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 Table of Contents  
Year : 2017  |  Volume : 7  |  Issue : 3  |  Page : 212-219

Ozone therapy: An overview of pharmacodynamics, current research, and clinical utility

1 Foley Plaza Medical, New York, NY, USA
2 Department of Physiology and Biophysics, Stony Brook University School of Medicine, Stony Brook, NY, USA
3 Department of Physiology and Biophysics, Stony Brook University School of Medicine, Stony Brook, NY, USA; Medical Student Research Institute, St. George's University School of Medicine, Grenada, West Indies
4 Department of Cardiothoracic Surgery, Lenox Hill Hospital, New York, NY, USA
5 Department of Physiology and Biophysics, Stony Brook University School of Medicine, Stony Brook, NY; Department of Urology, Stony Brook University School of Medicine, Stony Brook, NY, USA

Date of Web Publication17-Oct-2017

Correspondence Address:
Sardar Ali Khan
Department of Physiology and Biophysics, Stony Brook University School of Medicine, Stony Brook, NY; Department of Urology, Stony Brook University School of Medicine, Stony Brook, NY
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Source of Support: None, Conflict of Interest: None

DOI: 10.4103/2045-9912.215752

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The use of ozone (O3) gas as a therapy in alternative medicine has attracted skepticism due to its unstable molecular structure. However, copious volumes of research have provided evidence that O3's dynamic resonance structures facilitate physiological interactions useful in treating a myriad of pathologies. Specifically, O3 therapy induces moderate oxidative stress when interacting with lipids. This interaction increases endogenous production of antioxidants, local perfusion, and oxygen delivery, as well as enhances immune responses. We have conducted a comprehensive review of O3 therapy, investigating its contraindications, routes and concentrations of administration, mechanisms of action, disinfectant properties in various microorganisms, and its medicinal use in different pathologies. We explore the therapeutic value of O3 in pathologies of the cardiovascular system, gastrointestinal tract, genitourinary system, central nervous system, head and neck, musculoskeletal, subcutaneous tissue, and peripheral vascular disease. Despite compelling evidence, further studies are essential to mark it as a viable and quintessential treatment option in medicine.

Keywords: ozone; ozone therapy; ozone gas; autohemotherapy; oxidative stress; reactive oxidative species; lipid ozonation products; oxidative preconditioning

How to cite this article:
Smith NL, Wilson AL, Gandhi J, Vatsia S, Khan SA. Ozone therapy: An overview of pharmacodynamics, current research, and clinical utility. Med Gas Res 2017;7:212-9

How to cite this URL:
Smith NL, Wilson AL, Gandhi J, Vatsia S, Khan SA. Ozone therapy: An overview of pharmacodynamics, current research, and clinical utility. Med Gas Res [serial online] 2017 [cited 2023 Mar 31];7:212-9. Available from: https://www.medgasres.com/text.asp?2017/7/3/212/215752

  Introduction Top

Ozone (O3) gas was discovered in the 1840s, and soon after that, the scientific community began to expand past the notion that it was just another gas of the Earth's atmosphere. Though the migration of O3 into the medical field has taken a circuitous road since the 19th century, its medicinal value is currently controversial despite compelling research.[1] O3 is highly water-soluble inorganic molecule composed of three oxygen molecules. O3's inherently unstable molecular structure, due to the nature of its mesomeric states, tends to make it difficult to obtain high concentrations. O3 will often experience transient reactions with itself or water. Thus, it was initially problematic to achieve desired levels and even more difficult is to assess the therapeutic effects of such a transient state.[1],[2] These mesomeric states create a conundrum within the scientific community. A divide has formed between those who believe the volatile nature of these mesomeric states can foster positive responses and those who are wary of its seemingly dangerous effects.

Despite suspicions, a multitude of O3 therapies have shown substantial benefits that span a large variety of acute and chronic ailments. O3 is currently prevalent in dentistry to treat diseases of the jaw.[1] O3 has also proven itself beneficial as a disinfectant for drinking water and sterilization of medical instruments.[1],[3] The function of O3 shares similarities to that of a prodrug, as it is modified upon reacting with molecules to create more active substrates, thus stimulating an endogenous cascade of responses. On the other hand, it is hard to classify O3 as simply a prodrug, due to its capability to directly interact with phospholipids, lipoproteins, cell envelopes of bacteria, and viral capsids. The physiology of these biological responses is herein discussed.

Despite the various benefits, O3 toxicity and clinical utility depends on the concentration and administration to the appropriate site.[1],[2],[4],[5] One of the major contraindications of O3 therapy is lung inhalation. O3 therapy significantly increases airway resistance without changing the compliance or elastic characteristics of the lung.[1] Additionally, direct contact of O3 with the eyes and lungs is contraindicated because of the low antioxidant capabilities in these specific locations.[6]

  Literature Retrieval Top

A MEDLINE® database search of literature extended from 1980 to 2017 to obtain current information regarding O3 therapy, its routes of administration, and mechanism of action. Subsequently, trials pertaining to the clinical implications of O3 therapy were paired by pathology and anatomical system. The most important points refer to the type of pathology, route of O3 administration, type of research trial, result(s) of the trial, side effect(s), and proposed physiological mechanism(s). Literature retrieval was performed in July 2017 and included the term “ozone therapy” combined with the following search criteria: “routes of administration”, “mechanism of action”, “cardiovascular”, “subcutaneous tissue”, “peripheral vascular disease”, “neurological”, “head and neck”, “orthopedic”, “musculoskeletal”, “gastrointestinal”, and “genitourinary”. We did not formulate any exclusion criteria.

