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Background: The development of periodontal disease has been thought to be associated with several restricted members of the oral anaerobic species, such as black-pigmented Porphyromonas species and Actinobacillus actinomycetemcomitans (Aa), in the subgingival environment. Apart from bacteria, certain viruses and fungi that are associated with periodontal disease are also present in the subgingival plaque. Materials and Methods: A randomized, double-blind, crossover split-mouth design was performed. A total of 16 patients suffering from generalized chronic periodontitis were selected for the study. The study period of 18 days was divided into two time-intervals, i.e. baseline (0 days) to 7 th day, with a washout period of 4 days followed by a second time interval of 7 days. The use of ozone and chlorhexidine gluconate (CHX) irrigation was randomized. Both the patient and the clinician evaluating the clinical parameters were blinded regarding the type of irrigation used. Results: The interpretation of clinical and microbial data is from baseline to 7 th day. A higher percentage of plaque index (12%), gingival index (29%) and bleeding index (26%) reduction was observed using ozone irrigation as compared to chlorhexidine. The percentile reduction of Aa (25%) using ozone was appreciable as compared to no change in Aa occurrence using chlorhexidine. By using O 3 and chlorhexidine, there was no antibacterial effect on Porphyromonas gingivalis (Pg) and Tannerella forsythensis. The antifungal effect of ozone from baseline (37%) to 7 th day (12.5%) was pronounced during the study period, unlike CHX, which did not demonstrate any antifungal effect. Conclusion: Ozone may be considered as an alternative management strategy due to its powerful ability to inactivate microorganisms. Also, there is growing evidence that ozone can be employed as a useful therapeutic agent in both dentistry and medicine.

Objective: The purpose of this study was to investigate the remineralization activation of the application of three fissure sealants (FSs), alone or with gaseous ozone (GO), on non-cavitated initial caries and evaluate the clinical success of FS. Study design: Sixty children who had DIAGNOdent scores between 10–30 on bilateral symmetric mandibular first permanent molars were included in study. In a split-mouth design, teeth were assigned to experimental (with GO) and control (without GO) groups. GO was applied to teeth on one side and then the same brand of randomly selected FSs was applied to the teeth on both sides. Children were divided into 3 groups based on type of FS (Group 1: Aegis Bosworth Co, North Hamlin Avenue Skokie, Illinois, USA, Group 2: Fuji Triage GC, Tokyo, Japan, Group 3: Helioseal Ivoclar Vivadent, Liechtenstein, Germany). All FSs were then examined for retention rates at 1, 3, 6, 9, and 12 months; at the end of 12 months, all FSs were removed with an air-abrasion device and DIAGNOdent scores noted to compare with the initial values. Results: The application of GO with either Fuji Triage or Aegis FS was effective on remineralization (p<0.05); however, the application of Helioseal FSs was not effective (p>0.05). The 1st and 12th months’ full retention rates of Fuji Triage FSs was a significant difference (p<0.05) from other FSs (Aegis and Helioseal) (p>0.05). Conclusions: GO+Aegis FS showed the highest remineralization; and, at the end of 12 months, its clinical success was higher than other FSs.

The ability of ozone to kill micro-organisms associated with non-cavitated occlusal caries was investigated. The occlusal surfaces were treated with ozone (n = 53) or air (n = 49) for 40 s, and the underlying infected dentine was exposed. There was no significant difference between the number of bacteria recovered from the ozone-treated and the control sites (p > 0.1). Treatment of the exposed dentine with ozone resulted in a just significant (p = 0.044) reduction in bacterial counts. Ozone treatment of non-cavitated occlusal lesions for 40 s failed to significantly reduce the numbers of viable bacteria in infected dentine beneath the demineralized enamel.

Background This study evaluates the effects of ozone on hard and soft tissue healing when a free tissue flap is used to close wound areas lacking primary closure over autogenous grafted sites. Methods In our study, 24 male Wistar rats were divided into four groups: two control groups and two ozone-treated groups. All rats underwent the same surgical procedure. After surgery, the control groups received no additional treatment, while the ozone groups received topical ozone application. Ozone was applied for 2 min, three times per week, with one-day intervals, for a duration of 2 weeks post-surgery. Sacrifice was performed on the 14th day for one control and one ozone group, and on the 28th day for the remaining groups. All soft and hard tissue samples were then examined histopathologically. Clinical trial number: not applicable. Results Histopathological evaluations and statistical analyses on the 14th day indicated that bone healing was significantly better in the ozone group compared to the control group ( p  = 0.049). For soft tissue healing, although there was no statistically significant difference between the ozone and control groups, proportional values showed better recovery in the ozone-treated groups compared to the controls. Conclusions The present study concludes that ozone has a positive effect on new bone formation in the early stages; however, it showed no statistically significant effect on soft tissue healing. We believe that further comprehensive studies would be beneficial to confirm these findings.

