Ozone-treated Group Research Articles

The impact of ozone therapy on the progression of COVID-19 patients

Introduction Ozone therapy can be used as a monotherapy or as an adjunctive treatment to standard COVID-19 treatment protocols. Current evidence indicates that this approach may improve clinical outcomes, paraclinical markers, and reduce radiological signs of inflammation, with no side effects. Material and methods he study included 100 consecutive patients aged 18 and older with COVID-19, admitted to the Intensive Care Unit at the Institute of Emergency Medicine. Patients were randomly divided into two groups: 50 patients underwent treatment according to the National Clinical Protocol along with major ozonated autohemotherapy (the study group), while the other 50 patients were treated only according to the National Clinical Protocol (the control group). Results Although the initial oxygenation index (PaO2/FiO2) values were similar in both study groups, a dynamic analysis revealed a clear efficacy of ozone therapy. By the end of the first-week treatment, the mean oxygenation index in the ozone-treated group was significantly higher than in the standard treatment group: 296.8±105.1 mm Hg versus 232.8±110.6 mm Hg (p<0.01). The use of oxygen therapy (70.0% vs. 78.0%), non-invasive ventilation (70.0% vs. 76.0%), and invasive mechanical ventilation (22.0% vs. 38.0%) tended to be lower in the ozone group, though this difference was not statistically significant (p>0.05). Both treatment groups showed a significant clinical improvement, with 54.0% of COVID-19 patients in the ozone group and 50.0% in the conventional treatment group achieving a two-point reduction in clinical severity score (p>0.05). Conclusions The mean oxygenation index significantly increased in the study patient group (246.86±30.3 mm Hg on day 1 and 296.75±105.1 mm Hg on day 7 of treatment; p<0.01) and remained unchanged in the control group (235.86±33.4 mm Hg on day 1 and 232.82±110.6 mm Hg on day 7 of treatment; p>0.05). Although the mortality rate was lower among COVID-19 patients treated with ozone therapy (24.0%) compared to those receiving standard treatment (34.0%), this difference did not achieve any statistical significance.

Analysis of the Antioxidant Mechanism of Ozone Treatment to Extend the Shelf Life and Storage Quality of ‘Korla’ Fragrant Pears Based on Label-Free Proteomics

In this study, the physiological indicators, reactive oxygen species (ROS) levels, and activities and expressions of key enzymes related to ROS metabolism were monitored to explore the mechanism of ozone treatment on the shelf life of postharvest Korla fragrant pears. The results show that postharvest fragrant pears treated with ozone had a higher firmness and lower weight loss rate and decay rate during their shelf life, especially in the late stage. Ozone treatment could also delay the occurrence of the respiratory peak and reduce the peak value. The generation rate of superoxide anion (O2−), the hydrogen peroxide (H2O2) content, and the malondialdehyde (MDA) level were reduced in the ozone-treated group, while the activities of key enzymes, superoxide dismutase (SOD), catalase (CAT), and peroxidase (POD), related to ROS metabolism in postharvest fragrant pears were stimulated by ozone treatment, especially in the middle and late stages of shelf life. Analysis of the proteomics results indicates that the POD family enzymes were the main target protein in postharvest fragrant pears treated by ozone during the middle and late stages of shelf life. The activity and expression of antioxidant-related enzymes in postharvest fragrant pears were stimulated by ozone to accelerate the metabolism of ROS and maintain high quality, especially in the middle and late shelf lives.

Effects of Ozone Treatment on Postharvest Mucor Rot of Codonopsis pilosula Caused by Actinomucor elegans

Fresh Codonopsis pilosula is highly susceptible to fungus contamination during post-harvest storage, which not only compromises the quality of C. pilosula but also contributes mycotoxin contamination, posing a significant threat to human health. Studies have indicated that ozone treatment can inhibit post-harvest diseases in fruits and vegetables. The impact of ozone treatment on the disease incidence, disease severity index, and weight loss rate of the fresh C. pilosula infected with Actinomucor elegans was investigated through the spray inoculation of A. elegans on C. pilosula tissues. Changes in the main active ingredients of C. pilosula after ozone treatment were analyzed, and the effects of ozone treatment on the integrity of cell membranes in C. pilosula tissue and reactive oxygen species (ROS) metabolism were studied. The results showed that ozone treatment had a significant inhibitory effect on the A. elegans-induced mucor rot in C. pilosula, significantly reducing the incidence of the disease. Compared with the control group, the ozone-treated group maintained the effective components of C. pilosula well. Furthermore, ozone treatment reduced the cell membrane permeability and Malondialdehyde (MDA) content in C. pilosula, significantly increased the activity of antioxidant enzymes in the ROS metabolism pathway, prevented oxidative stress caused by the accumulation of ROS in C. pilosula tissues, and maintained the integrity of cell membranes.

