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Ozone in Dentistry ( II )

See Also : Ozone Therapy ( Index )  //  Ozone In Dentistry ( I )

http://www.thesleuthjournal.com
http://www.newagegathering.com/ozonated-olive-oil-promotes-oral-health/

Ozonated Olive Oil Promotes Oral Health
by Dr. Edward F. Group III

The use of oxygen in complementary therapies has been gaining momentum in recent years, particularly in the area of dentistry. While ozonated olive oil has many practical and effective uses, such as supporting skin health, research has shown it to be especially helpful for periodontal disease. Gingivitis, typically caused by diets laden with sugar and refined carbohydrates, is a growing issue today. One recent study is revealing just how powerful ozonated therapy can be in the area of teeth and gum health. If you’re currently suffering from any type of dental issue, you may want to continue reading.

Olive Oil: A Potential Complementary Periodontitis Approach

Ozone therapy isn’t an entirely new concept in dentistry. Previous research and medical reviews have discussed the importance of ozone in replacing antiseptic agents used against gingival infections. [1] A study published in Tanta Dental Journal evaluated 30 subjects with periodontal disease, each receiving standard periodontitis treatments. [2] Half of the group was subjected to both standard treatments and ozonated therapy. These patients received ozonated olive oil gel applied to the main site of gum damage.

The group that received the complementary therapy in addition to conventional treatment saw longer lasting improvements in symptoms. As a matter of fact, the ozonated therapy combined with root scaling resulted in sustained improvements for up to six months, while the benefits from the standard treatment alone only lasted about a month. Although the size of the study participants is relatively small, it is one study among dozens that indicate the effectiveness of ozone therapy in complementing standard medical applications.

Using Ozonated Olive Oil

One of the main uses for ozonated olive oil is in the area of skin health. Ozonated olive oil contains antibacterial compounds that may support the skin’s defense against bacteria and other compounds that cause acne. Olive oil itself is also an excellent moisturizer, providing a nourishing smoothness to the skin. Many people also brush their teeth with ozonated olive oil in addition to fluoride-free toothpaste to get a deep clean.

References:

Huth KC, Jakob FM, Saugel B, et al. Effect of ozone on oral cells compared with established antimicrobials. Eur J Oral Sci. 2006 Oct;114(5):435.

M.Y.M. Shoukheba, Sh.A. Ali. The effects of subgingival application of ozonated olive oil get in patient with localized aggressive periodontitis. A clinical and bacteriological study. Tanta Dental Journal. Volume 11, Issue 1, April 2014, Pages 63-73.

Dr. Edward F. Group III, DC, NP, DACBN, DCBCN, DABFM has studied natural healing methods for over 20 years and now teaches individuals and practitioners all around the world. He no longer sees patients but solely concentrates on spreading the word of health and wellness to the global community. Under his leadership, Global Healing Center, Inc. has earned recognition as one of the largest alternative, natural and organic health resources on the Internet.



http://www.ncbi.nlm.nih.gov/pubmed/17026511

Eur J Oral Sci. 2006 Oct;114(5):435-40.

Effect of ozone on oral cells compared with established antimicrobials.

Huth KC, Jakob FM, Saugel B, Cappello C, Paschos E, Hollweck R, Hickel R, Brand K.

Abstract

Ozone has been proposed as an alternative antiseptic agent in dentistry based on reports of its antimicrobial effects in both gaseous and aqueous forms. This study investigated whether gaseous ozone (4 x 10(6) microg m(-3)) and aqueous ozone (1.25-20 microg ml(-1)) exert any cytotoxic effects on human oral epithelial (BHY) cells and gingival fibroblast (HGF-1) cells compared with established antiseptics [chlorhexidine digluconate (CHX) 2%, 0.2%; sodium hypochlorite (NaOCl) 5.25%, 2.25%; hydrogen peroxide (H(2)O(2)) 3%], over a time of 1 min, and compared with the antibiotic, metronidazole, over 24 h. Cell counts, metabolic activity, Sp-1 binding, actin levels, and apoptosis were evaluated. Ozone gas was found to have toxic effects on both cell types. Essentially no cytotoxic signs were observed for aqueous ozone. CHX (2%, 0.2%) was highly toxic to BHY cells, and slightly (2%) and non-toxic (0.2%) to HGF-1 cells. NaOCl and H(2)O(2) resulted in markedly reduced cell viability (BHY, HGF-1), whereas metronidazole displayed mild toxicity only to BHY cells. Taken together, aqueous ozone revealed the highest level of biocompatibility of the tested antiseptics.



http://www.ncbi.nlm.nih.gov/pubmed/25512726
Eur J Dent. 2014 Oct;8(4):469-74. doi: 10.4103/1305-7456.143627.

Antimicrobial effect of ozonated water, sodium hypochlorite and chlorhexidine gluconate in primary molar root canals.

Goztas Z, Onat H, Tosun G, Sener Y, Hadimli HH.

Abstract

OBJECTIVE:


The aim was to determine the antimicrobial effect of ozonated water, ozonated water with ultrasonication, sodium hypochloride and chlorhexidine (CHX) in human primary root canals contaminated by Enterococcus faecalis (E. faecalis).