  Routes of Administration Top

O3 therapy combines a mixture of oxygen (O2)-O3, with a diverse therapeutic range (10–80 μg/ml of gas per ml of blood).[5],[6],[7] O3 therapy administration is variable based on treatment goals and location of therapy. The first and most popular is O3 autohemotransfusion (O3-Aht). O3-Aht has grown in popularity because it allows for a predetermined amount of blood to be taken and thus, using stoichiometric calculations, a precise concertation of O2-O3 can be infused. This small amount of blood is subjected to O2-O3 ex vivo is then administered to the patient.[5],[6] Extracorporeal blood oxygenation and ozonation are very similar techniques. However, its goal is to obtain higher blood volume than the 200–300 mL seen in O3-Aht.[5]

Other modalities of therapies include direct injection Via the intramuscular, intradiscal, and paravertebral site of administration. Rectal insufflation of O3-O3 is another common site of administration. However, insufflation of the nasal, tubal, oral, vaginal, vesical, pleural, and peritoneal cavities have proven to be prudent routes of administration. Cutaneous exposure has also had likely outcomes and can be achieved by sealing the portion of the body in a chamber or bag and insufflating with O3-O3 mixture. Saline with O3-O3 dissolved is used to avoid the risk of embolism when administered intravenously.[5]

  Mechanism of Action Top

Antioxidant capacity

Upon beginning O3 therapy, a multifaceted endogenous cascade is initiated and releases biologically active substrates in response to the transient, and moderate, oxidative stress that O3 induces. O3 can cause this mild oxidative stress because of its ability to dissolve in the aqueous component of plasma.[8] By reacting with polyunsaturated fatty acids (PUFA) and water, O3 creates hydrogen peroxide (H2O2), a reactive oxygen species (ROS). Simultaneously, O3 forms a mixture of lipid ozonation products (LOP).[9] The LOPs created after O3 exposure include lipoperoxyl radicals, hydroperoxides, malonyldialdeyde, isoprostanes, the ozonide and alkenals, and 4-hydroxynonenal (4-HNE).

Moderate oxidative stress caused by O3 increases activation of the transcriptional factor mediating nuclear factor-erythroid 2-related factor 2 (Nrf2). Nrf2's domain is responsible for activating the transcription of antioxidant response elements (ARE). Upon induction of ARE transcription, an assortment of antioxidant enzymes gains increased concentration levels in response to the transient oxidative stress of O3. The antioxidants created include, but are not limited to, superoxide dismutase (SOD), glutathione peroxidase (GPx), glutathione S-transferase (GST), catalase (CAT), heme oxygenase-1 (HO-1), NADPH-quinone-oxidoreductase (NQO-1), heat shock proteins (HSP), and phase II enzymes of drug metabolism. Many of these enzymes act as free radical scavengers clinically relevant to a wide variety of diseases.[9]

O3, as well as other medical gases, e.g., carbon monoxide (CO) and nitric oxide (NO), has twofold effects depending on the amount given and the cell's redox status. There is a complex relationship between these three medical gases as O3 overexpresses HO-1, also referred to as HSPs of 32 kPa (Hsp32),[10] the enzyme responsible for CO formation, and downregulates NO synthase, which generates NO. Furthermore, O3 upregulates the expression levels of Hsp70 which, in turn, is strictly related to HO-1. O3 may have a developing role in Hsp-based diagnosis and therapy of free radical-based diseases. HO-1 degrades heme, which can be toxic depending on the amount produced, into free iron, CO, and biliverdin (i.e., precursor of bilirubin), a neutralizer of oxidative and nitrosative stress due to its ability to interact with NO and reactive nitrogen species.[11],[12] Recently, it is becoming clear the heat shock response (HSR) provides a cytoprotective state during inflammation, cancer, aging, and neurodegenerative disorders.[13] Given its extensive cytoprotective properties, the HSR is now a target for induction via pharmacological agents.[1] Hsp70 is involved in co- and post-translational folding, the quality control of misfolded proteins,[14] folding and assembly of de novo proteins into macromolecular complexes, as well as anti-aggregation, protein refolding, and degradation.[15] HO isoforms are acknowledged as dynamic sensors of cellular oxidative stress and regulators of redox homeostasis throughout the phylogenetic spectrum. The effect of O3 on these cell activities remains to be evaluated. Hormesis is a potent, endogenous defense mechanism for lethal ischemic and oxidative insults to multiple organ systems.[13] O3 may have a hormetic role in regulating the anti-inflammatory and proinflammatory effects of CO, including prostaglandin formation akin to NO, which has been shown to exert some of its biological actions through the modulation of prostaglandin endoperoxide synthase activity.[16] Inhibiting HO activity prevents CO biosynthesis and its downstream effects[17]; the effect of O3 on this cascade is yet to be determined.