Effectiveness of ozone in the management of chronic periodontitis: A clinico-microbiological study

Aim: To evaluate the efficacy of ozonated water with NSPT compared with scaling and root planing (SRP) alone in treating chronic periodontitis. Settings and Design: The current study is a double-masked, randomized clinical trial. Methods and Material: A total of 30 patients aged 25 to 55 years were divided randomly into two groups having probing pocket depths of ≥ 5 mm. Full mouth SRP were performed in group 1 followed by additional sub-gingival irrigation of ozonated water in group 2. Assessment of plaque index (PI), gingival index (GI), probing pocket depth (PPD), and clinical attachment levels (CAL) was performed at baseline and six months. Microbiologic assessment with real-time polymerase chain reaction was performed for Dialister pneumosintes and Micromonas micros by collecting plaque samples. Statistical Analysis Used: The data were subjected to statistical analysis with IBM SPSS Software version 21. The results were presented under the headings of various parameters considered for this study. Results: The results revealed that there was a reduction in PI, GI, PD, CAL, and microbiologic parameters in both groups following SRP; the results were more significant in group 2 treated by sub-gingival irrigation with ozonated water, when compared with SRP alone in group 1. Also, it was noted that there was a statistically significant reduction of M. micros compared to D. pneumosintes in the ozone-treated group. Conclusions: The Subgingival irrigation of ozonated water in conjunction with SRP has beneficial effects in treating patients with chronic periodontitis.

Ozone treatment combined with sodium fluoride for the treatment of dentin hypersensitivity: an exploratory study.

Dentin hypersensitivity is one of the most common complains faced by dental professionals, and there is still lack of consensus for the most effective treatment approaches. The aim of the present study was to evaluate the efficacy of ozone therapy combined with sodium fluoride in reducing pain associated with dental hypersensitivity, compared to a paint-on adhesive desensitizing agent. The study included 20 patients (131 teeth), selected according to specific inclusion criteria and divided into two groups according to the type of desensitizing therapy (ozone with sodium fluoride or paint-on adhesive). The efficacy of both treatments were evaluated recording pain for each treated tooth immediately after treatment, after 1 month and after 6 months using a Numeric Rating Scale (NRS). Subjective parameters were also recorded: patients were asked to report whether they experienced pain caused by air, teeth brushing, cold and hot topical stimuli (present/absent). In both groups, pain values significantly decreased over time. Six months after treatment we registered significantly lower pain values in the ozone-treated group compared with the adhesive group and the mean percentage of NRS reduction from was significantly higher in the ozone-treated group. We did not register significant differences in subjective pain-related parameters between groups. The results of this exploratory study were aimed at exploring the combined effect of ozone with natrium fluoride, which was before described only in vitro. With the limitation of the sample size, our results suggest that this approach provides a pain reduction that was comparable to the paint-on adhesive.

Mechanism of ozone alleviation of malignant ascites in hepatocellular carcinoma through the inhibition of neutrophil extracellular traps.

