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Management of diabetic foot ulcers (DFUs) is a great challenge for clinicians. Although the oxygen-ozone treatment improves the diabetic outcome, there are few clinical trials to verify the efficacy and illuminate the underlying mechanisms of oxygen-ozone treatment on DFUs. In the present study, a total of 50 type 2 diabetic patients complicated with DFUs, Wagner stage 2~4, were randomized into control group treated by standard therapy only and ozone group treated by standard therapy plus oxygen-ozone treatment. The therapeutic effects were graded into 4 levels from grade 0 (no change) to grade 3 (wound healing). The wound sizes were measured at baseline and day 20, respectively. Tissue biopsies were performed at baseline and day 11. The expressions of vascular endothelial growth factor (VEGF), transforming growth factor-β (TGF-β), and platelet-derived growth factor (PDGF) proteins in the pathologic specimens were determined by immunohistochemical examinations. The effective rate of ozone group was significantly higher than that of control group (92% versus 64%, P<0.05). The wound size reduction was significantly more in ozone group than in control group (P<0.001). After treatment, the expressions of VEGF, TGF-β, and PDGF proteins at day 11 were significantly higher in ozone group than in control group. Ozone therapy promotes the wound healing of DFUs via potential induction of VEGF, TGF-β, and PDGF at early stage of the treatment. (Clinical trial registry number is ChiCTR-TRC-14004415).

We used a randomized crossover experiment to estimate the effects of ozone (vs. clean air) exposure on genome-wide DNA methylation of target bronchial epithelial cells, using 17 volunteers, each randomly exposed on two separated occasions to clean air or 0.3-ppm ozone for two hours. Twenty-four hours after exposure, participants underwent bronchoscopy to collect epithelial cells whose DNA methylation was measured using the Illumina 450 K platform. We performed global and regional tests examining the ozone versus clean air effect on the DNA methylome and calculated Fisher-exact p -values for a series of univariate tests. We found little evidence of an overall effect of ozone on the DNA methylome but some suggestive changes in PLSCR1 , HCAR1 , and LINC00336 DNA methylation after ozone exposure relative to clean air. We observed some participant-to-participant heterogeneity in ozone responses.

ObjectivesTo evaluate local effect of gaseous ozone on bacteria in deep carious lesions after incomplete caries removal, using chlorhexidine as control, and to investigate its effect on pulp vascular endothelial growth factor (VEGF), neuronal nitric oxide synthase (nNOS), and superoxide dismutase (SOD).Materials and methodsAntibacterial effect was evaluated in 48 teeth with diagnosed deep carious lesion. After incomplete caries removal, teeth were randomly allocated into two groups regarding the cavity disinfectant used: ozone (open system) or 2% chlorhexidine. Dentin samples were analyzed for the presence of total bacteria and Lactobacillus spp. by real-time quantitative polymerase chain reaction. For evaluation of ozone effect on dental pulp, 38 intact permanent teeth indicated for pulp removal/tooth extraction were included. After cavity preparation, teeth were randomly allocated into two groups: ozone group and control group. VEGF/nNOS level and SOD activity in dental pulp were determined by enzyme-linked immunosorbent assay and spectrophotometric method, respectively.ResultsOzone application decreased number of total bacteria (p = 0.001) and Lactobacillus spp. (p < 0.001), similarly to chlorhexidine. The VEGF (p < 0.001) and nNOS (p = 0.012) levels in dental pulp after ozone application were higher, while SOD activity was lower (p = 0.001) comparing to those in control pulp.ConclusionsAntibacterial effect of ozone on residual bacteria after incomplete caries removal was similar to that of 2% chlorhexidine. Effect of ozone on pulp VEGF, nNOS, and SOD indicated its biocompatibility.Clinical relevanceOzone appears as effective and biocompatible cavity disinfectant in treatment of deep carious lesions by incomplete caries removal technique.

