Iacopo Zanardi

The ozone paradox: Ozone is a strong oxidant as well as a medical drug

After five decades characterized by empiricism and several pitfalls, some of the basic mechanisms of action of ozone in pulmonary toxicology and in medicine have been clarified. The present knowledge allows to understand the prolonged inhalation of ozone can be very deleterious first for the lungs and successively for the whole organism. On the other hand, a small ozone dose well calibrated against the potent antioxidant capacity of blood can trigger several useful biochemical mechanisms and reactivate the antioxidant system. In detail, firstly ex vivo and second during the infusion of ozonated blood into the donor, the ozone therapy approach involves blood cells and the endothelium, which by transferring the ozone messengers to billions of cells will generate a therapeutic effect. Thus, in spite of a common prejudice, single ozone doses can be therapeutically used in selected human diseases without any toxicity or side effects. Moreover, the versatility and amplitude of beneficial effect of ozone applications have become evident in orthopedics, cutaneous, and mucosal infections as well as in dentistry.

Ozone and Ozonated Oils in Skin Diseases: A Review

Although orthodox medicine has provided a variety of topical anti-infective agents, some of them have become scarcely effective owing to antibiotic- and chemotherapeutic-resistant pathogens. For more than a century, ozone has been known to be an excellent disinfectant that nevertheless had to be used with caution for its oxidizing properties. Only during the last decade it has been learned how to tame its great reactivity by precisely dosing its concentration and permanently incorporating the gas into triglycerides where gaseous ozone chemically reacts with unsaturated substrates leading to therapeutically active ozonated derivatives. Today the stability and efficacy of the ozonated oils have been already demonstrated, but owing to a plethora of commercial products, the present paper aims to analyze these derivatives suggesting the strategy to obtain products with the best characteristics.

Ozone acting on human blood yields a hormetic dose-response relationship

The aim of this paper is to analyze why ozone can be medically useful when it dissolves in blood or in other biological fluids. In reviewing a number of clinical studies performed in Peripheral Arterial Diseases (PAD) during the last decades, it has been possible to confirm the long-held view that the inverted U-shaped curve, typical of the hormesis concept, is suitable to represent the therapeutic activity exerted by the so-called ozonated autohemotherapy. The quantitative and qualitative aspects of human blood ozonation have been also critically reviewed in regard to the biological, therapeutic and safety of ozone. It is hoped that this gas, although toxic for the pulmonary system during prolonged inhalation, will be soon recognized as a useful agent in oxidative-stress related diseases, joining other medical gases recently thought to be of therapeutic importance. Finally, the elucidation of the mechanisms of action of ozone as well as the obtained results in PAD may encourage clinical scientists to evaluate ozone therapy in vascular diseases in comparison to the current therapies.

Ozonated sesame oil enhances cutaneous wound healing in SKH1 mice.

Ozone is well recognized as a bactericidal agent and its beneficial effect on wound healing could be a consequence of this property. Because ozone itself does not penetrate the cells but immediately reacts with polyunsaturated fatty acids, its effects should be the results of oxidative reaction. For this reason, ozonated oils could be a way to deliver ozone messengers to the skin. This paper evaluated the therapeutic effects of three different grades of ozonated sesame oil in acute cutaneous wounds made in the skin of SKH1 mice. Specifically, wound closure rate, histological parameters, and the level of key proteins such as vascular endothelial growth factors and cyclin D1 have been analyzed in relation to the peroxide level present in the ozonated oil. Treatment with moderately ozonated sesame oil—expressed as peroxide value about 1,500)—has a faster wound closure rate in the first 7 days than treatment with oil containing either lower or higher peroxide value, and even with controls. Moreover, under the same treatment, an earlier and higher response of cells involved in wound repair, a higher angiogenesis, as well as an enhanced vascular endothelial growth factors and cyclin D1 expression were observed. The present study shows the validity of ozonated sesame oil in cutaneous wound healing and emphasizes the importance of the ozonation grade.

Properties of sesame oil by detailed 1H and 13C NMR assignments before and after ozonation and their correlation with iodine value, peroxide value, and viscosity measurements

Gaseous ozone chemically reacts with unsaturated triglyceride substrates leading to ozonated derivatives with a wide potential applications, ranging from the petrochemical to the pharmaceutical industry. To date, an ultimate understanding of the ozone reactivity during sesame oil ozonation process as well as detailed (1)H and (13)C NMR assignments are lacking. A practical advantage of NMR is that a single NMR sample measurement can explain many issues, while similar analysis by traditional methods may require several independent and time-consuming measurements. Moreover, significant relationships among NMR spectra and both conventional chemical analysis and viscosity measurements have been found. Eventually, NMR could play an important role for quality attributes of ozonated oil derivatives.