Malignant ascites in hepatocellular carcinoma is usually a sign of advanced disease and poor prognosis and is also thought to be associated with chronic inflammation mediated by neutrophil extracellular trap (NET) networks. Although ozone, a strong oxidant, has significant antibacterial and anti-inflammatory effects, its effectiveness in treating malignant liver ascites is unclear. We first measured the levels of NETs in the peripheral blood of patients with liver cancer and healthy individuals. Next, we constructed the H22 tumor-bearing mouse model and observed the abdominal girth, body weight, survival rate, and survival time in each group; we marked the proteins associated with NETs in mouse intestinal tissues by immunofluorescence; cf-DNA and cytokines in ascites such as: tumor necrosis factor alpha (TNF-α), vascular endothelial growth factor (VEGF), interleukin 6 (IL-6), matrix metalloprotein 9 (MMP-9), and interferon gamma (IFN-γ) levels in ascites were measured by enzyme-linked immunosorbent assay. The expression levels of phosphorylated adenylate-activated protein kinase (P-AMPK) and scavenger receptor-A (SR-A) were detected by immunocytochemistry in the intestinal tissues of each group of mice. We further examined the expression of P-AMPK and SR-A proteins in ascites deposits by Western blotting. The results show, the plasma levels of NETs were higher in patients with hepatocellular carcinoma than in normal subjects (P < 0.01). Abdominal girth and body weight were significantly reduced in the ozone-treated group compared with the model group, while survival and survival time were dose dependently increased (both P < 0.05). NET-associated guanine histone H3 and myeloperoxidase were abundantly expressed at neutrophil aggregates in the intestinal tissues of the model mice, whereas their expression was significantly reduced in the ozone-treated group. The levels of cf-DNA, IL-6, IFN-γ, MMP-9, VEGF, and TNF-α were dose dependently increased in the ascites of H22 tumor-bearing mice in the ozone-treated group compared with the model group (all P < 0.01), while the expression of P-AMPK and SR-A proteins was increased in the ozone-treated group compared with the model group. Ozone showed significant antiperitoneal fluid production properties in H22 tumor-bearing mice, and ozone reduced peritoneal fluid production by activating AMPK and up-regulating SR-A phagocytosis damage-associated molecular patterns to reduce the production of NETs. This suggests that ozone could be used as a new drug for the treatment of malignant ascites in hepatocellular carcinoma.

Gaseous ozone regulates reactive oxygen species metabolism and ascorbate–glutathione cycle to delay the senescence of fresh-cut red pitaya (Selenicereus undatus) fruit

Mechanical damage during processing can accelerate the senescence and deterioration of fresh-cut red pitaya (Selenicereus undatus), thereby affecting its nutritional quality and commodity value. In this study, the effects of a low-concentration and long-term (10 μL L−1 for 60 min) gaseous ozone treatment on the quality, reactive oxygen species (ROS) metabolism, and enzyme and non-enzyme antioxidant activities of fresh-cut red pitaya fruit were investigated. The results showed that ozone treatment delayed the decline in fruit firmness, total soluble solids, and titratable acid content. Ozone treatment also maintained a higher free radical scavenging capacity and lower membrane lipid peroxidation than the control treatment. It prevented the rapid increase in ROS level by improving the activity of antioxidant enzymes, including superoxide dismutase, peroxidase, and catalase. Additionally, the treatment activated the ascorbate–glutathione cycle. Compared with the control group, ozone treatment maintained the activities of ascorbic acid peroxidase, dehydroascorbic acid reductase, monodehydroascorbic acid reductase, and glutathione reductase. Although the contents of ascorbic acid (AsA) and dehydroascorbic acid (DHA) decreased within 10 days of storage, the AsA/DHA and glutathione/oxidized glutathione ratios in the ozone-treated group were higher than those in the control group from day 0 to day 6. Thus, ozone treatment regulated ROS metabolism, enhanced the ROS-scavenging system, maintained the redox balance of fresh-cut red pitaya, and delayed senescence.

Effects of ozone therapy in an experimental murine model of Candida albicans vulvovaginitis

The antifungals used in the treatment of vulvovaginal candidiasis, although effective, can have side effects and high cost, which may not occur with the medicinal use of ozone. The study aimed to evaluate the effects of medicinal home ozone therapy on C. albicans. The experiment was carried out in vitro, cultivating C. albicans in 10mm Petri dishes, containing solid Agar Sabour and Dextrose (Oxoid). The in vivo experiment used 45-day-old female C57/BL6 mice and after suppression of ovarian activity, 20 µL of C. albicans suspension in sterile PBS, at a concentration of 105 yeast cells, were inoculated in a single dose under anesthesia. Distribution occurred in 4 groups: (i) Absolute Control Group (GCA): Not subjected to infection by the pathogen. (ii) Control Group (CG): Submitted to C. albicans infection, but not treated. (iii) Ozone Therapy Group (GO): Subjected to infection and treated with medicinal ozone therapy. (iv) Lavage Group (GL): Subjected to infection and subsequently treated with vaginal lavage. It was found that GC had a higher amount of CFU when compared to GO. Exposure for 600s was able to reduce the number of CFUs by 98.89% when compared to GC. The study concludes that ozone gas showed great efficacy in the proliferation of C. albicans in vitro. The best results were observed with application in liquid medium, however, ozone therapy was able to significantly reduce infection in the experimental model used. In in vivo experiments, the number of CFU/ml found was lower in the ozone-treated group.

