Ozone (O3) was discovered earlier through oxygen electrolysis and is considered the most potent natural oxidant as triatomic oxygen. Its use in water purification dates back to the late 1800s, with the first literature publication on its medical use titled “Ozone” dating back to 1885 [1].

During World War II, it was used medicinally in various instances, including intrathecal use for meningitis [2]. As a molecule of triatomic oxygen, O3 is an extremely effective oxidant. Two independent research groups discovered the major biochemical modulation induced by ozone.

Among these effects are primarily mentioned:

  • the increase in red blood cell diglycerophosphate[3];
  • improvement of red blood cell rheological properties and endothelial nitric oxide production;
  • greater difference in arterial/venous partial pressure, indicating increased mitochondrial oxygen consumption;
  • modulation of the immune system and reduction of inflammation;
  • improvement of the antioxidant status through antioxidant enzymes (including superoxide dismutase) and glutathione in cells. Effects can be obtained through O3 generating redox signalling molecules called ozonides and other reactive oxygen species with shorter lifespans (peroxides, aldehydes, etc.). Other studies also identify significant “antibiotic” activity.
Molecular Structure,Ozone.

Preconditioning with ozone in rats, without any other treatment, improved their survival by 62.5% after a lethal injection of faecal material into the peritoneum[4]. In another study, intraperitoneal preconditioning in combination with antibiotic therapy synergistically improved survival by up to 93% [5]. In endotoxic shock, such preconditioning also occurred with steroids in reducing the release of tumour necrosis factor alpha [6]. O3 has been reported to prevent drug resistance in humans by Mycobacterium tuberculosis [7]. O3 may be the ideal therapy for viral pathologies. To successfully penetrate cells, many viruses require membrane glycoproteins in reduced form RSH rather than oxidized (R-S-S-R). Therefore, reducing sulfhydryls seems necessary for the virus to evade neutralizing antibodies, as with the Ebola virus [8]. A study highlighted that if thiol groups are oxidized, cytomegalovirus loses infectivity, while when thiols are chemically reduced, the virus regains 65% of infectivity. Sampling of viruses inactivated by ozone thus includes cytomegalovirus, human immunodeficiency virus, Norwalk, MS2 bacteriophage, hepatitis A, and poliovirus [1]. In a recent study, Izzotti et al. demonstrated the effectiveness of a new treatment based on ozonized oil (HOO) taken orally for chemoprophylactic purposes in healthy subjects for COVID-19 prevention and, at high doses, for therapeutic purposes in infected patients [9].

In an even more recent study [10], the same authors have demonstrated the effectiveness of complementary therapy based on the intake of HOO for cancer treatment: in vitro, in lung and glioblastoma tumour cells, ozonized oils with a high content of ozonides suppressed the vitality of tumour cells by activating mitochondrial damage, intracellular calcium release, and apoptosis. In vivo, a total of 115 oncological patients (aged 58 ± 14 years; 44 males, 71 females) were treated with ozonized oil as a complementary therapy in addition to standard chemo/radiotherapy regimens for up to 4 years. Glioblastoma, pancreatic adenocarcinoma, cutaneous epithelioma, small and non-small cell lung carcinoma, colon adenocarcinoma, breast cancer, and prostate adenocarcinoma were included; the survival rate was significantly improved in oncological patients who received HOO as integrative therapy compared to those treated with standard therapy alone. Ozonized oils are composed of unsaturated fatty acids that have been subjected to ozone action. Ozone adds to carbon-carbon double bonds, forming ozonides. These molecules rapidly reorganize according to the Criegee mechanism, causing the formation of trioxanes. Ozonides are generally unstable, while trioxanes are relatively stable but decompose THE OZONE3 MOLECULE under the action of reducing agents or intracellular enzymes. When ozone is added to the oil
saturates, the double bonds and viscosity increase with progressive ozonide formation until the oil reaches a jelly-like consistency. Peroxides in the oil can hydrolyze to form aldehydes and ketones with shorter chains than the original fatty acid. The length of the residue is determined by the position of the double bond along with the reacting chain with ozone

In a recent study [11] aimed at evaluating the pharmacokinetics of the oxidative effects induced in the blood plasma after low-dose ozonized oil (HOO) administration, it was demonstrated that HOO can appreciably and objectively modulate the oxidative state of the treated subject. This
study shows that the increase in oxidative burden is visible 2.5 hours after HOO intake and is detectable by the increase in hydrogen peroxide equivalents. The body reacts by activating its antioxidant defences. This response was documented by the progressive increase in ascorbic acid equivalents observed starting from 9 hours after HOO intake. This rebound effect persisted and intensified up to 23 hours after administration, reaching values well above those observed before treatment. Activation of antioxidants induced by low-dose HOO treatment is clearly visible from the assessment of the total antioxidant to oxidant ratio. This ratio decreases up to 5 hours after treatment initiation and then steadily rises in subsequent times, reaching values over 46% higher than those observed before treatment initiation. It is interesting to note that this increase is not transient but stable, still well observable 23 hours after HOO intake. The modulation exerted by HOO on plasma antioxidants is a response based on the activation of defence mechanisms –
specifically antioxidants – by a noxa quantitatively below the threshold compared to the defence mechanisms themselves. The result of this interaction is the activation of defence mechanisms in the absence of biological damage [12]. Therefore, HOO administration would naturally activate the antioxidant response aimed at neutralizing oxidizing species with consequent activation of antioxidants that increase after 9 hours of HOO intake. This latency period is necessary for the body to activate the Antioxidant Responsive Elements (ARE) system represented by genes encoding antioxidant enzymes and oligopeptides (e.g., reduced glutathione composed of the amino acids glutamic acid, cysteine, and glycine), as the production time of messenger RNA by these genes and its translation into proteins at the ribosomal level is approximately 8 hours. This physiological situation explains the chronological kinetics observed in the antioxidant rebound induced by low-dose HOO. This data also demonstrates that the antioxidant response
to HOO is not due to the simple mobilization of endogenous antioxidants already present but to the production of new antioxidant proteins, which by their nature have very long half-lives. Therefore, the benefit gained from the administration of HOO appears to be stable over time.

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