MATERIALS AND METHODS:

Fifty-eight extracted human primary molar teeth were used. Crowns were cut off using a diamond saw under water-cooling. One hundred roots were obtained and mechanically prepared. The roots were then sterilized by autoclaving in water for 15 min at 121°C. All samples were contaminated with E. faecalis for 24 h and the root canals were randomly divided into five groups (n = 20). Group I: 25 mg/L of Ozonated water (O3aq), Group II: 25 mg/L of O3aq with ultrasonication, Group III: 2.5% Sodium hypochloride (NaOCl), Group IV: 2% CHX and Group V: Positive control. The canal of each specimen was irrigated for 4 min and positive control was untreated. All root canals were agitated with sterile saline solution. The saline solution was collected from canals with sterile paper points. For each specimen, the paper points were transposed to eppendorf vials containing 2 ml of brain heart infusion. According to bacterial proliferation, the mean values of optical density were achieved by ELISA (Biotek EL ×800, Absorbance Microplate Reader, ABD) and the data were analyzed.

RESULTS:

NaOCI, CHX and two types of O3aq were found statistically different than positive control group. NaOCI irrigation was found significantly most effective.

CONCLUSIONS:

NaOCl, CHX and O3aq applications provide antibacterial effect in vitro conditions in primary root canals.



http://www.ncbi.nlm.nih.gov/pubmed/25368798
J Microbiol. 2014 Jul;7(7):e11411. doi: 10.5812/jjm.11411. Epub 2014 Jul 1.

Antibacterial Efficacy of Aqueous Ozone in Root Canals Infected by Enterococcus faecalis.

Hubbezoglu I, Zan R, Tunc T, Sumer Z.

Abstract

BACKGROUND:

In endodontics, the elimination of resistant bacteria such as Enterococcus faecalis plays an important role for treatment success in root canals. Therefore, new alternative irrigants (instead of sodium hypochlorite) have been researched to achieve ideal endodontic treatment.

OBJECTIVES:

The aim of the present study was to evaluate and to compare the antibacterial effect of aqueous ozone with different concentrations and techniques of application (manual and ultrasonic) against E. faecalis in human root canals.

PATIENTS AND METHODS:

Eighty single-root mandibular premolar teeth were selected, prepared and sterilized. E. faecalis was incubated in the root canals and kept at 37°C for 24 h. The teeth were divided into four main groups each has 20 members: NaOCl (positive control) group; 8 ppm aqueous ozone group; 12 ppm aqueous ozone group; and 16 ppm aqueous ozone group. While half of the specimens were disinfected with aqueous ozone by manual technique, the other half was disinfected with the aqueous ozone by ultrasonic technique. Conventional irrigation technique was simultaneously applied with ultrasonic vibration that was produced by VDW.ULTRA device. The disinfection procedures were performed for 180 s to ensure standardization of all the working groups. Paper points (placed in the root canals before and after the disinfection procedures) were transferred to Eppendorf tubes containing 0.5 mL of brain heart infusion broth. Then, 50 µL of the suspension was inoculated onto broth agar media. Microbial colonies were counted, and the data were evaluated statistically using 2-way analysis of variance (ANOVA) and Tukey tests.

RESULTS:

Although the antibacterial effect of 16 ppm aqueous ozone using a manual technique had an insufficient effect, its ultrasonic application technique resulted in complete disinfection in the root canals.

CONCLUSIONS:

The bactericidal activity of high concentration of aqueous ozone combined with ultrasonic application technique showed efficacy similar to that of 5.25% NaOCl in root canals.



http://www.ncbi.nlm.nih.gov/pubmed/25363268
Interv Neuroradiol. 2014 Oct 31;20(5):632-6. doi: 10.15274/INR-2014-10083. Epub 2014 Oct 17.

Ozone therapy in dentistry. A brief review for physicians.

Domb WC

Abstract

The 21(st) century dental practice is quite dynamic. New treatment protocols and new materials are being developed at a rapid pace. Ozone dental therapy falls into the category of new treatment protocols in dentistry, yet ozone is not new at all. Ozone therapy is already a major treatment modality in Europe, South America and a number of other countries. What is provided here will not be an exhaustive scientific treatise so much as a brief general introduction into what dentists are now doing with ozone therapies and the numerous oral/systemic links that make this subject so important for physicians so that, ultimately, they may serve their patients more effectively and productively.



http://www.ncbi.nlm.nih.gov/pubmed/25221698
Ann Med Health Sci Res. 2014 Jul;4(4):526-31.
doi: 10.4103/2141-9248.139301.


Qualitative analyses of the antimicrobial effect of ozonated water on supragingival plaque and salivary microbes.

Sadatullah S, Mohamed N, Razak F.

Abstract

BACKGROUND:

Ozone is an unstable gas, capable of oxidizing any biological entity. It is an effective bactericide in its gaseous as well as aqueous form.

AIMS:

The objective of this study was to determine the in-situ antimicrobial effect of 0.1 ppm ozonated water on plaque and salivary microorganisms.

SUBJECTS AND METHODS:

24 h old supragingival plaque (SP) was collected from the two most posterior teeth in the contralateral quadrants before and after a 30 s rinse with either distilled water (control group) or 0.1 ppm ozonated water (test group). Simultaneously stimulated whole saliva (SWS) was collected for both groups. The SP and SWS were stained with LIVE/DEAD Baclight Bacteria Viability fluorescent kit to visualize live and dead microbes. The salivary flow rate, calcium and protein level were also determined in the pre- and post-rinsed samples. The data obtained was subjected to One Way ANOVA test using Minitab 14 statistical software (PA, USA).