Effect of Ozonated Water on Oral Mucositis and Pain Induced by Head and Neck Radiotherapy: A Cross-sectional Study

Background: Oral mucositis and local pain are the negative effects of the treatment of head and neck cancer with radiotherapy. Objectives: The study aimed to investigate the effect of ozonated water on the severity of mucositis and pain in (1) patients who were treated with ozonated water and (2) patients who were not treated with ozonated water. Methods: We randomly divided 93 patients with head and neck malignancy (aged 18 to 80) into three groups. Ozone-treated group 1 rinsed their mouth with 15 mL of ozonated water with a concentration of 20 - 50 ppm from the first session of radiotherapy for one minute before and after each session. Ozone-treated group 2 rinsed their mouth with 15 mL of ozonated water with a concentration of 20 - 50 ppm for three minutes and then swallowed it before and after each session. Ozone-treated groups 1 and 2 and the non-ozone-treated group received standard treatment if mucositis symptoms appeared in each patient. The minimum number of radiotherapy sessions was 30, and the minimum planned dose for each patient was 50 Gray. Anamnesis and the following clinical parameters were taken: the degree of mucositis, the use of corticosteroids, radiotherapy method, radiation dose, and Pain Visual Analog Scale. Multi-level and subgroup analyses were performed on the ozone-treated and non-ozone-treated levels. Results: The mean degrees of oral mucositis and pain were lower in the ozone-treated group 2 than in the ozone-treated group 1 and non-ozone-treated group (P &lt; 0.05). The non-ozone-treated group had the highest degrees of oral mucositis and pain severity (P &lt; 0.05). The Kruskal-Wallis H test showed that there was a statistically significant difference in the Visual Analog Scale of sessions 5, 10, 15, 20, 25, and 30 between different groups. However, there was no statistically significant difference in the Visual Analog Scale of session 1 (χ2 (2) = 1.022, P = 0.6). This study revealed that ozonated water can be used for preemptive pain control and mucositis. This finding aligned with previous studies. Also, former research proved the safety and efficacy of ozonated water in dentistry and medical uses. Conclusions: The use of ozonated water in patients with head and neck malignancy can reduce the pain severity and oral mucositis induced by radiotherapy. It seems that ozonated water can be used as a preemptive agent in patients who receive head and neck radiotherapy.

Consequences of continuous ozonation on the health and welfare of Atlantic salmon post-smolts in a brackish water recirculating aquaculture system.

This study investigated the biological consequences of 45-day continuous ozonation on Atlantic salmon (Salmo salar) post-smolts in a brackish water recirculating aquaculture system (RAS). There was no significant difference in survival, operational welfare indicators, and average weight at termination between the ozone-treated and control groups. Plasma biochemical analyses revealed that the creatinine level was significantly higher in the ozone-treated group than in the control at termination. Histological evaluation of skin health showed no significant difference between the two groups. On the other hand, quantitative histopathology disclosed that the ozone group exhibited a better gill health status than did the control group, particularly at the end of the trial. Mucosal transcriptomics revealed a distinct response profile between the gills and skin. At day 45, there were no differentially expressed genes (DEG) identified in the skin, in contrast to 242 ozone-induced DEGs in the gills. Assessment of the transcriptomic profiles over time revealed that temporal effects were of greater impact in skin compared to gills, regardless of the treatment. The treatment did not result in metabolomic dysregulation and the overall profile lent support to the transcriptomics data that temporal effects had a greater influence on the changes observed. Exposure to handling-confinement stress revealed that ozone treatment did not alter the ability of post-smolts to respond to a secondary stressor. In summary, the suite of health and welfare indicators collectively indicated that continuous ozonation resulted in minimal physiological perturbations in salmon post-smolts. The results are expected to contribute to optimising the rearing conditions for post-smolts in RAS.