Animal models have postulated the beneficial effects of prophylactic O3 therapy in controlling the age-related effects of oxidative stress.[18],[19] Evidence was provided to show that low O3 dose administration provided beneficial effects on age-related alterations in the heart and hippocampus of rats. Additional research has been performed and provided room for speculation that O3 therapy may provide the mediation of a mechanism involved in rebalancing the dysregulated redox state that accumulates as individuals age.[20] There was an apparent reduction of lipid and protein oxidation markers, lessening of lipofuscin deposition, restoration of glutathione (GSH) levels, and normalization of GPx activity in aged heart tissue. O3 was demonstrated to decrease age-associated energy failure in the heart and hippocampus of rats. Researchers suspect that the improved cardiac cytosolic calcium and restoration of weakened Na+-K+ ATPase activity in the heart and hippocampus, respectively, were associated with the improvements seen.[20]

In hopes of attaining a sense of the possible toxic components of O3 therapy, a study was done to assess the extent of lesions on human hematic mononucleated cells (HHMC), human thymic epithelium, murine macrophages, mouse splenocytes, and B16 melanoma murine cells. A significant finding of the study was that Hsp70 exhibited an O3-induced increase in biosynthesis in HHMC. Hsp70s are synthesized in response to thermal shock and other stressing agents to cope with the damage that stimulates their biosynthesis.[21] Additionally, they stimulate several immune system responses in lymphocytes and macrophages. The study provided evidence that O3 is a stressing agent capable of upregulating the biosynthesis of Hsp70, without toxicity to membranes. However, the membranes of macrophages are highly resistant to the possible toxicity of O3 at high concentrations; HHMC is less resistant at the high end of the spectrum. The statement above should not discount the effectiveness of O3 as a therapy because Hsp70s are induced in HHMCs without lesions up to 20 μg/mL— a typical dose given in O3-AHT.[21]

Cisplatin (CDDP), a treatment used in a variety of cancers has been observed to have nephrotoxicity in 25% of the patients as a side effect. The occurrence of this nephrotoxicity is thought to be secondary to the free radical generation and the inability of ROS scavengers to ameliorate these molecules, leading to acute renal failure. O2-O3 therapy was used to increase the antioxidant capacity of rats exposed to CDDP and compared to control groups. Serum creatine levels were significantly reduced compared to control groups, illustrating the decreased nephrotoxicity indirectly in the rats with CDDP and O2-O3 therapy. In addition to attenuating the nephrotoxicity, O2-O3 therapy also restores the levels of antioxidant defense constituents (GSH, SOD, CAT, and GSH-Px), which are usually decreased by CDDP. Also, thiobarbituric acid reactive substances (TBARS) were reduced, which is a marker of lipid peroxidation in the kidney.[22],[23]

Additional human studies examined the beneficial effects of O3 therapy employed via O3-AHT, in conjunction with coenzyme Q10, administered orally. The study evaluated SOD levels, a powerful antioxidant and catalase enzyme, an additional antioxidant enzyme in a control group, a group of O3 therapy by itself, and O3 therapy combined with Q10. Evidence has implied that SOD was significantly increased and catalase enzyme insignificantly increased in the O3 + Q10 group when compared to the control group. Malondialdehyde, a product of lipid peroxidation, is an indicator of oxidative membrane damage. Malondialdehyde levels were significantly decreased concentrations in the O3 + Q10 group when compared to the control group. Taken together, this study provides evidence of the beneficial effects of O3 therapy in combination with Q10 in combatting and the prevention of damage elicited by oxidation.[9]

Multiple studies have provided evidence that O3 therapy increased activation of the Nrf2 pathway via the induction of moderate oxidative stress.[15],[24] By doing so, a transient increase in H2O2 and LOPs enhances the number of antioxidants and therefore can be used for a longer time frame to re-establish the balance of the redox system. Additionally, the creation of these antioxidant enzymes has effects, not only at the level of O3 radical metabolism, but on the whole body.[22],[23]

Researchers have argued that knowing the total antioxidant status and plasma protein thiol group levels of a blood sample are indicators of the precise amount of O3 required to optimize treatments. By developing more accurate antioxidant status indicators, an individual treatment would achieve the correct dosage on a day and case basis.[7],[23],[25] Systems have been proposed to have a more precise measurement of the redox state of a patient to achieve this goal. One system proposes simultaneously measuring different biological markers in the blood such as GSH, GPx, GST, SOD, CAT, conjugated dienes, total hydroperoxides, and TBARS. Using an algorithm, information can be gathered about the total antioxidant activity, total pro-oxidant activity, redox index, and grade of oxidative stress. Systems like this can provide insights to the correct dosage and response to O3 therapy based on oxidative stress levels seen in the patient.[7],[23],[25]

Vascular and hematological modulation

O3 is a stimulator of the transmembrane flow of O2. The increase in O2 levels inside the cell secondary to O3 therapy makes the mitochondrial respiratory chain more efficient.[26] In red blood cells, O3-AHT may increase the activity of phosphofructokinase, increasing the rate of glycolysis. By enhancing the glycolytic rate, there is an increase in ATP and 2,3-diphosphoglycerate (2,3-DPG) in the cell. Subsequently, due to the Bohr effect, there is a rightward shift in the oxyhemoglobin dissociation curve allowing for the oxygen bound hemoglobin to be unloaded more readily to ischemic tissues. Combined with the increase in NO synthase activity, there is a marked increase in perfusion to the area under stimulation by O3-AHT.[27] With repeated treatment, sufficient enough LOP may be generated to reach the bone marrow acting as repeated stressors to simulate erythrogenesis and the upregulation of antioxidant enzyme upregulation. O3 also causes a reduction in nicotinamide adenine dinucleotide (NADH) and assists in the oxidation of cytochrome c.[1],[28]

O3 has also been shown to improve blood circulation and oxygen delivery to ischemic tissues.[29] Multiple studies have provided evidence that the correction of chronic oxidative stress via the increase of antioxidant enzymes in O3 can increase erythroblast differentiation. This leads to a progressive increase in erythrocytes and preconditions them to having resilience towards oxidative stress. This is known as “oxidative preconditioning”.[1],[30] Also, O3 increases levels of prostacyclin, a known vasodilator.[1]

Additionally, it was speculated that O3's oxidative capabilities would interfere with the endothelial production of NO and thus hinder vasodilation. However, studies have provided evidence that because NO is not substantially transported in the vasculature of the blood, a deleterious interaction is unlikely.[29] Since HO-derived bilirubin31 has been demonstrated to interact with NO,[11],[12] O3-induced HO upregulation could modify NO production and alter vasodilation.

