Effect of Systemic Ozone Therapy on Corneal Endothelium

NCT07718100 | Completed

• 1 other identifier: GOKAEK/2026-01-10
• Study Type: interventional
• Target: 96
• Locations: 1 country
• Sites: 1

Brief Summary

This study evaluates the safety of systemic ozone therapy on the corneal endothelium, a vital non-renewable tissue that maintains corneal transparency. Ozone therapy is increasingly used for chronic pain, osteoarthritis, fibromyalgia, and other conditions due to its immune-modulating and antioxidant effects. However, its impact on the corneal endothelium has not been thoroughly investigated. This prospective controlled cohort study includes 96 participants: 48 patients receiving systemic ozone therapy (major autohemotherapy, 40 µg/mL, 10 sessions) and 48 age- and sex-matched healthy controls. Corneal endothelial parameters including endothelial cell density (ECD), hexagonality (Hex%), coefficient of variation (CV), central corneal thickness (CCT), and intraocular pressure (IOP) are measured by specular microscopy at baseline (T0), 1 month (T1), 3 months (T2), and 6 months (T3). Oxidative stress markers (MDA, SOD, GSH-Px) are also assessed in the ozone group. The primary goal is to determine whether systemic ozone therapy causes any adverse effects on corneal endothelial cells over a 6-month period. Secondary goals include evaluating changes in oxidative stress markers and assessing the correlation between systemic antioxidant effects and corneal health. Results will help guide clinical practice regarding the safety of ozone therapy in patients with or without pre-existing corneal conditions.

Conditions

Corneal Endothelial Cell Loss; Ozone Therapy

Keywords

Ozone Therapy; Corneal Endothelium; Specular Microscopy; Endothelial Cell Density; Oxidative Stress

Outcome Measures

Primary Outcomes (3)

• Change in Coefficient of Variation (CV) of cell area from baseline to 6 months

  Coefficient of variation in endothelial cell area (%), measured by non-contact specular microscopy (Topcon SP-3000P).

  Baseline (T0), 1 month (T1), 3 months (T2), and 6 months (T3)

• Change in Endothelial Cell Density (ECD) from baseline to 6 months

  Endothelial cell density (cells/mm²) measured by non-contact specular microscopy (Topcon SP-3000P) from the central cornea.

  Baseline (T0), 1 month (T1), 3 months (T2), and 6 months (T3)

• Change in Hexagonality (Hex%) from baseline to 6 months

  Percentage of hexagonal corneal endothelial cells measured by non-contact specular microscopy (Topcon SP-3000P).

  Baseline (T0), 1 month (T1), 3 months (T2), and 6 months (T3)

Secondary Outcomes (2)

• Change in Malondialdehyde (MDA) levels from baseline to 6 months

  Baseline (T0), 1 month (T1), 3 months (T2), and 6 months (T3)

• Change in Superoxide Dismutase (SOD) activity from baseline to 6 months

  Baseline (T0), 1 month (T1), 3 months (T2), and 6 months (T3)

Study Arms (2)

Ozone Therapy Group

EXPERIMENTAL

48 participants receiving systemic ozone therapy via major autohemotherapy (MAH) at a concentration of 40 µg/mL, administered once daily for 10 consecutive sessions.

Other: Systemic Ozone Therapy

Control Group

NO INTERVENTION

48 age- and sex-matched healthy individuals who did not receive ozone therapy.

Interventions

Systemic Ozone Therapy OTHER

Major autohemotherapy (MAH) using a medical-grade ozone generator (Ozonosan Alpha Plus, Herrmann Apparatebau GmbH, Germany). Approximately 100 mL of the patient's blood is drawn into a sterile glass bottle containing 3.8% sodium citrate as an anticoagulant. The blood is then exposed to an ozone-oxygen mixture at a concentration of 40 µg/mL. The ozonated blood is gently mixed and reinfused intravenously over 10-15 minutes. The procedure is performed once daily for 10 consecutive sessions.