RESULTS:

0.1 ppm ozonated water was found effective in reducing the bacterial load in both the 24 h plaque and SWS samples, but it did not eliminate them completely. In addition, there was no statistically significant effect of the ozonated water rinse on the salivary flow rate, salivary calcium and protein concentration.

CONCLUSIONS:

0.1 ppm ozonated water rinse can be an effective adjunct to tooth brushing and flossing to maintain plaque and salivary bacterial load.



http://www.ncbi.nlm.nih.gov/pubmed/22574088
J Med Life. 2012 Feb 22;5(1):59-67. Epub 2012 Mar 5.

Ozone therapy in periodontics.

Gupta G, Mansi B.

Abstract

Gingival and Periodontal diseases represent a major concern both in dentistry and medicine. The majority of the contributing factors and causes in the etiology of these diseases are reduced or treated with ozone in all its application forms (gas, water, oil). The beneficial biological effects of ozone, its anti-microbial activity, oxidation of bio-molecules precursors and microbial toxins implicated in periodontal diseases and its healing and tissue regeneration properties, make the use of ozone well indicated in all stages of gingival and periodontal diseases. The primary objective of this article is to provide a general review about the clinical applications of ozone in periodontics. The secondary objective is to summarize the available in vitro and in vivo studies in Periodontics in which ozone has been used. This objective would be of importance to future researchers in terms of what has been tried and what the potentials are for the clinical application of ozone in Periodontics.



http://www.ncbi.nlm.nih.gov/pubmed/24324309
J Int Oral Health. 2013 Oct;5(5):79-84. Epub 2013 Oct 26.

Effect of Ozonised water on Chronic Periodontitis - A Clinical Study.

Katti SS1, Chava VK.

Abstract

BACKGROUND:

The aim of the study was to study the clinical effects of ozonated water on periodontal tissues.

MATERIALS & METHODS:

In the present study 30 subjects were selected with age ranging from 20 to 60 yearsand pocket depth of =5mm. Two sites were selected in each patient whichwere divided into two groups. Group 1(control group-irrigation with saline) and Group 2(study group-irrigation with ozonized water) and clinical parameters were recorded at baseline, 15 days and 30 days.

RESULTS:

When the comparison of mean values of Plaque Index and Gingival Index between the groups and at different time intervals were made, statistically significant difference were observed at 30 days at 5% level. When the mean values of clinical attachment level on mesial and distal site was compared between the groups, statistical significance was observed at 5% level and 1% level respectively.Similarly statistical significance at 5% level was observed at 15 and 30 days on buccal site.

CONCLUSION:

Subgingival irrigation with ozonized water is beneficial adjunct treatment modality to enhance periodontal health with significant role in periodontal therapy. How to cite this article: Katti SS, Chava VK. Effect of Ozonised water on Chronic Periodontitis - A Clinical Study. J Int Oral Health 2013;5(5):79-84.



http://www.ncbi.nlm.nih.gov/pubmed/23946585
J Pharm Bioallied Sci. 2013 Jun;5(Suppl 1):S89-94. doi: 10.4103/0975-7406.113304.

Application of ozone in the treatment of periodontal disease.

Srikanth A, Sathish M, Sri Harsha AV.

Abstract

Gingivitis and periodontitis are most common inflammatory diseases of supporting tissues of teeth. Role of microbial etiology and host response in progression of gingival and periodontal diseases has been well established. Because of the beneficial biological effects of ozone, due to its antimicrobial and immunostimulating effect, it is well indicated in the treatment of gingival and periodontal diseases. The objective of this article is to provide a general review about clinical applications of ozone in treatment of periodontal diseases and to summarize the available in vitro and in vivo studies in Periodontics in which ozone has been used.



http://www.ncbi.nlm.nih.gov/pubmed/16393498

Prim Dent Care. 2006 Jan;13(1):37-41.

The use of ozone in dentistry and medicine. Part 2. Ozone and root caries.

Baysan A, Lynch E.

Abstract

A previous paper, recently published in Primary Dental Care, gave an overview of the medical uses of ozone and outlined some of its uses in dentistry. The current paper focuses on a description of use of ozone in the management of root caries and considers recent studies in this area. There has been relatively limited research into the non-invasive (pharmaceutical) management of root caries. The best management strategy still remains to be developed. Initial studies have indicated that an application of ozone for a period of either 10 or 20 seconds is capable of clinically reversing leathery root carious lesions. It is suggested that, subject to confirmation from extensive trials, this simple and non-invasive technique may benefit many patients with root caries throughout the world since this approach to treat root caries can easily be employed in primary care clinics and in the domiciliary treatment of home-bound elderly people and immobile patients in hospices and hospitals.



http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3662033/
Iran Endod J. 2013 Spring; 8(2): 40–43.
http://www.ncbi.nlm.nih.gov/pubmed/23717326

A Review of the Properties and Applications of Ozone in Endodontics: An Update

Zahed Mohammadi, Sousan Shalavi, Mohammad Karim Soltani, and Saeed Asgary

Abstract

Ozone is a triatomic molecule consisting of three oxygen atoms. It is applied to oral tissues in the forms of ozonated water, ozonated olive oil and oxygen/ozone gas. This paper presents a brief review on the chemistry of ozone as well as its medical and dental applications focusing on its use in endodontics. Ozone’s antimicrobial activity, its effect on dentin bonding, toxicity and contra-indications are also reviewed.
Keywords: Antimicrobial, Dentin Bonding, Endodontics, Ozone, Toxicity

1.Introduction

In 1839, Christian F. Schonbein, first noticed the emergence of a pungent gas with an electric smell. Later, in 1857, Wener Von Siemens designed an ozone generator. Oxygen/ozone therapy has a long history of research and clinical and therapeutic applications on humans. The first medical application was in 1870 when Lender purified blood in test tubes. Medical applications became widespread throughout Europe and America. As of 1929, more than 114 diseases were listed for treatment with oxygen/ozone. In 1930, Fisch, used ozone on a regular basis in his dental practice in Switzerland (1).