Unpredictably, studies have shown an increase of NO, which led to speculation of O3's ability to activate genes associated with NO synthase expression to further promote higher levels of NO formation. Moreover, O3's stimulation of antioxidant enzymes are also speculated to increase NO levels. While endothelial generation of superoxide disrupts the activity of NO, O3 upregulates the enzymes to ameliorate the downstream effects of ROS responsible for deleterious vasoconstriction.[29],[32]

The prophylactic role of O3 has been explored with hepatic ischemia/reperfusion (I/R) injury, a phenomenon associated with liver transplantation. Hepatic I/R is a clinically unsolved problem mainly due to the unknown mechanisms that are the foundations of this ailment. In summary, O3 oxidative preconditionings (ozoneOPs) were found to protect against liver I/R injury through mechanisms that promote a regulation of endogenous NO concentrations and the maintenance of an adequate cellular redox balance. OzoneOPs are postulated to upregulate endogenous antioxidant systems and generate an increase in NO molecule generation, both of which are protective orders against liver and pancreas damage. The results in this animal model provided evidence that ozoneOPs protected against liver I/R via an increase in concentrations of endogenous NO and prime cells to have a more balanced redox system.[32] Additionally, enhanced activation of adenosine A1 receptors in rat models have been observed with ozoneOPs in liver I/R.[33]

Further studies have expanded upon this postulation by applying O3 therapy to renal I/R in rats. Renal I/R is a primary cause of acute renal failure after transplantation surgery. The findings of a study by Orakdogen et al. [34] indicated that the ozoneOPs allowed for a protective element when facing I/R. Following an increase in endothelial NO synthase and inducible NO synthase expression, it was concluded that ozoneOPs were intimately related to the increasing NO production as well as reducing renal damage by suppressing endothelin 1.[34]

Cerebral vasospasm after subarachnoid hemorrhage is a significant detriment to the recovery of patients. An animal model examined the effects intravenous O3 therapy on vasospasms in the rat femoral artery. Histopathological and morphometric measurements provided evidence that O3 therapy decreased morphometric changes, disruption of endothelial cells, and hemorrhages that are a result of vasospasm. The study speculated the anti-oxidative and anti-inflammatory effects of O3 might be a prudent treatment for posthemorrhagic vasospasm.[35]

Pathogen inactivation

When bacteria are exposed to O3 in vitro, the phospholipids, and lipoproteins that are within the bacterial cell envelope are oxidized. As this occurs, the stability of the bacterial cell envelope is attenuated. Moreover, evidence has demonstrated O3 to interact with fungal cell walls like bacteria. This disrupts the integrity of the cytosolic membrane and infiltrates the microorganisms to oxidize glycoproteins, glycolipids, and block enzymatic function. The combination of these reactions causes inhibition of fungi growth and mortality of bacteria and fungi.[1],[3],[5] In vitro, O3 has been shown to interfere with virus-to-cell contact in lipid-enveloped viruses via oxidation of lipoproteins, proteins, and glycoproteins, thus interfering with the viral reproductive cycles.[1],[3],[36]

Specifically, animal models have shown that O3 therapy as an adjunct to vancomycin enhances the animal's capability to eliminate methicillin-resistant Staphylococcus aureus mediastinitis.[37]

Immune system activation

In vivo, O3 therapy has been shown to have multifaceted effects when interacting with PUFA. As stated previously, O3 reacts with PUFA and other antioxidants, H2O2 and varies peroxidation compounds are formed. H2O2 readily diffuses into immune cells has been shown to act as a regulatory step in signal transduction and facilitating a myriad of immune responses.[36],[38] Specifically, increases in interferon, tumor necrosis factor, and interleukin (IL)-2 are seen. The increases with IL-2 are known to initiate immune response mechanisms.[1] Additionally, H2O2 activates nuclear factor-kappa B (NF-κB) and transforming growth factor beta (TGF-β), which increase immunoactive cytokine release and upregulate tissue remodeling. H2O2 mediates the action of NF-κB by enhancing the activity of tyrosine kinases that will phosphorylate IκB, a subunit of the transcription factor NF-κB.[34],[37] Low doses of O3 have been shown to inhibit prostaglandin synthesis, release bradykinin, and increase secretions of macrophages and leukocytes.[34] Having the correct amount of either of these oxidative markers can be used to create a sufficient rise in H2O2 and NO levels to stimulate the most notable increase in IL-8. IL-8 also activates NF-κB, allowing production of ROS scavengers.[7]

Animal models using O3 have shown to reduce and prevent inflammatory responses steming from the presence of E. coli in the renal system.[26],[38] Additional studies have provided evidence of the anti-inflammatory effects of O3. A study by Chang et al. [25] purified rheumatoid arthritis synovial fibroblast cells from human patients and injected them into immunocompromised mouse joints. Using an Ozonsan-α generator to deliver precise gas flows to vessels in the localized area, the authors discovered that 3% and 5% O3 application significantly decreased the proinflammatory cytokines IL-1β, IL-6, and TNF-α without any toxicity or severe side effects.[25]