How much ozone bactericidal activity is compromised by plasma components

Aims:  Evaluation of bactericidal effect of different concentrations of ozone when used (a) as a gas, or (b) dissolved in saline. The addition of hydrogen peroxide or 4‐hydroxynonenal dissolved in saline was also tested, as well as the effect of human plasma.

Effects of Ozone Blood Treatment on the Metabolite Profile of Human Blood

Metabonomic characterization of the effects caused by ozone and other stressors on normal human blood was performed. Samples of blood obtained from healthy subjects were treated ex vivo with increasing concentrations of ozone and/or with UV radiation and heat. 1H-NMR analysis of plasma samples after treatments showed the quantitative variation of some metabolites and the formation of new metabolites normally absent. Both the increment of some metabolites like formate, acetoacetate, and acetate and the decrement of pyruvate were of particular interest. Moreover, the oxidation of ascorbic acid and the transformation of uric acid into allantoin after ozonation within the therapeutic concentration range were observed. In the ozonated spectra, 2 unidentified peaks appeared at 2.82 ppm and 8.08 ppm. They are related to the direct antioxidant activity of albumin in the presence of ozone and they could be considered as specific markers of the blood ozonation.

Ozone: A New Therapeutic Agent in Vascular Diseases

In this article, we scientifically evaluate the bio-oxidative procedure known as oxygen-ozone therapy. Research over a decade has established a comprehensive framework for understanding and recommending this type of autohemotherapy in vascular diseases. In contrast, a non-specific immunomodulation therapy, using heavily oxidized and denatured blood, has been recently used in studies involving a total of approximately 3000 patients and has led to ‘disappointing’ results. Such a treatment appears to be an inappropriate example of the so-called minor autohemotherapy, and its poor outcomes may discourage any further studies. Therefore it appears necessary to clarify that the use of only a minimal ozone dose and a valid experimental protocol is likely to produce beneficial results. Millions of people suffer from chronic limb, brain, and heart ischemia, and such patients may benefit if appropriate ozone therapy could be implemented. Accordingly, we propose the need for a well designed, multicenter, clinical trial to be conducted.

Topical Applications of Ozone and Ozonated Oils as Anti-Infective Agents: An Insight into the Patent Claims

Orthodox medicine has been very active in the field of topical anti-infective agents and current chemotherapy has procured valid antibiotics, antivirals, vaccines, antibodies, and antiparasitic drugs to be parenterally and/or topically used. However, these drugs may cause side effects and sometimes provide unsatisfactory results because pathogens become drug-resistant. Another drawback is represented by their cost, which compromise their use or their availability in poor Countries. Therefore, there is a critical need for new strategies to treat dermatological affections. The old intuition for using ozone in the treatment of necrotic wounds, especially if due to anaerobic bacteria, is now justified by the studies about reactive oxygen species generation by granulocytes and macrophages as the first line of defense during an infection. As a consequence, the disinfectant value of ozone has been increasingly appreciated during the last fifteen years. This review summarizes the patents filed and issued, with particular emphasis to the more recent patents, about the anti-infective topical use of ozone: i) in the gaseous form; ii) after gaseous ozone saturation of aqueous, not-oily pharmaceutical vehicles and solvents; iii) where gaseous ozone chemically reacts with unsaturated substrates leading to therapeutically active ozonated derivatives. We hope that recent advances and a better understanding of the ozone chemistry and biology will be able to create the mental attitude to prove the validity of large-scale therapeutic use of both ozone and ozone derivatives as topical anti-infective agents by performing multicenter, randomized clinical studies, as aptly requested by orthodox medicine.

Physico-Chemical Characterization of Sesame Oil Derivatives

Ozone treatment of commercially available vegetable oils gives rise to the formation of chemical species that are responsible for the therapeutic properties of ozonated oil derivatives in dermatological diseases. In the last years, these products have been successfully used as a topical disinfectant in a number of serious skin affections. The medical application of empirically prepared ozonated oil has yielded striking improvements with unexpected and rapid healing, compelling us to begin a long-range study aiming first to define the main characteristics of the most common ozonated vegetable oils, about which there is usually no medical consensus because of the lack of standardization of their technological parameters. Sesame oil was selected because of its great amount of polyunsaturated acyl groups, as well as natural antioxidants. Moreover, we have determined the kinetics and optimal conditions of ozonation (e.g., ozone concentrations, time of exposure, temperature) for obtaining an ozonated oil characterized by well-established technological and physico-chemical properties, namely an accurate peroxide value determination. On the basis of the results, we have gained an understanding of the modifications of the vegetable oils during the ozonation process.