Effect of topical ozonated sunflower oil on second intention wound healing in turtles: a randomised experimental study.

BackgroundOzone is an antimicrobial agent that in experimental and case-control studies has been found to exert a positive effect on wound healing. Wild and pet chelonians frequently present insidious wounds exhibiting secondary infections and/or delayed healing.ObjectivesEvaluate the effects of topical ozonated sunflower oil on second-intention healing of acute experimental skin wounds in red-eared sliders (Trachemys scripta elegans).MethodsRandomised within-subject controlled study; Group 1 (n = 24) was used to assess clinical healing features; Group 2 (n = 12) was used for histological evaluation in which two sets of wounds were biopsied at 2, 7, 14, 21, 28 and 42 days over the course of the cicatrisation process. A single 6 mm diameter wound was made on each rear limb and topical ozonated (950 peroxide value) and non-ozonated sunflower oil were applied daily for one week on treated and contralateral control wounds, respectively.ResultsMean wound size was significantly lower in the ozone-treated group at day 28 (p < 0.0001) with differences of clinical relevance (74.04% vs. 93.05% reduction of initial wound size). Histologically, the acute inflammatory reaction was enhanced in treated wounds, with significantly higher numbers of heterophils (p = 0.0016), lymphocytes (p < 0.001) and fibroblasts (p < 0.001).ConclusionsDaily topical application of ozonated sunflower oil over the course of one week improved the healing of acute, full-thickness skin wounds in chelonians. This clinical outcome was histologically correlated with an enhanced acute inflammatory reaction, as well as the production and remodelling of collagen fibres.

The effects of ozone therapy on postoperative adhesions and ovarian functions: An experimental study

Aim: Numerous methods are used to prevent the development of postoperative adhesion formation. There are few studies on the effects of ozone therapy on postoperative intraabdominal adhesions and ovarian functions. This study aimed to investigate the effects of ozone treatment on postoperative intraabdominal adhesions and ovarian roles in rats.Methods: Twenty female Wistar albino rats were randomly allocated into two groups as laparotomy (Group 1, n: 10) and laparotomy + intraperitoneal ozone (Group 2, n: 10). After laparotomy, parietal peritoneum, uterus and adnexal surfaces were scraped in both groups. Follicle-stimulating hormone (FSH), Estradiol (E2), Lactate Dehydrogenase (LDH), Urea, and Creatinine levels were measured, and histopathological evaluation was performed. Both groups were compared concerning histopathological and biochemical findings.Results: In the ozone-treated group, antioxidant levels (P=0.012) were significantly higher, and E2 levels (P=0.005) were substantially lower than the control group. There was no statistically significant difference between the two groups regarding FSH, Urea, Creatinine, and LDH (P=0.12, P=0.72, P=0.45, and P=0.79, respectively). Histopathologically, postoperative intraabdominal adhesion rates between the two groups were statistically similar.Conclusion: Although there was no statistically significant difference, ozone therapy had a decreasing effect on severe fibrosis and congestion rates. Although there was no difference in FSH, low levels of E2 in Group 2 suggest that ozone treatment may have a protective effect on the ovaries. However, further studies are needed concerning adhesion formation and the impact on ovarian functions.

Efficacy of ozonized water for the treatment of erosive oral lichen planus: a randomized controlled study.