Studies have shown that human cancer cells from lung, breast, and uterine tumors are inhibited in a dose-dependent manner by O3 therapy in vitro. O3 concentrations of 0.3 and 0.5 ppm inhibited cancer cell growth by 40% and 60%, respectively. Furthermore, the noncancerous cell controls were not affected by these levels of O3. At 0.8 ppm, cancer cell growth was inhibited by more than 90%. However, the control cell growth was less than 50%. Additionally, as control cells aged, they exhibited further growth inhibition and morphological changes. The study speculated that as the healthy cells matured, there was a decrease in growth due to the increased cellular damage incurred by each division.[39]

  Clinical Utility Top

With its ever-growing ubiquity, O3 therapy is finding a place in many branches of medicine and medical specialties. In fact, its clinical use can be arranged systematically into cardiovascular (Additional Table 1 [Additional file 1]), subcutaneous tissue (Additional Table 2 [Additional file 2]), peripheral vascular disease (Additional Table 3 [Additional file 3]), neurological (Additional Table 4 [Additional file 4]), head and neck (Additional Table 5 [Additional file 5]), orthopedic (Additional Table 6 [Additional file 6]), gastrointestinal (Additional Table 7 [Additional file 7]), and genitourinary (Additional Table 8 [Additional file 8]). These indications are a product of human clinical trials conducted for specific pathologies related to the aforementioned systems. Despite a lack of direct support of O3 therapy, the current Food and Drug Administration regulations do not restrict the use of it in situ ations where it has proven its safety and effectiveness. Nonetheless, there has been support for its safety and effectiveness in multi-international studies.

  Conclusions Top

O3 therapy can alter the natural history of several disease and disorders, with potentially many more yet untested. A plethora of laboratory studies have provided evidence of O3's antioxidant capabilities, as well as vascular, hematological, and immune system modulations. This evidence has been further substantiated in clinical trials with O3 therapy being useful in the cardiovascular, subcutaneous tissue, peripheral vascular disease, neurological, head and neck, orthopedic, gastrointestinal, and genitourinary pathologies. O3 therapy has proven especially beneficial in the diabetic foot, ischemic wounds, and peripheral vascular disease, areas in which O3 use is most prevalent. Upcoming laboratory and translational research should begin to develop protocols for O3-AHT in attempts to establish a dose-response relationship as it has demonstrated high utility in a myriad of pathologies at varying concentrations. Despite the presently compelling evidence, future studies should include more double-blind, randomized clinical trials with greater sample sizes, determination of longevity in benefits produced, as well as methods of measurements and analysis.


The authors are thankful to Drs. Kelly Warren, Inefta Reid, Todd Miller, and Peter Brink (Department of Physiology and Biophysics, Stony Brook University School of Medicine, Stony Brook, NY, USA) for departmental support, as well as Mrs. Wendy Isser and Ms. Grace Garey (Northport VA Medical Center Library, Northport, NY, USA) for literature retrieval.

Author contributions

NLS designed, organized, and wrote the article; ALW designed the outline, wrote the article, and solved queries related to scientific publications from the journals; JG performed literature searches, critiqued the literature findings, and wrote the article; SV critiqued and applied logical reasoning to the literature findings; SAK applied clinical concepts, revised the article to add logical reasoning, and cross-checked the referencing. All authors have read and approved the manuscript provided.

Conflicts of interest

The authors have no conflicts of interest to declare.

Plagiarism check

Checked twice by iThenticate.

Peer review

Externally peer reviewed.

Open peer reviewers

Ozan Akca, University of Louisville, USA; Nemoto Edwin, University of New Mexico Health Sciences Center, USA; Mancuso Cesare, Università Cattolica del Sacro Cuore, Italy.

Additional files

Additional Table 1: Cardiovascular indications for O3 therapy.

Additional Table 2: Subcutaneous tissue indications for O3 therapy.

Additional Table 3: Peripheral vascular disease indications for O3 therapy.

Additional Table 4: Neurological indications for O3 therapy.

Additional Table 5: Head and neck indications for O3 therapy.

Additional Table 6: Orthopedic indications for O3 therapy.

Additional Table 7: Gastrointestinal indications for O3 therapy.

Additional Table 8: Genitourinary indications for O3 therapy.[78]