Ozone Therapy Group

Eligibility Criteria

• Age: 18 Years+
• Sex: all
• Healthy Volunteers: Yes
• Age Groups: Adult (18-64), Older Adult (65+)

You may qualify if:

• Age 18 years or older
• For ozone group: Patients scheduled to receive systemic ozone therapy for fibromyalgia, chronic pain, knee osteoarthritis, disk herniation, peripheral arterial disease, or sports injuries
• For control group: Healthy individuals matched for age (±2 years), sex, and body mass index (BMI, ±3 kg/m²)
• Willing and able to provide written informed consent
• Willing and able to comply with all study procedures and follow-up visits

You may not qualify if:

• History of ocular surgery or trauma
• Corneal dystrophy or degeneration
• Glaucoma or ocular hypertension
• Diabetes mellitus
• Contact lens use within the preceding 3 months
• Active ocular surface disease
• Systemic conditions known to affect corneal endothelium
• Any condition that, in the opinion of the investigator, would interfere with study participation or evaluation of outcomes

Sponsors & Collaborators

• Yuzuncu Yil University: lead

Study Sites (1)

Yuzuncu Yil University

Van, 65080, Turkey (Türkiye)

Location

Related Publications (1)

• 1. PubMed: (Makalenin PMID'si varsa girin, yoksa boş bırakın) 2. Other: Bocci V. Ozone: A New Medical Drug. 2nd ed. Dordrecht: Springer; 2011. 3. Other: Franzini M, Valdenassi L, Pandolfi S, Tirelli U, Ricevuti G, Chirumbolo S. The role of ozone as an Nrf2-Keap1-ARE activator in the anti-microbial activity and immunity modulation of infected wounds. Antioxidants. 2023;12(11):1985. doi:10.3390/antiox12111985 4. Other: Malatesta M, Tabaracci G, Pellicciari C. Low-dose ozone as a eustress inducer: Experimental evidence of the molecular mechanisms accounting for its therapeutic action. Int J Mol Sci. 2024;25(23):12657. doi:10.3390/ijms252312657 5. Other: Cheng Y, Lu Y, Du Y, Zhao Y, Zhuang X. A clinical study on ozone autohemotherapy for the treatment of acute ischemic stroke. Front Neurol. 2025;16:1583726. doi:10.3389/fneur.2025.1583726

  BACKGROUND

Related Links

• Study dataset and supporting information on Zenodo

MeSH Terms

Conditions

Corneal Endothelial Cell Loss

Condition Hierarchy (Ancestors)

Corneal Diseases; Eye Diseases; Eye Manifestations; Postoperative Complications; Pathologic Processes; Pathological Conditions, Signs and Symptoms; Signs and Symptoms

Study Design

• Study Type: interventional
• Phase: not applicable
• Allocation: NON RANDOMIZED
• Masking: NONE
• Masking Details: Specular microscopy measurements were performed by a single experienced technician who was blinded to group allocation. However, the study was open-label for participants and investigators.
• Purpose: OTHER
• Intervention Model: PARALLEL
• Sponsor Type: OTHER
• Responsible Party: PRINCIPAL INVESTIGATOR
• PI Title: Professor

Model Details: Participants were assigned to either the ozone therapy group (major autohemotherapy, 40 µg/mL, 10 sessions) or the control group (no intervention). Both groups were followed prospectively over 6 months with serial measurements at baseline, 1 month, 3 months, and 6 months.

Study Record Dates

• First Submitted: July 16, 2026
• First Posted: July 21, 2026
• Study Start: January 14, 2026
• Primary Completion: July 14, 2026
• Study Completion: July 14, 2026
• Last Updated: July 21, 2026

Record last verified: 2026-07

Data Sharing

• IPD Sharing: Will share
• Time Frame: Data will be available starting 6 months after publication and ending 3 years after publication.
• Access Criteria: Data access will be granted upon reasonable request to the corresponding author after approval of a research proposal.

Locations

TU (1)