Background Management of erosive Oral Lichen Planus (eOLP) is challenging. Currently, topical corticosteroids are widely used as first-line therapy, but they might be associated with side-effects and incomplete clinical response. Among non-pharmacological strategies, ozone at low medical concentration has proven to induce a mild activation of protective anti-oxidant pathways, thus exerting therapeutic effects in many inflammatory diseases. The aim of this randomized controlled study was to investigate the effectiveness of ozonized water in association with conventional topical corticosteroids for the treatment of eOLP. Material and Methods Fifty-one patients were included in the study and randomized into 2 groups: study group (n=26) included patients receiving ozonized water treatment; control group (n=25) included patients receiving placebo treatment (i.e. double-distilled water). Treatment protocol consisted of 1-minute oral rinses, repeated for 4 times, twice a week for 4 weeks. All patients received conventional corticosteroid topical therapy (betamethasone soluble tablets, 2 rinses/day for 4 weeks). Assessment of size of lesions, sign and pain scores was performed before treatment, after 2 weeks of treatment (T1) and at the end of 4-week treatment (T2). Efficacy Index (EI) of treatment, candidiasis and relapse rates were also recorded. Results All patients experienced significant improvement of sign and pain scores with a higher rate of improvement in ozone-treated group (T1 improvement rates: Thongprasom 92.2% vs 28%; VAS pain 76.9% vs 32%; p<0.05). Pain and size reduction were significantly higher in ozone-treated group both at T1 and T2 (p<0.05). Ozone-treated group showed a higher EI at every time point (T0-T2: 72.77% vs 37.66%, p<0.01). Candidiasis (32% vs 11.5%) and relapse (40% vs 34.6%) rates were higher in control group, however the differences were not statistically significant. Conclusions Within the limitations of this study, ozonized water seems to be effective as an adjunct therapy, in combination with topical corticosteroids, for the treatment of eOLP. Key words:Oral Lichen Planus, OLP, ozone, treatment.

Enhancing bone healing in calvarial critical size defect using Ozone gel: histological and Histomorphometric analysis.

The aim of this study was to examine the validity of the hypothesis that Ozone gel will accelerate bone healing in a critical size defect model experimentally created in rabbit calvaria when mixed with autografts in comparison to autogenous grafts per se. Materials and Methods: A total of twelve adult male New Zealand Rabbits were included in the study. A total of 24 grafts were harvested from 12 animals, each graft was crushed using a special bone mill device. After bone milling, each bone graft was collected in a special sterile container, twelve grafts were mixed with the normal saline solution (control group) and each one of the rest of the grafts was mixed with ozone gel. Animals were sacrificed at 7 and 14 days’ post-surgery, dual energy x-ray absorptiometry (DEXA) scans were performed for the skull of the rabbits and bone specimens were collected for histological examination. Results: Histomorphometric analysis showed superior results in favor of the ozone-treated group represented as a significantly higher percentage of normal osteocytes and marked increase in area percentage of the new bone formation. Additionally, DEXA scan revealed a significant increase in bone mineral density and bone mineral concentration of the ozone-treated group compared to the control group. Conclusion: The use of ozone gel may be cost-effective and convenient owing to its ease of preparation. It is recommended to be used with routine bone grafting procedures.

Effects of Ozone Therapy on the Early Healing Period of Deepithelialized Gingival Grafts: A Randomized Placebo-Controlled Clinical Trial.

This study evaluates the effects of ozone therapy (OT) on the early healing period of deepithelialized gingival grafts (DGG) placed for non-root coverage gingival augmentation by laser Doppler flowmetry (LDF). Thirty-three patients were assigned to study groups: 1) test: DGG + OT; or 2) control group: DGG alone. Thirty patients completed the study. Ozone was applied on DGGs placed in the recipient bed and donor site immediately after surgery and at days 1 and 3 post-surgery. Blood perfusion in the recipient site was measured by LDF on the day of surgery and at 1, 2, 3, 6, 8, 10, and 13 days after surgery. Quality of life (assessed by the Oral Health Impact Profile-14) and pain at donor/recipient sites (assessed by visual analog scale) were also investigated. Increase in blood perfusion units in the test group was significantly higher than control group at 1, 2, 3, 6, and 8 days post-surgery (P <0.001). Significant differences occurred between test and control groups in terms of visual analog scale values during the first week post-surgery for both donor and recipient sites (P <0.05). The ozone-treated group showed significantly higher quality of life than control group on postoperative day 6 (P = 0.002). OT enhanced blood perfusion units in the first postoperative week. This outcome is also consistent with improvement in wound healing, accompanied by an increase in quality of life and decrease in postoperative pain in the test group.