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53 Topical Liposomal Ozonated Oil in Complicated Corneal Disease: A Report on Three Clinical Cases
Fedele Passidomo,Francesco Pignatelli,Giuseppe Addabbo,Ciro Costagliola
International Medical Case Reports Journal. 2021; Volume 14: 327
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54 The interference of ozone gas in kinects and mitochondrial potential of equine sperm submitted on cryopreservation
Iara Nóbrega Macêdo, Lucia Cristina Pereira Arruda, Breno Barros de Santana, Thalles Cloves Maciel de Moura, Maria Madalena Pessoa Guerra, Diogo Gutemberg Bezerra, Gustavo Ferrer Carneiro, Sildivane Valcácia Silva
Animal Reproduction. 2021; 18(4)
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55 Does paravertebral ozone injection have efficacy as an additional treatment for acute lumbar disc herniation? A randomized, double-blind, placebo-controlled study
Hamza Sucuoglu,Nalan Soydas
Journal of Back and Musculoskeletal Rehabilitation. 2021; : 1
[Pubmed] | [DOI]
56 The Relationship between Ozone and Human Blood in the Course of a Well-Controlled, Mild, and Transitory Oxidative Eustress
Gerardo Tricarico, Valter Travagli
Antioxidants. 2021; 10(12): 1946
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57 Topical Application of Ozonated Oils for the Treatment of MRSA Skin Infection in an Animal Model of Infected Ulcer
Vanessa Silva,Cecília Peirone,Rosa Capita,Carlos Alonso-Calleja,José A. Marques-Magallanes,Isabel Pires,Luís Maltez,José Eduardo Pereira,Gilberto Igrejas,Patrícia Poeta
Biology. 2021; 10(5): 372
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58 Ozone Gel in Chronic Periodontal Disease: A Randomized Clinical Trial on the Anti-Inflammatory Effects of Ozone Application
Marco Colombo,Simone Gallo,Alessandro Garofoli,Claudio Poggio,Carla Renata Arciola,Andrea Scribante
Biology. 2021; 10(7): 625
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59 Oxygen–Ozone Therapy in the Rehabilitation Field:State of the Art on Mechanisms of Action, Safety andEffectiveness in Patients with Musculoskeletal Disorders
Alessandro de Sire,Francesco Agostini,Lorenzo Lippi,Massimiliano Mangone,Simone Marchese,Carlo Cisari,Andrea Bernetti,Marco Invernizzi
Biomolecules. 2021; 11(3): 356
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60 Ozone as Modulator of Resorption and Inflammatory Response in Extruded Nucleus Pulposus Herniation. Revising Concepts
María de los Ángeles Erario,Eduardo Croce,Maria Teresita Moviglia Brandolino,Gustavo Moviglia,Aníbal M. Grangeat
International Journal of Molecular Sciences. 2021; 22(18): 9946
[Pubmed] | [DOI]
61 Cold helium plasma as a modifier of free radical processes in the blood: in vitro study
Andrew K. Martusevich,Alexander G. Galka,Konstantin A. Karuzin,Alexander N. Tuzhilkin,Svetlana L. Malinovskaya
AIMS Biophysics. 2021; 8(1): 34
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62 Effect of topical ozonated sunflower oil on second intention wound healing in turtles: a randomised experimental study
Pedro J. Ginel,Joao Negrini,Rafael Guerra,Rosario Lucena,María T. Ruiz-Campillo,Elena Mozos
Journal of Veterinary Science. 2021; 22
[Pubmed] | [DOI]
63 Ozonized Gel Against Four Candida Species: A Pilot Study and Clinical Perspectives
Vincenzina Monzillo,Fabiola Lallitto,Alba Russo,Claudio Poggio,Andrea Scribante,Carla Renata Arciola,Francesco Rocco Bertuccio,Marco Colombo
Materials. 2020; 13(7): 1731
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64 Anticancer Effects of Cold Atmospheric Plasma in Canine Osteosarcoma Cells
Jaehak Lee,Hyunjin Moon,Bonghye Ku,Keunho Lee,Cheol-Yong Hwang,Seung Joon Baek
International Journal of Molecular Sciences. 2020; 21(12): 4556
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65 Recent Developments in Ozone Sensor Technology for Medical Applications
Lisa Petani,Liane Koker,Janina Herrmann,Veit Hagenmeyer,Ulrich Gengenbach,Christian Pylatiuk
Micromachines. 2020; 11(6): 624
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66 Ozonated Oils as Antimicrobial Systems in Topical Applications. Their Characterization, Current Applications, and Advances in Improved Delivery Techniques
Elena Ugazio,Vivian Tullio,Arianna Binello,Silvia Tagliapietra,Franco Dosio
Molecules. 2020; 25(2): 334
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67 High Efficacy of Ozonated Oils on the Removal of Biofilms Produced by Methicillin-Resistant Staphylococcus aureus (MRSA) from Infected Diabetic Foot Ulcers
Vanessa Silva,Cecília Peirone,Joana S. Amaral,Rosa Capita,Carlos Alonso-Calleja,José A. Marques-Magallanes,Ângela Martins,Águeda Carvalho,Luís Maltez,José Eduardo Pereira,José Luís Capelo,Gilberto Igrejas,Patrícia Poeta
Molecules. 2020; 25(16): 3601
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68 Mechanisms and practical use of the bactericidal effects of ozone and ozonated oils
S. Schetinin
Terapevt (General Physician). 2020; (6): 45
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69 La importancia de reducir la carga viral para disminuir el riesgo de contagio por COVID-19
 Livia  Contreras-Bulnes,David Rodríguez-Marín, Rufino  Iribarren-Moreno,Javier Bucio-Duarte, Carlos de J  Álvarez-Díaz,Estela Vélez-Benítez, Guillermo  Ramos-Gallardo,Jesús Cuenca-Pardo
Cirugía Plástica. 2020; 30(2): 78
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70 The importance of reducing the viral load to diminish the risk of COVID-19 spread
