Ozone mediators effect on "in vitro" scratch wound closure.

The beneficial effect of low doses of ozone on wound healing has been well documented and attributed mainly to its bactericidal and pro-oxidant properties. Because ozone itself does not penetrate the cells but immediately reacts with polyunsaturated fatty acids, its effects are the results of oxidative mediators. Among the molecule produces by the interaction of ozone with biological systems, there are HNE and H2O2. At today, the cellular mechanisms accounting for the positive effects of mild ozonization on wound closure are still largely unexplored. The aim of the present study was to evaluate the effect of different non-toxic doses of ozonated saline ranging from 2 to 300 µM, in an in vitro wound scratch model by the use of human keratinocytes. The results showed that ozonated saline is able to improve in vitro wound healing by stimulating cell proliferation as measured by BrdU assay and PCNA protein levels. In order to better elucidate the molecules that play the main role in the beneficial effect of ozonated saline in wound healing, HNE and H2O2 were used alone or in combination to mimic ozonated saline effect. Surprisingly, keratinocytes treated with different doses of HNE and H2O2 did not significantly improve the wound closure, while the combination of the two compounds was able to improve wound closure. In addition, Nrf2 pathways were also activated as determined by its translocation to the nucleus and the increased HO1 gene expression. The present work suggests that ozonated saline effect on wound closure is the results of the combination of more molecules among which HNE and H2O2 play a key role.

The usefulness of ozone treatment in spinal pain.

OBJECTIVE: The aim of this review is to elucidate the biochemical, molecular, immunological, and pharmaceutical mechanisms of action of ozone dissolved in biological fluids. Studies performed during the last two decades allow the drawing of a comprehensive framework for understanding and recommending the integration of ozone therapy for spinal pain. METHODS: An in-depth screening of primary sources of information online - via SciFinder Scholar, Google Scholar, and Scopus databases as well as Embase, PubMed, and the Cochrane Database of Systemic Reviews - was performed. In this review, the most significant papers of the last 25 years are presented and their proposals critically evaluated, regardless of the bibliometric impact of the journals. RESULTS: The efficacy of standard treatments combined with the unique capacity of ozone therapy to reactivate the innate antioxidant system is the key to correcting the oxidative stress typical of chronic inflammatory diseases. Pain pathways and control systems of algesic signals after ozone administration are described. CONCLUSION: This paper finds favors the full insertion of ozone therapy into pharmaceutical sciences, rather than as either an alternative or an esoteric approach.

Spectroscopic characterization of both aqueous and solid-state diacerhein/hydroxypropyl-ß-cyclodextrin inclusion complexes.

Diacerhein, a poorly water soluble antirheumatic prodrug, was spectroscopically characterized to form inclusion complexes with hydroxypropyl-ß-cyclodextrin (HPßCD) in both aqueous solution and in solid phase. Complexation with the hydrophilic carriers was used to improve the solubility and dissolution rate of the compound. The kinetics of the prodrug degradation to the active rhein in aqueous buffer solution were also investigated as a function of HPßCD concentration. The solid complexes prepared by different methods such as physical mixture, kneading, co-evaporation method and freeze dried method in 1:1M ratio, were characterized by DSC and FTIR. The dissolution profiles of solid complexes were determined and compared with diacerhein alone and their physical mixture, in the simulated intestinal fluid at 37°C. The accurate molecular spectroscopic characterization of diacerhein in the presence of different amounts of aqueous cyclodextrins was essential to determine the correct binding constants for the diacerhein/HPßCD system. The binding constants were also validated by UV spectrometry and HPLC procedure in order to compare the values from the different methods. Higuchi-Connors phase solubility method has proved not suitable when either the free or/and the complexed prodrug degrade in aqueous solution.

An integrated medical treatment for type-2 diabetes.

This paper tries to emphasize two relevant concepts: the first is that type 2 diabetes is a chronic diseases characterized by both a dysmetabolism and a chronic oxidative stress. A variety of orthodox drugs are somewhat able to correct the metabolic alterations, but do not deal with the chronic inflammation. Consequently, as the validity of precisely treating blood with therapeutic ozone concentrations in restoring a redox homeostasis has been now demonstrated, the integration of ozone therapy appears essential for a rational treatment of type 2 diabetes. Such a combination may be able to reduce the diabetic epidemic.

Emerging topics in cutaneous wound repair.

The intervention strategies in various types of skin wounds include several treatment programs that depend on the identified disease. Several factors such as aging, defective nutrition, traumatism, atherosclerosis, and diabetes may contribute to the formation of a wound that has no tendency to heal due to a defective and complicated repair process. The numerous advances in the understanding of the wound-healing process in both acute and chronic lesions have been recently described. The purpose of this paper is to describe relatively new approaches as viable alternatives to current wound-healing therapies. The future challenges for both the best targeting and optimization of these potential treatments are also described.

Reliable and effective oxygen-ozone therapy at a crossroads with ozonated saline infusion and ozone rectal insufflation.