2.Chemistry of ozone

Ozone (O3) is a triatomic molecule with three oxygen atoms and molecular weight of 47.98 g/mol. Thermodynamically, this molecule is a highly instable compound that decomposes to pure oxygen with a short half-life in particular temperature and pressure conditions (2). Ozone is 1.6-fold denser and 10-fold more soluble in water (49 mL in 100 mL water at 0°C) than oxygen. Although ozone is not a radical molecule, it is the third most potent oxidant after fluorine and per sulfate. Ozone is an unstable gas that cannot be stored and should be used at once because it has a half-life of 40 min at 20°C (2). Ozone is naturally produced by the photo dissociation of molecular oxygen (O2) into activated oxygen atoms, which then react with further oxygen molecules. This transient radical anion rapidly becomes protonated, generating hydrogen trioxide (HO3), which, in turn, decomposes to an even more powerful oxidant, the hydroxyl radical (OH) (3). It is the fundamental form of oxygen that occurs naturally as a result of ultraviolet energy or lightning, causing a temporary recombination of oxygen atoms into groups of three. In the clinical setting, an oxygen/ozone generator simulates lightning via an electrical discharge field. Ozone gas has a high oxidation potential and is 1.5 times more effective than chloride when used as an antimicrobial agent against bacteria, viruses, fungi, and protozoa. It also has the capacity to stimulate blood circulation and the immune response. Ozone has been indicated for the treatment of 260 different pathologies (4, 5).

3.Routes of ozone therapy

Ozone is applied to oral tissues in the following forms: Ozonated water, ozonated olive oil, and oxygen/ozone gas. Ozonated water and olive oil have the capacity to entrap and then release oxygen/ozone; an ideal delivery system. These forms of application are used individually or in combination to treat dental disease (6).

4.Applications of ozone in medicine

Most of our knowledge is based on multiple case reports from hospitals and clinics. Ozone can enhance both lung function and inflammatory airway responses to inhale allergen in cases with pre-existing allergic airway diseases (7).

Medicated forms of O3 in a gaseous form are somewhat unusual, and that is why special application techniques have had to be developed for the safe use of ozone. In other words, due to the difficulty in handling and administering gaseous ozone, some methods and devices have been introduced to enhance its effectiveness. In local applications such as the treatment of external wounds, transcutaneous O3 gas bath has been established as a most practical and useful method; at low (subatmospheric) pressure in a closed system guaranteeing no escape of ozone into the ambient air. Ozonized water, whose use is particularly known in dental medicine, is optimally applied as a spray or in compressed form (8).

Apart from rectal insufflation which is principally used for the treatment of intestinal conditions, and also applied systemically, autohemotherapy has established itself as a systemic therapy of choice. A corresponding dosage of ozone gas is transferred (in the form of microbubbles) into 50 to 100 ml of the patient’s blood in a sealed, pressure-less system, thus achieving the finest possible distribution to reach the greatest possible number of red and white blood cells to activate their metabolism. This is a markedly low-risk method when hygiene guidelines are observed, disposable units are used, and the material used is ozone-resistant. In the therapy of pain in the locomotor system, ozone can be applied supportively in the form of intramuscular or intra-articular injections (2).

5.Ozone in dentistry

Muller et al. (9) compared the influence of ozone gas with photodynamic therapy (PDT) and known antiseptic agents (2% Chlorhexidine, 0.5 and 5% sodium hypochlorite solutions) on a multispecies oral biofilm in-vitro. The studied bacteria were Actinomyces naeslundii, Veillonella dispar, Fusobacterium nucleatum, Streptococcus (S.) sobrinus, S. oralis and Candida (C.) albicans. Gasiform ozone was produced by vacuum ozone delivery system Kavo HealOzone (Biberach, Germany). They concluded that the matrix-embedded microbial populations in biofilm are well protected against antimicrobial agents. Only 5% NaOCl solution was able to eliminate all bacteria effectively. Gasiform ozone or PDT was not able to significantly reduce or completely eliminate bacteria in the biofilm (9). Baysan et al. assessed antimicrobial effect of Kavo HealOzone device on primary root caries lesions (PRCL) and evaluated the efficiency of ozone specifically on S. mutans and S. sobrinus (10). As a result, ozone exposure to either 10 or 20 s under experimental conditions reduced the total levels of microorganisms in the PRCLs to <1% of the control values. Application of ozone for a period of 10 s was also capable of reducing the numbers of S. mutans and S. sobrinus in-vitro (10). Holmes (11) observed the effect of KaVo Healozone device on PRCL followed by professionally-applied remineralizing solution containing xylitol, fluoride, calcium, phosphate and zinc. This treatment modality was applied to 89 patients over 60. After 18 months 100% of ozone-treated primary root caries lesions (PRCLs) had improved. In control group, where lesions were left without treatment, only one PRCL had improved. In 62% of cases the status remained leathery, while in 37% of PCRL’s had worsened from leathery to soft texture (11).