 Livia  Contreras-Bulnes,David Rodríguez-Marín, Rufino  Iribarren-Moreno,Javier Bucio-Duarte, Carlos de J  Álvarez-Díaz,Estela Vélez-Benítez, Guillermo  Ramos-Gallardo,Jesús Cuenca-Pardo
Cirugía Plástica. 2020; 30(2): 78
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71 Ozone Exposure Induces Metabolic Disorders and NAD+ Depletion Through PARP1 Activation in Spinal Cord Neurons
Shulin Ma,Xu Zhao,Cong Zhang,Panpan Sun,Yun Li,Xiaowen Lin,Tao Sun,Zhijian Fu
Frontiers in Medicine. 2020; 7
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72 The effects of ozone therapy on postoperative adhesions and ovarian functions: An experimental study
Rulin DENIZ,Yakup BAYKUS,Muhammet Bora UZUNER,Yasemen ADALI
Journal of Surgery and Medicine. 2020;
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73 The effect of ozone injection within a common peroneal nerve schwannoma: A mistreatment due to a misdiagnosis
Ignazio Gaspare Vetrano,Francesco Acerbi,Gianluca Marucci,Vittoria Nazzi
Surgical Neurology International. 2020; 11: 413
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74 Ozone therapy with local cellular immune modulation and disc progenitor cell implant is safe, effective and efficient
AM Grangeat, EA Crocce, MA Erario, MT Moviglia Brandolino, SL Piccone, MA Lopez, GA Moviglia
Open Journal of Orthopedics and Rheumatology. 2020; 5(1): 024
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75 Modification of Hypoxic States in Photodynamic Therapy
A. B. Gapeyev,T. G. Shcherbatyuk
Biochemistry (Moscow), Supplement Series A: Membrane and Cell Biology. 2020; 14(3): 184
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76 Antimycotic Activity of Ozonized Oil in Liposome Eye Drops against Candida spp.
Giuseppe Celenza,Roberto Iorio,Salvatore Cracchiolo,Sabrina Petricca,Ciro Costagliola,Benedetta Cinque,Bernardetta Segatore,Gianfranco Amicosante,Pierangelo Bellio
Translational Vision Science & Technology. 2020; 9(8): 4
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77 Comment on “Unknown Challenge #8: Pneumomastia”: Additional Differential Diagnosis to Pneumomastia
Haydee Ojeda-Fournier
Journal of Breast Imaging. 2020;
[Pubmed] | [DOI]
78 AMPK activation by ozone therapy inhibits tissue factor-triggered intestinal ischemia and ameliorates chemotherapeutic enteritis
Qingqing Yu,Xing Yang,Chen Zhang,Xiaotao Zhang,Chaoyu Wang,Lu Chen,Xiaolin Liu,Yufeng Gu,Xueming He,Liang Hu,Wen-Tao Liu,Yan Li
The FASEB Journal. 2020;
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79 Effects of ozone for treating chronically refractory wounds and ulcers
Qing Wen,Dongying Liu,Xian Wang,Yanli Zhang,Song Fang,Xianliang Qiu,Qiu Chen
Medicine. 2020; 99(22): e20457
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80 Ozone therapy in veterinary medicine: A review
R.L. Sciorsci,E. Lillo,L. Occhiogrosso,A. Rizzo
Research in Veterinary Science. 2020; 130: 240
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81 Effects of ozone treatment on penile erection capacity and nitric oxide synthase levels in diabetic rats
Aykut Colakerol,Mustafa Zafer Temiz,Hasan Huseyin Tavukcu,Serdar Aykan,Sule Ozsoy,Ahmet Sahan,Engin Kandirali,Atilla Semercioz
International Journal of Impotence Research. 2020;
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82 An Overview of Ozone Therapy for Treating Foot Ulcers in Patients with Diabetes
Qing Wen,Qiu Chen
The American Journal of the Medical Sciences. 2020;
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83 Effect of ozone therapy on wound healing in the buccal mucosa of rats
Robson Pchepiorka,Maria Stella Moreira,Nelise Alexandre da Silva Lascane,Luiz Henrique Catalani,Sérgio Allegrini Jr,Nelson Batista de Lima,e Flávia Gonçalves
Archives of Oral Biology. 2020; 119: 104889
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84 Ozone: a natural bioactive molecule with antioxidant property as potential new strategy in aging and in neurodegenerative disorders
Catia Scassellati,Antonio Carlo Galoforo,Cristian Bonvicini,Ciro Esposito,Giovanni Ricevuti
Ageing Research Reviews. 2020; : 101138
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85 Efficacy of ozonated water mouthwash on early plaque formation and gingival inflammation: a randomized controlled crossover clinical trial
Alessandra Cardoso Nicolini,Isadora dos Santos Rotta,Gerson Pedro José Langa,Stephanie Anagnostopoulos Friedrich,David Alejandro Arroyo-Bonilla,Marcius Comparsi Wagner,Patrícia Weidlich,Cassiano Kuchenbecker Rösing,Juliano Cavagni
Clinical Oral Investigations. 2020;
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86 Anti-inflammatory Effect of Ozone Therapy in an Experimental Model of Rheumatoid Arthritis
Ana Paula Santos Tartari,Felipe Figueiredo Moreira,Mário César Da Silva Pereira,Emerson Carraro,Francisco José Cidral-Filho,Afonso Inoue Salgado,Ivo Ilvan Kerppers
Inflammation. 2020;
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87 A preliminary evaluation on the efficacy of ozone therapy in the treatment of COVID-19
Zhishui Zheng,Minglin Dong,Ke Hu
Journal of Medical Virology. 2020;
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88 Blood ozonization in patients with mild to moderate COVID-19 pneumonia: a single centre experience