BackgroundThe gingivitis treatment protocols aim to stop the progression of inflammation and prevent the affliction of deeper periodontal tissues. Current research focuses on novel methods that can be applied with handheld and ultrasonic devices to increase the effectiveness of non-surgical periodontal treatment. Ozone has been shown to have a strong antimicrobial effect against bacteria that play a role in the etiology of gingival/periodontal diseases. Research evaluating the effects of ozone gas on the treatment of gingival tissues is limited.AimWe investigated the effects of ozone application during the scaling procedure on clinical parameters of gingivitis in smokers and non-smokers and to investigate the efficacy of ozone gas in periodontal treatment.MethodsA total of 40 subjects, 27 males/13 females—20 smokers/20 non-smokers—with plaque-induced gingivitis, were selected. The patients underwent a single session of full-mouth supragingival and subgingival scaling. Gaseous ozone application was performed on a randomly selected maxillary quadrant. The other maxillary quadrant was designated as the control group receiving only initial periodontal treatment. Clinical parameters, such as PI, GI, GBTI, and PPD, were measured before and 1 week after treatment.ResultsOur study shows that ozone therapy significantly improves clinical parameters in smokers and non-smokers when applied in addition to periodontal therapy.ConclusionsAdditional studies comparing the effect of ozone therapy on smokers and non-smokers to the effects of oral mouthwashes may provide valuable contributions to the field.

Short, high-concentration peaks of the atmospheric pollutant ozone (O₃) cause the formation of cell death lesions on the leaves of sensitive plants. Numerous similarities between the plant responses to O₃ and pathogens suggest that O₃ triggers hypersensitive response-like programmed cell death (PCD). We examined O₃ and superoxide-induced cell death in the O₃-sensitive radical-induced cell death1 (rcd1) mutant. Dying cells in O₃-exposed rcd1 exhibited several of the typical morphological characteristics of the hypersensitive response and PCD. Double-mutant analyses indicated a requirement for salicylic acid and the function of the cyclic nucleotide-gated ion channel AtCNGC2 in cell death. Furthermore, a requirement for ATPases, kinases, transcription, Ca²⁺ flux, caspase-like proteolytic activity, and also one or more phenylmethylsulfonyl fluoride-sensitive protease activities was shown for the development of cell death lesions in rcd1. Furthermore, mitogen-activated protein kinases showed differential activation patterns in rcd1 and Columbia. Taken together, these results directly demonstrate the induction of PCD by O₃.

The aim of this study was to evaluate the effect of HealOzone on two microorganisms, 4 and 8 weeks after treatment, using a tooth cavity model. Four groups of caries-free third molars ( n  = 12) were used (A, B, C and D). Three cavities were prepared into each tooth. After sterilization, groups A and B were inoculated with Streptococcus mutans , and groups C and D, with Lactobacillus casei for 48 h. One cavity of each tooth was used to evaluate the infection. After inoculation, groups B and D were treated with ozone (60 s), and groups A and C were used as controls. Then, the two cavities of each tooth were filled with composite, and the teeth were stored in sucrose medium. The restorations were removed after 4 and 8 weeks, respectively; dentin chips were collected, and the amount of microorganisms was determined. Ozone treatment reduced significantly the amount of S. mutans compared to the control group ( p  ≤ 0.05). This antibacterial effect was able to be seen after 4 ( p  = 0.0005) and 8 ( p  = 0.0002) weeks. No significant difference was found between the control and treated group as far as L. casei is concerned ( p  > 0.05). HealOzone (60 s) can provide some antibacterial treatment against S. mutans even after 8 weeks. However, an elimination of the microorganisms through HealOzone seems not to be possible. L. casei was more resistant to ozone. Although ozone exerts a significant antibacterial effect against S. mutans , it is probably not enough as the only antibacterial method, during the fillings therapy.

Objective. This study was designed to compare the effects of catheter drainage alone and combined with ozone in the management of multiloculated pyogenic liver abscess (PLA). Methods. The prospective study included 60 patients diagnosed with multiloculated PLA. All patients were randomly divided into two groups: catheter drainage alone (group I) and catheter drainage combined with ozone (group II). Drainage was considered successful when (1) the abscess cavity was drained and (2) clinical symptoms were resolved. Kruskal-Wallis nonparametric test was used to compare the success rates, length of stay (LOS), and need for further surgery of the two groups. P < 0.05 indicates significant difference. Results. All patients’ catheters were successfully placed under CT guidance. Group I was treated with catheters alone and group II was treated with catheters and ozone. The success rates of groups I and II were 86% and 96%, respectively ( P < 0.05 ). And compared with group II, the duration of fever in group I was longer ( P < 0.05 ), and the LOS was also longer ( P < 0.05 ). Conclusion. Catheter drainage combined with ozone is an effective and safe treatment in multiloculated PLA. The Clinical Registration Number is ChiCTR1800014865.