The influence of ozonated water on the epithelial wound healing process in the oral cavity was observed by Filippi (12). It was found that ozonized water applied on a daily basis can accelerate the healing rate in oral mucosa. This effect was seen in the first two postoperative days. Daily treatment with ozonized water accelerates the physiological healing rate in the treated wounds as such changes were not seen in the untreated wounds (12).

6.Ozone in endodontics

Ozone gas in a ~4 g/m3 concentration (HealOzone; KaVo, Biberach, Germany) is used clinically for endodontic treatments.

6.1.Antimicrobial activity

Most studies on the applications of ozone in endodontics focus on its antimicrobial activity. Nagayoshi et al. (13) found that ozonated water (0.5–4 mg/L) was highly effective in killing both gram positive and negative micro-organisms. Gram negative bacteria, such as Porphyromonas (P.) endodontalis and P. gingivalis were substantially more sensitive to ozonated water than gram positive oral streptococci and C. albicans in pure culture. Hems et al. (14) evaluated the potential of ozone as an antibacterial agent using Enterococcus (E.) faecalis as a test species. Ozone was used both as gasiform (produced by Pure zone device), and aqueous (optimal concentration 0.68 mg/L). They concluded that ozone in solution was antibacterial against planktonic E. faecalis after 240 s treatment. However it was not effective against E. faecalis within a biofilm unless they were displaced into the surrounding medium by agitation (14).

Estrela et al. (15) studied antimicrobial effects of ozonated water, gaseous ozone and antiseptic agents (2.5% hypochlorite and 2% chlorhexidine) in infected human dental root canals. Over the 20 min contact time none of these substances had antibacterial effect against E. faecalis in the infected root canals.

Thanomsub et al. (16) tested the effects of ozone treatment on cell growth and ultrastructural changes in bacteria (Escherichia coli, Salmonella sp., Staphylococcus aureus and Bacillus subtilis). It was discovered that ozone at 0.167 mg/min/L concentration can be used to sterilize water, which is contaminated within 30 min with up to 105 cfu/ml bacteria. Destroying of bacterial cell membrane was observed, subsequently producing intercellular leakage and eventually causing cell lysis. Nevertheless, these ozone concentrations have no significant effect on the cell viability of bacterial cultures at higher concentrations of 106 and 107 CFU/mL (16).

Polydorou et al. (17) found that 80 s as well as 40s applications of ozone was effective in eliminating S. mutans. In an animal study on the infected dog's teeth, Silveira et al. (2007) showed 77% root canal treatment success rate when using ozonized oil as an intra-canal medicament. A further study evaluated the effectiveness of ozonated water in the elimination of C. albicans, E. faecalis, and endotoxins from root canals (18). Findings revealed that ozonated water was effective against both C. albicans and E. faecalis immediately after treatment; however it did not have substantivity. Furthermore, ozonated water demonstrated no anti-endotoxin activity. The disinfecting effect of ozonized oxygen (120 s from the HealOzone) on E. faecalis has been assessed (19). Findings revealed that ozonized oxygen appears to be suitable for disinfecting root canal systems in cases where sodium hypochlorite is not indicated. Huth et al. (20) assessed the effectiveness of aqueous and gaseous ozone against E. faecalis, C. albicans, Peptostreptococcus micros and Pseudomonas aeruginosa cultured in planktonic media or in mono-species biofilms. Results demonstrated that high-concentrated gaseous and aqueous ozone was dose-, strain- and time-dependently effective against the tested microorganisms. The antibacterial activity of gaseous ozone was shown to be greater than KTP laser and less than NaOCl (21) and ozone gas delivered into irrigating fluids in the root canal may be useful as an adjunct for endodontic disinfection (22).

7.Effects on dentin bonding

Schmidlin et al. (23) evaluated the influence of direct high-dose gaseous ozone application (2100 ppm) on dentin and enamel shear bond strength; despite a possible retention of surface and subsurface oxide-related substances during high-dose ozone application, the strength was not impaired. Thus, adhesive restoration placement should be possible immediately after ozone application for cavity disinfection. However, other researchers reported that adhesion of the self-adhesive resin cement RelyX Unicem (3M ESPE, Seefeld, Germany) was significantly reduced after using gaseous ozone (24).

Magni et al. (25) indicated that Ozone gas did not compromise the mechanical properties of the adhesives [including Prime & Bond NT (Dentsply), Excite (Ivoclar-Vivadent), Syntac/Heliobond (Ivoclar-Vivadent) and Silorane System Adhesive (3 M-ESPE)]. When ozone gas was used to disinfect the cavity before a restoration, it had no influence on immediate enamel and dentin bond strength (26). Çehreli et al. (27) revealed that pre-treatment with ozone improved the marginal sealing ability of the fissure sealants; another study demonstrated that ozone therapy improved shear bond strength of AH-26 and EX Fill root canal sealers (28). Gurgan et al. (29) showed that ozone treatment did not impaired the shear bond strength of two self-etch adhesives (Clearfil SE Bond and Clearfil Tri-S Bond) used on coronal and radicular dentin. However other studies showed that ozone decreased the microtensile bond strength of dentin-composite resin interface (30) and reduced the initial microtensile bond strength of Clearfil SE Bond (31). According to Arslan et al. (32) ozone did not significantly affect the dentin bond strength of a silorane-based resin composite, filtek supreme. Also, another study revealed that ozone gas and ozonated water had no deleterious effects on the bond strengths and interfaces (33).