Carlo Tascini,Giovanni Sermann,Alberto Pagotto,Emanuela Sozio,Chiara De Carlo,Alessandro Giacinta,Francesco Sbrana,Andrea Ripoli,Nadia Castaldo,Maria Merelli,Barbara Cadeo,Cristiana Macor,Amato De Monte
Internal and Emergency Medicine. 2020;
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89 Molecular mechanisms in Cognitive Frailty: potential therapeutic targets for oxygen-ozone treatment
Catia Scassellati,Miriam Ciani,Antonio Carlo Galoforo,Roberta Zanardini,Cristian Bonvicini,Cristina Geroldi
Mechanisms of Ageing and Development. 2020; : 111210
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90 Changes in Th17 cells frequency and function after ozone therapy used to treat multiple sclerosis patients
Morteza Izadi,Safa Tahmasebi,Inna Pustokhina,Alexei Valerievich Yumashev,Tayyebeh Lakzaei,Akbar Ghorbani Alvanegh,Leila Roshangar,Mehdi Dadashpour,Mehdi Yousefi,Majid Ahmadi
Multiple Sclerosis and Related Disorders. 2020; 46: 102466
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91 The right therapeutic method of ozone therapy used to treat multiple sclerosis patients
Valter Travagli
Multiple Sclerosis and Related Disorders. 2020; 46: 102545
[Pubmed] | [DOI]
92 Effect of air pollution on hospitalization for acute exacerbation of chronic obstructive pulmonary disease, stroke, and myocardial infarction
Cai Chen,Xuejian Liu,Xianfeng Wang,Wenxiu Qu,Wei Li,Leilei Dong
Environmental Science and Pollution Research. 2019;
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93 Types of microorganisms in proximal caries lesion and ozone treatment
Janet N. Kirilova,Snezhanka Z. Topalova-Pirinska,Dimitar N. Kirov,Elitsa G. Deliverska,Lilia B. Doichinova
Biotechnology & Biotechnological Equipment. 2019; 33(1): 683
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94 Ozone and pulsed electro-magnetic field therapies improve endometrial lining thickness in frozen embryo transfer cycles
Zaher Merhi,Rajean Moseley-LaRue,Amber Ray Moseley,André Hugo Smith,John Zhang
Medicine. 2019; 98(34): e16865
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95 Air Pollution and Angioedema
Suraj Kedarisetty,Evan Jones,Derrick Tint,Ahmed M. S. Soliman
Otolaryngology–Head and Neck Surgery. 2019; 161(3): 431
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96 Selected Office Based Anticancer Treatment Strategies
Jesse A. Stoff
Journal of Oncology. 2019; 2019: 1
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97 The Effect of Paravertebral Ozone Injection in the Treatment of Low Back Pain
Çagri Özcan,Ömer Polat,Haluk Çelik,Bekir Yavuz Uçar
Pain Practice. 2019;
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98 Comparative Clinical Study the Effectiveness of Plasmapheresis with Ozone Therapy in the Prevention of Ocular Herpes Recurrence
M. G. Guliyeva
Ophthalmology in Russia. 2019; 16(1): 76
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99 Effects of antibiotic and intra-peritoneal ozone administration on proinflammatory cytokine formation, antioxidant levels and abdominal organ functions in the treatment of experimentally generated infectious peritonitis in rabbits
Ozlem Guzel, Ahmet Gulcubuk, Esma Yildar, Feraye Esen Gursel, Iraz Akis, Funda Bagcigil, Ozge Erdogan Bamac, Gulay Yuzbasioglu Ozturk, Bulent Ekiz
Veterinární medicína. 2019; 64(8): 348
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100 The Biochemical and Pharmacological Properties of Ozone: The Smell of Protection in Acute and Chronic Diseases
Rosaria Di Mauro,Giuseppina Cantarella,Renato Bernardini,Michelino Di Rosa,Ignazio Barbagallo,Alfio Distefano,Lucia Longhitano,Nunzio Vicario,Daniela Nicolosi,Giacomo Lazzarino,Daniele Tibullo,Maria Gulino,Mariarita Spampinato,Roberto Avola,Giovanni Li Volti
International Journal of Molecular Sciences. 2019; 20(3): 634
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101 Ozone Therapy in a Patient with Diabetic Foot Ulcerations and a Decision for Amputation
Saltuk Aytacoglu,Barlas Naim Aytacoglu
Case Reports in Clinical Medicine. 2019; 08(02): 35
[Pubmed] | [DOI]
102 Successful treatment of acute-on-chronic liver failure and hemolytic anemia with hepato-protective drugs in combination with intravenous ozone without steroids: A case report
Zhaohui Bai,Hongyu Li,Yanqin Liu,Jiao Deng,Chunhui Wang,Yingying Li,Xingshun Qi
Intractable & Rare Diseases Research. 2018;
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103 Role of Prenatal Hypoxia in Brain Development, Cognitive Functions, and Neurodegeneration
Natalia N. Nalivaeva,Anthony J. Turner,Igor A. Zhuravin
Frontiers in Neuroscience. 2018; 12
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104 Ozone Therapy as Adjuvant for Cancer Treatment: Is Further Research Warranted?
Bernardino Clavo,Norberto Santana-Rodríguez,Pedro Llontop,Dominga Gutiérrez,Gerardo Suárez,Laura López,Gloria Rovira,Gregorio Martínez-Sánchez,Esteban González,Ignacio J. Jorge,Carmen Perera,Jesús Blanco,Francisco Rodríguez-Esparragón
Evidence-Based Complementary and Alternative Medicine. 2018; 2018: 1
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105 Molecular insights into the effect of ozone on human hemoglobin in autohemotherapy: Highlighting the importance of the presence of blood antioxidants during ozonation
Fouad Mehraban,Arefeh Seyedarabi,Zahra Seraj,Shahin Ahmadian,Najmeh Poursasan,Saeed Rayati,Ali Akbar Moosavi-Movahedi
International Journal of Biological Macromolecules. 2018; 119: 1276
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