Background Accelerated apoptosis plays a vital role in the development of diabetic vascular complications. Ozone may attenuate diabetic nephropathy by means of decreased apoptosis-related genes. The aim of our study was to investigate the effect of ozone therapy on streptozotocin-induced diabetic nephropathy in rats. Also the histopathological changes in diabetic kidney tissue with ozone treatment were evaluated. Methods The rats were randomly divided into six groups ( n  = 7): control (C), ozone (O), diabetic (D), ozone-treated diabetic (DO), insulin-treated diabetic (DI), and ozone- and insulin-treated diabetic (DOI). D, DI, and DOI groups were induced by a single intraperitoneal injection of streptozotocin. Ozone was given to the O, DO, and DOI groups. Group DI and DOI received subcutaneous (SC) insulin (3 IU). All animals received daily treatment for 6 weeks. Results Expressions of caspase-1-3-9, HIF-1α, and TNF-α genes were significantly higher in D group compared to C group ( p  < 0.05 for all). Ozone treatment resulted in significant decrease in the expressions of these genes in diabetic kidney tissue compared to both C and D group ( p  < 0.05 for all). Caspase-1-3-9, HIF-1α, and TNF-α gene expressions were found to be lower in DOI group compared to C group ( p  < 0.05 for all). Also adding ozone treatment to insulin therapy resulted in more significantly decrease in the expressions of these genes in diabetic tissue compared to only insulin-treated diabetic group ( p  < 0.05 for all). Regarding histological changes, ozone treatment resulted in decrease in the renal corpuscular inflammation and normal kidney morphology was observed. Both insulin and ozone therapies apparently improved kidney histological findings with less degenerated tubules and less inflammation of renal corpuscle compared to D, DO, and DI groups. Conclusion Ozone therapy decreases the expressions of apoptotic genes in diabetic kidney tissue and improves the histopathological changes.

Medical ozone has therapeutic properties as an antimicrobial, anti-inflammatory, modulator of antioxidant defense system. Major ozonated autohemotherapy (MOA) is a new therapeutic approach that is widely used in the treatment of many diseases. The objective of the present study was to determine whether preischemic application of MOA would attenuate renal ischemia-reperfusion injury (IRI) in rabbits. Twenty-four male New Zealand white rabbits were divided into four groups, each including six animals: (1) Sham-operated group, (2) Ozone group (the MOA group without IRI), (3) IR group (60 min ischemia followed by 24 h reperfusion), and (4) IR + MOA group (MOA group). The effects of MOA were examined by use of hematologic and biochemical parameters consisting of neutrophil to lymphocyte ratio (NLR), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), ischemia-modified albumin (IMA), total antioxidant status (TAS), total oxidant status (TOS), and oxidative stress index (OSI). In addition, the histopathological changes including the tubular brush border loss (TBBL), tubular cast (TC), tubular necrosis (TN), intertubular hemorrhage and congestion (IHC), dilatation of bowman space (DBS), and interstitial inflammatory cells infiltration (IECI) were evaluated. In the IR group, compared to the Sham group, biochemical parameters indicating oxidative stress, NLR, IL-6, TNF-α, IMA, TOS, and OSI have increased. MOA reduced inflammation and oxidative stress parameters. Although TAS values have decreased in the IR group and increased in the MOA-pretreated group, no significant changes in TAS values were detected between the IR and MOA groups. The total score was obtained by summing all the scores from morphological kidney damage markers. The total score has increased with IR damage when compared with the Sham group (13.83 ± 4.30 vs 1.51 ± 1.71; p  = 0.002). But, the total score has decreased significantly after application of MOA (5.01 ± 1.49; p  = 0.002; compared with the IR group). MOA preconditioning is effective in reducing tissue damage induced in kidney ischemia-reperfusion injury. The protective effect of MOA is mediated via reducing inflammatory response and regulating of reactive oxygen species (ROS). Renal histology also showed convincing evidence regarding MOA’s protective nature against kidney injury induced renal ischemia-reperfusion. Consequently, MOA might be helpful in protecting the kidneys from IR-induced damage in humans, probably through the anti-inflammatory effect and reducing the total oxidant status.

Because radiation-induced cellular damage is attributed primarily to harmful effects of free radicals, molecules with direct free radical scavenging properties are particularly promising as radioprotectors. It has been demonstrated that controlled ozone administration may promote an adaptation to oxidative stress, preventing the damage induced by reactive oxygen species. Thus, we hypothesized that ozone would ameliorate oxidative damage caused by total body irradiation (TBI) with a single dose of 6 Gy in rat liver and ileum tissues. Rats were randomly divided into groups as follows: control group; saline-treated and irradiated (IR) groups; and ozone oxidative preconditioning (OOP) and IR groups. Animals were exposed to TBI after a 5-day intraperitoneal pretreatment with either saline or ozone (1 mg/kg/day). They were decapitated at either 6 h or 72 h after TBI. Plasma, liver and ileum samples were obtained. Serum AST, ALT and TNF-α levels were elevated in the IR groups compared with the control group and were decreased after treatment with OOP. TBI resulted in a significant increase in the levels of MDA in the liver and ileal tissues and a decrease of SOD activities. The results demonstrated that the levels of MDA liver and ileal tissues in irradiated rats that were pretreated with ozone were significantly decreased, while SOD activities were significantly increased. OOP reversed all histopathological alterations induced by irradiation. In conclusion, data obtained from this study indicated that ozone could increase the endogenous antioxidant defense mechanism in rats and there by protect the animals from radiation-induced organ toxicity.