8.Toxicity

Ozone inhalation can be toxic to the pulmonary system and other organs. Complications caused by ozone therapy are infrequent at 0.0007 per application. Known side-effects are epiphora, upper respiratory irritation, rhinitis, cough, headache, occasional nausea, vomiting, shortness of breath, blood vessel swelling, poor circulation, heart problems and even stroke. Because of ozone's high oxidative power, all materials that come in contact with the gas must be ozone resistant, such as glass, silicon, and Teflon. However, in the event of ozone intoxication, the patient must be placed in the supine position and treated with vitamin E and N-acetylcysteine (2).

9.Contra-indications of ozone therapy

The following conditions mentioned in the medical literatures contraindicate ozone use e.g. acute alcohol intoxication, recent myocardial infarction, hemorrhage in any organ, pregnancy, hyperthyroidism, thrombocytopenia and ozone allergy (2-4).

10.Conclusion

Ozone is applied to oral tissues in various forms: ozonated water, ozonated olive oil, and oxygen/ozone gas. Ozone has been used in medicine extensively. In dentistry, its effectiveness on wound healing, antibacterial activity, and its effect on dentin bonding has been investigated. Ozone improves wound healing, assists in treating root caries and can be used against endodontic microbiota. Furthermore, it seems that ozone does not have significant adverse effect on dentin bonding. In spite of infrequency of side effects, ozone therapy may cause serious medical complications if incorrectly used. Therefore care must be taken when handling ozone.

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14. Hems RS, Gulabivala K, Ng YL, Ready D, Spratt DA. An in vitro evaluation of the ability of ozone to kill a strain of Enterococcus faecalis. Int Endod J. 2005;38(1):22–9. doi: 10.1111/j.1365-2591.2004.00891.x. [PubMed] [Cross Ref]

15. Estrela C, Estrela CR, Decurcio DA, Hollanda AC, Silva JA. Antimicrobial efficacy of ozonated water, gaseous ozone, sodium hypochlorite and chlorhexidine in infected human root canals. Int Endod J. 2007;40(2):85–93. doi: 10.1111/j.1365-2591.2006.01185.x. [PubMed] [Cross Ref]

16. Thanomsub B, Anupunpisit V, Chanphetch S, Watcharachaipong T, Poonkhum R, Srisukonth C. Effects of ozone treatment on cell growth and ultrastructural changes in bacteria. J Gen Appl Microbiol. 2002;48(4):193–9. [PubMed]

17. Polydorou O, Pelz K, Hahn P. Antibacterial effect of an ozone device and its comparison with two dentin-bonding systems. Eur J Oral Sci. 2006;114(4):349–53. doi: 10.1111/j.1600-0722.2006.00363.x. [PubMed] [Cross Ref]

18. Cardoso MG, de Oliveira LD, Koga-Ito CY, Jorge AO. Effectiveness of ozonated water on Candida albicans, Enterococcus faecalis, and endotoxins in root canals. Oral Surg Oral Med Oral Pathol Oral Radiol Endod. 2008;105(3):e85–91. doi: 10.1016/j.tripleo.2007.10.006. [PubMed] [Cross Ref]

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20. Huth KC, Jakob FM, Saugel B, Cappello C, Paschos E, Hollweck R, Hickel R, Brand K. Effect of ozone on oral cells compared with established antimicrobials. Eur J Oral Sci. 2006;114(5):435–40. doi: 10.1111/j.1600-0722.2006.00390.x. [PubMed] [Cross Ref]

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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3307081/
J Med Life. 2012 Feb 22; 5(1): 59–67. 2012 Mar 5.
PMCID: PMC3307081

Ozone therapy in periodontics

G Gupta and B Mansi

Abstract

Gingival and Periodontal diseases represent a major concern both in dentistry and medicine. The majority of the contributing factors and causes in the etiology of these diseases are reduced or treated with ozone in all its application forms (gas, water, oil). The beneficial biological effects of ozone, its anti-microbial activity, oxidation of bio-molecules precursors and microbial toxins implicated in periodontal diseases and its healing and tissue regeneration properties, make the use of ozone well indicated in all stages of gingival and periodontal diseases. The primary objective of this article is to provide a general review about the clinical applications of ozone in periodontics. The secondary objective is to summarize the available in vitro and in vivo studies in Periodontics in which ozone has been used. This objective would be of importance to future researchers in terms of what has been tried and what the potentials are for the clinical application of ozone in Periodontics.

Introduction

The word ozone comes from the Greek “ozein” meaning odorant. Ozone (also known as triatomic oxygen and trioxygen) is an allotropic form of oxygen occurring naturally in the Earth’s atmosphere. It surrounds the Earth at an altitude of between 50,000 and 100,000 feet. [1] It is created in nature when ultraviolet rays cause oxygen atoms to temporarily recombine in groups of three. It is also formed by the action of electrical discharges on oxygen, so it is often created by thunder and lightning.

It has got the capacity to absorb the harmful ultra-violet rays present in the light spectrum from the Sun. It is a pale blue gas that condenses to a deep blue liquid at very low temperatures.

Ozone is an unstable gas and it quickly gives up nascent Oxygen molecule to form Oxygen gas. Due to the property of releasing nascent Oxygen, it has been used in human medicine since long back to kill bacteria, fungi, to inactivate viruses and to control hemorrhages. [2] Medical grade ozone is made from pure medical oxygen. It is produced commercially in ozone generators, which involves sending an electrical discharge through a specially-built condenser containing oxygen.