Effects of ozone therapy on facial nerve regeneration

IntroductionOzone may promote moderate oxidative stress, which increases antioxidant endogenous systems. There are a number of antioxidants that have been investigated therapeutically for improving peripheral nerve regeneration. However, no previous studies have reported the effect of ozone therapy on facial nerve regeneration. ObjectiveWe aimed to evaluate the effect of ozone therapy on facial nerve regeneration. MethodsFourteen Wistar albino rats were randomly divided into two groups with experimental nerve crush injuries: a control group, which received saline treatment post-crush, and an experimental group, which received ozone treatment. All animals underwent surgery in which the left facial nerve was exposed and crushed. Treatment with saline or ozone began on the day of the nerve crush. Left facial nerve stimulation thresholds were measured before crush, immediately after crush, and after 30 days. After measuring nerve stimulation thresholds at 30 days post-injury, the crushed facial nerve was excised. All specimens were studied using light and electron microscopy. ResultsPost-crushing, the ozone-treated group had lower stimulation thresholds than the saline group. Although this did not achieve statistical significance, it is indicative of greater functional improvement in the ozone group. Significant differences were found in vascular congestion, macrovacuolization, and myelin thickness between the ozone and control groups. Significant differences were also found in axonal degeneration and myelin ultrastructure between the two groups. ConclusionWe found that ozone therapy exerted beneficial effect on the regeneration of crushed facial nerves in rats.

EFFECT OF OZONE THERAPY ON SPERMATOGENESIS IN EXPERIMENTALLY GENTAMYCIN-INDUCED TESTICULAR LESION IN ADULT MALE ALBINO RAT

Background: Spermatogenesis is a complex process through which spermatogonial germ cells undergo mitosis, meiosis and cell differentiation to generate mature spermatozoa. Gentamycin (GM) is an aminoglycoside antibiotic that has negative effect on spermatogenesis. Objective: Evaluation of the possible effect of medical ozone on spermatogenesis in experimentally gentamycin-induced testicular lesion in male albino rat. Materials and Methods: Forty adult male albino rats of a local strain were chosen as a model for the present study. They were divided into equal four groups: Group I (Control group), Group II (Gentamycin-treated group), GroupIII (Ozone and gentamycin-treated group), and Group IV (Gentamycin followed by ozone-treated group). At the end of the experimental period, blood samples were obtained for determination of malondilaldehyde (MDA) and catalase (CAT). Testicular samples were obtained for transmission electronmicorsocpic study. Results: Gentamycin induced oxidative stress and reduced spermatogenesis. Medical ozone produced partial improvement in redox homeostasis and spermatogenesis when used in combination or after gentamycin. Conclusion: Medical ozone can be used to attenuate harmful effects of GM on testicular functions. Further studies can be done to evaluate the effect of different doses and different durations of medical ozone on different systems of the body.

Effect of ozone to remineralize initial enamel caries: in situ study.

Effect of ozonated water in remineralizing artificially created initial enamel caries was investigated using laser fluorescence and polarized light microscopy in an in situ study. Teeth specimens (buccal sections) were immersed in 5-ml solution of 2mM CaCl2, 2mM NaH2P04, and 50mM CH3COOH at pH of 4.55 for 5h in an incubator at 37° to create subsurface demineralization. After which, they were randomly allocated into one of the following remineralization regimens: ozone (ozonated water 0.1mg/l and 10% nano-hydroxyapatite paste, Aclaim(TM)), without ozone (only 10% nano-hydroxyapatite paste, Aclaim(TM)), and control (subjects' saliva alone). Specimens were embedded in acrylic retainers worn by orthodontic patients throughout the 21-day study duration and constantly exposed to their saliva. Laser fluorescence was recorded for all the specimens at baseline, after demineralization, and remineralization using DIAGNOdent, and the results were validated using polarized microscopic examination. The results were analyzed using repeated measures, one-way ANOVA with post hoc multiple comparisons. Reduced DIAGNOdent scores and greater depth of remineralization following application of ozonated water and nano-hydroxyapatite were found compared to those of the without ozone and control groups (P < 0.001), and the ozone-treated group exhibited maximum remineralization under the polarized light microscopy. Ozonated water can be considered an effective agent in reversing the initial enamel caries alongside with nano-hydroxyapatite compared to nano-hydroxyapatite alone and saliva. Ozone water can be used to remineralize incipient carious lesions, and it enhances the remineralizing potential of nano-hydroxyapatite thereby preventing the tooth from entering into the repetitive restorative cycle.

The Role of Ozone Therapy in Hepatic Fibrosis due to Biliary Tract Obstruction.

