720 BACKGROUND

• Jamurtas AZ, Theocharis V, Tofas T, Tsiokanos A, Yfanti C, Paschalis V, Koutedakis Y, Nosaka K. Comparison between leg and arm eccentric exercises of the same relative intensity on indices of muscle damage. Eur J Appl Physiol. 2005 Oct;95(2-3):179-85. doi: 10.1007/s00421-005-1345-0. Epub 2005 Jul 9.

  PMID: 16007451 BACKGROUND

• Jang HJ, Lee JD, Jeon HS, Kim AR, Kim S, Lee HS, Kim KB. Metabolic Profiling of Eccentric Exercise-Induced Muscle Damage in Human Urine. Toxicol Res. 2018 Jul;34(3):199-210. doi: 10.5487/TR.2018.34.3.199. Epub 2018 Jul 15.

  PMID: 30057694 BACKGROUND

• Stozer A, Vodopivc P, Krizancic Bombek L. Pathophysiology of exercise-induced muscle damage and its structural, functional, metabolic, and clinical consequences. Physiol Res. 2020 Aug 31;69(4):565-598. doi: 10.33549/physiolres.934371. Epub 2020 Jul 16.

  PMID: 32672048 BACKGROUND

• Proske U, Morgan DL. Muscle damage from eccentric exercise: mechanism, mechanical signs, adaptation and clinical applications. J Physiol. 2001 Dec 1;537(Pt 2):333-45. doi: 10.1111/j.1469-7793.2001.00333.x.

  PMID: 11731568 BACKGROUND

• Allen DG. Eccentric muscle damage: mechanisms of early reduction of force. Acta Physiol Scand. 2001 Mar;171(3):311-9. doi: 10.1046/j.1365-201x.2001.00833.x.

  PMID: 11412143 BACKGROUND

• van der Horst N, Smits DW, Petersen J, Goedhart EA, Backx FJ. The preventive effect of the nordic hamstring exercise on hamstring injuries in amateur soccer players: a randomized controlled trial. Am J Sports Med. 2015 Jun;43(6):1316-23. doi: 10.1177/0363546515574057. Epub 2015 Mar 20.

  PMID: 25794868 BACKGROUND

• Van de Hoef S, Huisstede BMA, Brink MS, de Vries N, Goedhart EA, Backx FJG. The preventive effect of the bounding exercise programme on hamstring injuries in amateur soccer players: the design of a randomized controlled trial. BMC Musculoskelet Disord. 2017 Aug 22;18(1):355. doi: 10.1186/s12891-017-1716-9.

  PMID: 28830536 BACKGROUND

• Kekelekis A, Kounali Z, Kofotolis N, Clemente FM, Kellis E. Epidemiology of Injuries in Amateur Male Soccer Players: A Prospective One-Year Study. Healthcare (Basel). 2023 Jan 25;11(3):352. doi: 10.3390/healthcare11030352.

  PMID: 36766927 BACKGROUND

• Zhang Q, Dellal A, Chamari K, Igonin PH, Martin C, Hautier C. The influence of short sprint performance, acceleration, and deceleration mechanical properties on change of direction ability in soccer players-A cross-sectional study. Front Physiol. 2022 Nov 2;13:1027811. doi: 10.3389/fphys.2022.1027811. eCollection 2022.

  PMID: 36406993 BACKGROUND

• Konefal M, Chmura J, Zacharko M, Zajac T, Chmura P. The Relationship Among Acceleration, Deceleration and Changes of Direction in Repeated Small Sided Games. J Hum Kinet. 2023 Jan 4;85:96-103. doi: 10.2478/hukin-2022-0113. eCollection 2022 Dec.

  PMID: 36643839 BACKGROUND

Study Design

• Study Type: interventional
• Phase: not applicable
• Allocation: RANDOMIZED
• Masking: SINGLE
• Who Masked: PARTICIPANT
• Purpose: OTHER
• Intervention Model: PARALLEL
• Sponsor Type: OTHER
• Responsible Party: PRINCIPAL INVESTIGATOR
• PI Title: Principal Investigator

Study Record Dates

• First Submitted: July 14, 2026
• First Posted: July 27, 2026
• Study Start: October 4, 2024
• Primary Completion: November 28, 2024
• Study Completion: February 20, 2025
• Last Updated: July 27, 2026

Record last verified: 2026-07

Data Sharing

• IPD Sharing: Will share
• Shared Documents: STUDY PROTOCOL, SAP, ICF
• Time Frame: Beginning 2 months and ending 2 years after the publication of results.
• Access Criteria: Individual participant data (IPD) and supporting information will be openly available to all interested researchers. All data will be fully de-identified to ensure participant confidentiality. Researchers are encouraged to submit a brief proposal outlining the purpose and methodology of their intended analyses. Data will be shared for scientifically valid purposes, including secondary analyses, replication studies, and meta-analyses. Requests will be reviewed by the principal investigator to ensure methodological soundness and compliance with ethical standards. This study has received independent ethics committee approval, and as it involves non-sensitive data, no major access restrictions are anticipated. Data sharing will be facilitated via email request to the corresponding author. A simple data use agreement may be required. Approved users will receive de-identified datasets and relevant documentation.

Locations

TU (1)