History

First discovered until 1840 by the German chemist Christian Frederick Schonbein at the University of Basil in Switzerland, ozone was first used in medicine in 1870 by Landler. However, it was not until 1932 that ozone was seriously studied by the scientific community, when ozonated water was used as a disinfectant by Dr. E. A. Fisch, [3] a Swiss dentist. Fisch had the first idea to use ozone as either a gas or ozonated water in his practice. By a twist of fate, a surgeon, Dr. E Payr (1871–1946) had to be treated for a gangrenous pulpite and remained astonished by the result achieved with local ozone treatment. He enthusiastically extended its application to general surgery.

At the time, ozone therapy was difficult and limited due to the lack of ozone-resistant materials, such as Nylon, Dacron, and Teflon, until 1950 when ozone-resistant materials were manufactured. At that time Joachim Hänsler, a German physicist and physician, joined another German physician, Hans Wolff, to develop the first ozone generator for medical use. Their design continues to be the basis for modern equipment.

Ozone generators

There are three different systems for generating ozone gas: [4]

• Ultraviolet System: produces low concentrations of ozone, used in esthetics, saunas, and for air purification.

• Cold Plasma System: used in air and water purification.

• Corona Discharge System: produces high concentrations of ozone. It is the most common system used in the medical/ dental field. It is easy to handle and it has a controlled ozone production rate.

Medical grade ozone is a mixture of pure oxygen and pure ozone in the ratio of 0.05% to 5% of 3 and 95% to 99.95% of O2. Due to the instability of the 3 molecule, medical grade ozone must be prepared immediately before use. Within less than an hour after preparation only half of the mixture is still ozone while the other half is transformed into oxygen. As a result, it is impossible to store ozone over long periods of time. In order to control the decomposition of 3 into oxygen it can be associated with a vehicle with aqueous properties to promote the conversion more quickly or with a vehicle with more viscous properties to retard the conversion.

Use of ozone in dentistry

The use of ozone in dentistry is gaining its place in every day's dental practice and is used in almost all dental applications. The undisputed disinfection power of ozone over other antiseptics makes the use of ozone in dentistry a very good alternative and/or an additional disinfectant to standard antiseptics.

Due to safety concerns, 3 gas was not recommended for intra-oral use. Only dissolved ozone in water and ozonated oils were and are still commonly used in different fields of dentistry. With the development of a foot pedal-activated dental handpiece with a suction feature, 3 gas can now be used safely in situations where diffusion is an important factor, i.e. dental hard tissues.

According to German dentist Fritz Kramer, [5] ozone, such as in the form of ozonated water, can be used in the following ways.

1. as a powerful disinfectant

2. in its ability to control bleeding

3. in its ability to cleanse wounds in bones and soft tissues.

4. by increasing the local supply of oxygen to the wound area, ozone can improve healing.

5. ozonated water can increase temperature in the area of the wound, and this increase the metabolic processes related to wound healing.

Dr. Kramer points out that ozonated water can be used in a number of different ways:

1. as a mouth rinse (especially in cases of gingivitis, paradentosis, thrush or stomatitis);

2. as a spray to cleanse the affected area, and to disinfect oral mucosa, cavities and in general dental surgery;

3. as an ozone/water jet to clean cavities of teeth being capped, receiving root canal therapy, and in treating painful gingivitis and stomatitis.

Biological actions

The application of ozone in dentistry comes as a result of physico-chemical properties: There are several known actions of ozone on human body, such as immunostimulating and analgesic, antihypoxic and detoxicating, antimicrobial, bioenergetic and biosynthetic (activation of the metabolism of carbohydrates, proteins, lipids) etc. [6]

1. Antimicrobial effect- Ozone works destructively against bacteria, fungi, and viruses. The antimicrobial effect of ozone is a result of its action on cells by damaging its cytoplasmic membrane due to ozonolysis of dual bonds and also ozone-induced modification of intracellular contents (oxidation of proteins loss of organelle function) because of secondary oxidants effects. This action is non-specific and selective to microbial cells; it does not damage human body cells because of their major antioxidative ability. Ozone is very efficient in antibiotics resistant strains. Its antimicrobial activity increases in liquid environment of the acidic pH. In viral infections the ozone action lies in the intolerance of infected cells to peroxides and change of activity of reverse transcriptase, which takes part in synthesis of viral proteins. [6] Being a very strong oxidant it joins with biomolecules containing cysterine, cysteine, methionine, histidine (all being part of bacterial cell membranes. The main targets of their attack are the thiol groups of the amino acid cysteine. As a result of the reaction of ozone with unsaturated fatty acids of a lipid sheath of a virus the lipid sheath of a virus melts. The research shows that a few-second-application of ozone stops all vital functions of bacteria which are incapable of developing any self-immunity to its action. Gram+ (Gram-positive) bacteria are more sensitive to the action of ozone than Gram– (Gram-negative) bacteria. Oxygen-free bacteria react to ozone as well. Among cariogenic bacteria Streptococcus mutans and Streptococcus sobrinus are the most sensitive. Ozone easily acts on multi unsaturated fatty acids which occur in virus sheaths. Ozone reacts also with ascorbinians and tocopherols. [7]