In this experimental study, we investigated the role of ozone therapy in hepatic fibrosis caused by biliary obstruction. In this study, 21 male Sprague-Dawley rats were divided equally into three groups. In the control group, only laparotomy was performed and intraperitoneal cavity is washed with 1 mL of saline. In the sham group (SG), common bile duct is binded with laparotomy and no treatment is given afterward. In the experimental group (EG) after the binding of common bile duct with laparotomy, 1 mg/kg dose and 50 mg/mL concentration of ozone were applied rectally for 21 days. Hepatic tissue sample and intracardiac blood samples were collected from all animals at postoperative 21st day with relaparatomy. When we compared the experiment to SG, we detected a decrease in aspartate aminotransferase, alkaline phosphate (ALP), total bilirubin, and direct bilirubin levels in the EG, however, only the decrease in total bilirubin levels were statistically significant (p = 0.025). Histopathological examination of livers of rats in the EG showed lower level of hepatic fibrosis and inflammation. In the SG, incomplete cirrhosis was detected in 57.1% of the rats, whereas in the EG, no cirrhosis was detected. Immunohistochemically, periportal inflammation was 100% in the SG, whereas it was seen (3/7) 42.9% in the EG. A significant decrease in positive α-smooth muscle actin reaction was observed in ozone-treated group compared with SG. We suggest that ozone can decrease the hepatic destruction levels in experimental model of biliary obstruction.

EFFECT OF OZONE THERAPY ON ISCHEMIC REPERFUSION INJURY OF THE HEART AND VASCULAR REACTIVITY IN ADULT MALE DIABETIC ALBINO RAT

Background: Ozone therapy is a form of complementary medicine treatment that aims to increase the amount of oxygen to the body through the introduction of ozone into the body. Objective: Studying the effects of diabetes mellitus, insulin and ozone on ischemic reperfusion (IR) injury on the heart and vascular reactivity in diabetic rat. Material and Methods: Two hundred and fifty adult male albino rats of local strain, weighing 120-150 ±10 grams each, were used in this investigation and divided into: Group I (Normal control group): 10 rats. Group II (Diabetic group -240 rats) were subdivided into 4 subgroups: Group II-a (Diabetic non-treated group - 40 rats), Group II-b (Diabetic insulin-treated group - 40 rats, and Group II-c (Diabetic ozone-treated group - 80 rats Group II-d (Diabetic ozone and insulin-treated group - 80 rats): Rats were submitted to ozone therapy with concomitant treatment with insulin. Supernatant serum was collected in a dry clean tube for estimation of fasting serum glucose, serum total cholesterol, triglycerides, HDL, LDL, LDH, catalase enzyme, glutathione peroxidase, and SOD. Rat aortic rings preparation were used for estimation of changes in vascular reactivity in response to norepinephrine (10-5), vasopressin (10-6 M), indomethacin (10-6 M) and relaxation of aortic rings (preconstricted by NE (10-5) in response to ACh (10-6) and Na+ nitroprusside (10-6) as estimated in different groups. The organ bath was washed out three times with fresh Krebs' solution before the next substance was added and the rings were allowed to stabilize for 1 hour. All results were presented as the mean ± SEM. The data were analyzed using SPSS program version 12. For comparison of statistical significance between different groups, a one way ANOVA with the post hoc of Tukey's multiple comparison test was used. A value of P ≤ 0.05 was considered statistically significant. Results: Ozone therapy caused significant decrease in fasting glucose, total cholesterol, triglycerides, LDL, and lactate dehydrogenase “LDH”, and significant increase in HDL and myocardial antioxidants (catalase, superoxide dismutase “SOD” and glutathione peroxidase). There were a significant increase in cardiac contractility and heart rate during pre-ischemic and ischemic periods. There was a significant decrease in heart rate accompanied by significant increase in cardiac contractility during reperfusion period. Also, ozone treatment produced a significant decrease in vascular reactivity of aortic rings to norepinephrine, vasopressin and indomethacin, with significant increase in percentage of relaxation to acetyl choline “ACh”. Conclusion: Diabetic complications are attributed to the oxidative stress in the body. Ozone activates the antioxidant system affecting the level of glycemia. Ozone prevents oxidative stress by normalizing the organic peroxide levels by activating superoxide dismutase.