The ultimate objective of this review is to encourage a multi-disciplinary and integrated methodological approach that, starting from the recognition of some current uncertainties, helps to deepen the molecular bases of ozone treatment effects on human and animal well-being and to optimize their performance in terms of reproducibility of results, quality, and safety. In fact, the common therapeutic treatments are normally documented by healthcare professionals’ prescriptions. The same applies to medicinal gases (whose uses are based on their pharmacological effects) that are intended for patients for treatment, diagnostic, or preventive purposes and that have been produced and inspected in accordance with good manufacturing practices and pharmacopoeia monographs. On the contrary, it is the responsibility of healthcare professionals, who thoughtfully choose to use ozone as a medicinal product, to achieve the following objectives: (i) to understand the molecular basis of the mechanism of action; (ii) to adjust the treatment according to the clinical responses obtained in accordance with the principles of precision medicine and personalized therapy; (iii) to ensure all quality standards.

Objective: Dental handpieces, ultrasonic scalers, air polishers, air abrasion units produce the most visible aerosols. The objective of this study was to assess the effect of chlorhexidine (CHX), povidone iodine (PI), and ozone (OZ) on the microorganisms in dental aerosols. Materials and Methods: A total of 60 patients were included in this study, they were randomly assigned into three groups and were subjected to scaling before and after rinsing with 0.2% CHX, 1% PI or irrigation with OZ. Blood agar plates were used to collect the gravitometric settling of aerosols and were sent for aerobic and anaerobic culture. Results: The results demonstrated high percentage reduction of aerobic and anaerobic colony forming units (CFUs) in all three groups. In aerobic CFUs, CHX showed the highest reduction (57%) at mask position whereas at chest position and at 9 ft, PI showed higher CFU reductions (37% and 47%, respectively). In anaerobic CFUs, CHX showed the highest percentage of reduction at chest level (43%) and at 9 ft (44%). Conclusion: CHX, PI and OZ showed similar effects in reducing aerobic and anaerobic CFU's at the chest mask and at 9 ft. OZ can be used as a preprocedural agent, considering its beneficial effects.

Abstract Rationale Air pollution has been associated with increased prevalence of type 2 diabetes; however, the mechanisms remain unknown. We have shown that acute ozone exposure in rats induces release of stress hormones, hyperglycemia, leptinemia, and glucose intolerance that are associated with global changes in peripheral glucose, lipid, and amino acid metabolism. Objectives To examine ozone-induced metabolic derangement in humans using serum metabolomic assessment, establish human-to-rodent coherence, and identify novel nonprotein biomarkers. Methods Serum samples were obtained from a crossover clinical study that included two clinic visits (n = 24 each) where each subject was blindly exposed in the morning to either filtered air or 0.3 parts per million ozone for 2 hours during 15-minute on-off exercise. Serum samples collected within 1 hour after exposure were assessed for changes in metabolites using a metabolomic approach. Measurements and Main Results Metabolomic analysis revealed that ozone exposure markedly increased serum cortisol and corticosterone together with increases in monoacylglycerol, glycerol, and medium- and long-chain free fatty acids, reflective of lipid mobilization and catabolism. Additionally, ozone exposure increased serum lysolipids, potentially originating from membrane lipid breakdown. Ozone exposure also increased circulating mitochondrial β-oxidation–derived metabolites, such as acylcarnitines, together with increases in the ketone body 3-hydroxybutyrate. These changes suggested saturation of β-oxidation by ozone in exercising humans. Conclusions As in rodents, acute ozone exposure increased stress hormones and globally altered peripheral lipid metabolism in humans, likely through activation of a neurohormonally mediated stress response pathway. The metabolomic assessment revealed new biomarkers and allowed for establishment of rodent-to-human coherence. Clinical trial registered with www.clinicaltrials.gov (NCT 01492517).