The aims of this present study were (1) to assess the antimicrobial effect of ozone from a novel ozone–generating device (Heolozone, USA) [0.052% (v/v) in air delivered at a rate of 13.33 ml·s –1 ] on primary root carious lesions (PRCLs) and (2) to evaluate the efficacy of ozone specifically on Streptococcus mutans and Streptococcus sobrinus. In study 1, 40 soft PRCLs from freshly extracted teeth were randomly divided into two groups to test the antimicrobial effect on PRCLs from exposure to ozonated water for either 10 or 20 s. Half of a lesion was removed using a sterile excavator. Subsequently, the remaining lesion was exposed to the ozonised water for a period of either 10 or 20 s (corresponding to 0.069 or 0.138 ml of ozone, respectively). Using paired Student t tests, a significant (p<0.001) reduction (mean ± SE) was observed in the ozone–treated groups with either a 10–second (log 10 3.57±0.37) or 20–second (log 10 3.77±0.42) ozone application compared with the control groups (log 10 5.91±0.15 and log 10 6.18±0.21, respectively). In study 2, 40 sterile saliva–coated glass beads were randomly divided into two groups for each micro–organism. One glass bead was put into each bijou bottle with 3 ml of Todd–Hewitt broth. S. mutans and S. sobrinus were inoculated anaerobically overnight. Each glass bead was then washed with 2 ml of phosphate–buffered saline. Immediately, 10 s of ozone gas was applied to each glass bead in the test groups. There was a significant (p<0.0001) reduction (mean ± SE) in ozone–treated samples for S. mutans (log 10 1.01±0.27) and S. sobrinus (log 10 1.09±0.36) compared with the control samples (log 10 3.93±0.07 and log 10 4.61±0.13, respectively). This treatment regime is an effective, quick, conservative and simple method to kill micro–organisms in PRCLs. Ozone gas application for a period of 10 s was also capable of reducing the numbers of S. mutans and S. sobrinus on saliva–coated glass beads in vitro.

PurposeVarious studies have been performed to find out novel treatment strategies to prevent postoperative adhesion formation. Ozone therapy (OT) is shown to reduce inflammation in several pathological conditions. Therefore, we aimed to evaluate the efficacy of OT in a rat model of experimental uterine adhesion (EUA).MethodsThirty female Wistar rats (200–250 g) were divided into three groups: sham, EUA and EUA+OT. EUA and EUA+OT groups were subjected to the postoperative adhesion procedure by bipolar coagulation on the uterine horns and corresponding pelvic sidewall parietal peritoneum. EUA+OT group received 0.7 mg/kg daily single dose for 3 days of ozone/oxygen mixture intraperitoneally after adhesion induction. All animals were killed on the 7th day and uterine adhesions were scored. Uterine tissues and peritoneal washing fluid were harvested for all analyses.ResultsUterine malondialdehyde levels in the EUA group were significantly higher compared to the other groups. However, in the EUA group, uterine superoxide dismutase and glutathione peroxidase activities were lower than in other groups. Peritoneal fluid TNF-α levels were found to be significantly different for all groups (p < 0.001). Macroscopic total adhesion score was significantly higher in the EUA group compared to the other groups (p < 0.001). But, total score in the EUA+OT group was lower than in the EUA group (p = 0.006).ConclusionsMedical OT prevents postoperative uterine adhesions by modulating TNF-α levels and oxidative/antioxidative status in an experimental uterine adhesion model.

The confirmed advantageous effects of oxygen/ozone therapy in several clinical conditions stimulated experimental studies on effects of the therapy in rats with an induced septic shock. The studies were conducted on adult male rats of Wistar strain. Four groups of the animals, each of 15 rats, included: I--control group, (C); II--animals intraperitoneally administered with O(2)/O(3) (CO), III--rats given of Escherichia coli endotoxin (lipopolysaccharide-LPS) (CL), IV--rats administered with the lipopolysaccharide plus administered with the oxygen/ozone mixture (OL). Activities of catalase and superoxide dismutase and of free radical reactions were estimated. The exposure to LPS augmented activities of SOD and of catalase in liver, lungs and heart. In all the examined organs LPS induced significant changes in levels of free radicals. Except of the lungs, parallel administration of the rats with LPS and ozone/oxygen revoked development of the alterations. The obtained results point to a strong, stabilizing and regenerative effect of ozonotherapy.