OBJECTIVES: This review aims to highlight the advantages and safety of oxygen-ozone therapy (OOT) and to suggest ways to enhance its acceptance. KEY FINDINGS: The treatment of a herniated disk by injecting a gaseous oxygen-ozone mixture inside the nucleus pulposus is a great clinical success. However, the use of OOT lags for a number of reasons, including lack of standardization, the need for numerous treatments, lack of knowledge and even denial. Anecdotally, several million treatments by OOT have been performed worldwide indicating its usefulness, mainly in peripheral arterial diseases and age-related macular degeneration. The scepticism that accompanies the systemic use of ozone can only be overcome by demonstrating the validity of OOT in controlled and randomized clinical trials. Cheaper and quicker methods, such as ozonating physiological saline with successive infusion as well as ozone rectal insufflations, are becoming popular, however, such alternative procedures are erratic, unstable and liable to be toxic, with deleterious consequences, and are likely to discredit the beneficial use of ozone. SUMMARY: The approval of ozone in terms of both therapeutic efficacy and safety will depend on the results achieved by authoritative clinical trials.

Selective ozone concentrations may reduce the ischemic damage after a stroke.

Abstract Stroke is one of the most debilitating diseases, and it is unfortunate that only a small percentage of patients can be treated with thrombolytic agents. Consequently, there is an urgent need of finding an alternative procedure for reoxygenating the so-called penumbra at the earliest time as possible for reducing morbidity and disability. A preliminary, preclinical study has been carried out by using rat hippocampal and cortical brain slices subjected to oxygen-glucose deprivation. Oxygen-ozone gaseous mixture appeared to be effective in reverting damage of brain tissues, supporting the evaluation of this approach in well-designed clinical trials in stroke patients.

Oxygen/ozone as a medical gas mixture. A critical evaluation of the various methods clarifies positive and negative aspects.

Besides oxygen, several other gases such as NO, CO, H2, H2S, Xe and O3 have come to age over the past few years. With regards to O3, its mechanisms of action in medicine have been clarified during the last two decades so that now a comprehensive framework for understanding and recommending ozone therapy in various pathologies is available. O3 used within the determined therapeutic window is absolutely safe and more effective than golden standard medications in numerous pathologies, like vascular diseases. However, ozone therapy is mostly in practitioners' hands and some recent developments for increasing cost effectiveness and speed of treatment are neither standardized, nor evaluated toxicologically. Hence, the aim of this article is to emphasize the need to objectively assess the pros and cons of oxygen/ozone as a medical gas mixture in the hope that ozone therapy will be accepted by orthodox medicine in the near future.

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.

Diabetes and chronic oxidative stress. A perspective based on the possible usefulness of ozone therapy.

It is now well established that hyperglycemia, present in both type 1 and type 2 diabetes, causes a variety of biochemical derangements leading to a diffused vascular damage responsible for several pathologic manifestations. Although preclinical and clinical studies have been performed by an unreliable administration route, the correct approach of oxygen-ozonetherapy may break a vicious circle. Messengers, released by a precise interaction ex vivo of the patient's blood with an equivalent calculated dose of ozone (0.42-0.84 mM), react with a variety of cells after blood infusion and restore a number of functions went astray. This paper aims to open a debate on this new therapy for improving the prognosis of diabetes.

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.

Ozonation of human HIV-infected plasmas for producing a global vaccine: How HIV-patients may help fight the HIV pandemia.

A vaccine against HIV able to generate properly neutralizing antibodies and an efficacious cytotoxic T lymphocyte response is of paramount importance. We are proposing a novel approach based on the collection of thousand small HIV-infected human plasma samples for preparing a global vaccine, able to counteract HIV diversity and mutagenicity. The pooled plasmas will undergo several steps for sterilizing and inactivating HIV, possibly other contaminant viruses and other pathogens. The critical step is the prolonged and controlled exposure of plasmas to ozone so that finally each ml of plasma has interacted with a precise dose of ozone. To inactivated plasma, both therapeutic human albumin and ozonated ethyl oleate are added for enhancing a proficient absorption and reaction with the immune system of the vaccine. The need of a partner collaboration for developing the production and the preliminary testing of the vaccine is essential.

Potentiality of oxygen-ozonetherapy to improve the health of aging people.

During the last century the lifespan of human beings has increased from about 49 to almost 80 years owing to the great advances of biomedical sciences. This fact has strongly stimulated the idea that it may be possible to prolong productive life for another twenty years but paradoxically the problem of both hyper- and hyponutrition severely jeopardizes this objective all over the world. Many causes of premature aging have been discussed in order to prevent it or find suitable medical treatment. So far only a moderate restriction of caloric intake that does not alter essential nutrients has proved capable of keeping animals and humans healthier for a prolonged time. This modality appears to activate critical longevity genes that can prolong survival: although this is a valuable line of research it will take considerable time to produce valid drugs to selectively activate these relevant genes. Meantime we propose to evaluate an easy and well accepted treatment based on a weekly quasi-total body exposure to oxygen-ozone inside a thermostatically controlled cabinet. The rational exposure to a minimal amount of ozone acting as a mild stressor induces a striking improvement of crucial metabolic activities capable of preserving a good health for several years.

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. (1)H-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.

Are dialysis devices usable as ozone gas exchangers?

A study aimed to compare the efficiency of the ozone transfer of four hydrophilic dialysis filters, and one hydrophobic gas-exchange device (GED) has been performed. Obviously, the former should be specifically used only for dialysis. Unfortunately, some clinicians incautiously use them as GEDs. It has been shown that: (i) dialysis filters present a wide range of gas-exchange yield (from 0 up to 70%), often related to variability according to the treatment time; (ii) by scanning microscopy, it has been noticed that hollow fibers are somewhat altered by ozone; and (iii) because their constitutive materials may not be ozone-resistant, they may release toxic compounds harmful for the patients. On the contrary, the appropriate GED is ozone-transfer efficient, is ozone-resistant, and is suitable for blood autotransfusion and ozonation.

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.