2. Immunostimulating Effect- Ozone influences cellular and humoral immune system. It stimulates proliferation of immunocompetent cells and synthesis of immunoglobulins. It also activates function of macrophages and increases sensitivity of micro-organisms to phagocytosis. [6] As a response to this activation through ozone, the body's immune cells produce special messengers called cytokines. These molecules in turn activate other immune cells, setting off a cascade of positive change throughout the immune system, which is stimulated to resist diseases. This means that the application of medical ozone is extremely useful for immune activation in patients with a low immune status and/or immune deficit. [8] Ozone causes the synthesis of biologically active substances such as interleukins, leukotrienes and prostaglandins which is beneficial in reducing inflammation and wound healing. [6] Ozone in high concentration causes immunodepressive effect whereas in its low concentration immunostimulating effect. [7]

3. Antihypoxic effect- Ozone brings about the rise of pO2 in tissues and improves transportation of oxygen in blood, which results in change of cellular metabolism – activation of aerobic processes (glycolysis, Krebs cycle, ß-oxidation of fatty acids) and use of energetic resources. Repeating low doses of ozone activate enzymes: super-oxide dismutases, catalases, dehydrogenase, and glutatione peroxidases. They are part of complex enzymatic systems which protect organisms against the action of oxygen-free radicals. It also prevents formation of erythrocytes aggregates and increases their contact surface for oxygen transportation. Its ability to stimulate the circulation is used in the treatment of circulatory disorders and makes it valuable in the revitalizing organic functions. [6] Ozone improves the metabolism of inflamed tissues by increasing their oxygenation and reducing local inflammatory processes. By changing the cell membrane structure of erythrocytes and causing the increase of its negative charge it influences the structure change as well as blood cell elasticity. This in consequence reduces blood cell rolling and enables blood flow in capillary vessels. By increasing the concentration of 2,3 Diphosphoglycerate (2,3-DPG), ozone changes the configuration of erythrocytes, which enables them to return oxygen in the inflamed tissue. [7]

4. Biosynthetic Effect- It activates mechanisms of protein synthesis, increases amount of ribosomes and mitochondria in cells. These changes on the cellular level explain elevation of functional activity and regeneration potential of tissues and organs. [6]

5. Ozone causes secretion of vasodilators such as NO, which is responsible for dilatation of arterioles and venules. [6] It also activates angiogenesis. [7]

6. Ozone, when acting on the organic substance of mineralized tooth tissues intensifies their remineralization potential. At the same time, it is capable of “opening” dentinal tubules, which enables the diffusion of calcium and phosphorus ions to the deeper layers of carious cavities. [9]

A high concentration of ozone kills bacteria very quickly and is thousand times more powerful than other bacterial killing agents. The average concentration of ozone used in treatments is 25 gm of ozone per milliliter of oxygen/ozone gas mixture that translates into 0.25 parts of ozone to 99.75 parts of oxygen. Evidence-based research has shown that at this concentration, ozone effectively kills bacteria, fungi, viruses and parasites. [10] As an antimicrobial agent, it is a powerful oxidizer at a dramatically lower concentration than chlorine with none of the toxic side effects. One molecule of ozone is equal to between 3,000 to 10,000 molecules of chlorine and it kills pathogenic organisms 3,500 times faster. [10] Studies have revealed that it only takes 10 sec to kill 99 % of bacteria, fungi and viruses. [11] It can oxidize many organic compounds and it is a powerful germicide. [12] Some of the other effects are circulatory enhancement, disruption of tumor metabolism and stimulation of oxygen metabolism. [13]

According to most authors, a 10-sec-application of ozone causes the destruction of 99% of bacteria, and a 20-sec-application even of 99.9%. In this way, so-called ecological niche appears.

However, it is not conducive to their repeated colonization within 4 to 6 weeks. [14,15] Ozone is not toxic when it is given in the amount of 0.05 ppm for 8 hours. During ozone therapy a maximum concentration of ozone in oral cavity amounts to 0.01 ppm.

Goals of ozone therapy

Setting the standard-of-care and therapeutic goals are based on sound evidence-based science is critical. Therapeutic goals are inclusive and not exclusive of standard of care. The goals of oxygen/ozone therapy are: [10]

1. Elimination of pathogens.

2. Restoration of proper oxygen metabolism.

3. Induction of a friendly ecologic environment.

4. Increased circulation.

5. Immune activation.

6. Simulation of the humoral anti-oxidant system.

Use in dentistry

Periodontology- Gingivitis, Periodontits, Periimplantitis, Surgical cuts, Prophylaxis

Dental and oral pathology- Caries, Enamel cracks, Root canal treatment, Tooth whitening, Dentinal hypersensitivity, Abscess, Granuloma, Fistulae, Apthae, Herpes infection, Stomatitis – Candidiasis

Surgery- Implantation, Re-plantation, Extraction, Wound Healing, Coaguloapathy - prolonged bleeding

Prosthodontics and restorative dentistry- Stumps and crown disinfection, Cavity disinfection

Orthodontics and orthopedics- TMJ dysfunctions, Trismus, Relaxation, Myoarthtopathy

Diagnostics- Vitality test

Appliances producing ozone for dental use

1. HealOzone by KaVo is air-based and the application of the gas takes place in a closed circuit. Its surplus is sucked out and neutralized by manganese ions. The concentration of ozone in the cap adjacent to the tissue amounts to 2100 ppm. Perfect air tightness of the cap is necessary for the application of ozone. Therefore, the application is only possible on the surfaces where such air tightness can be provided.

2. OzonyTron by MYMED Gmb H. - Oxygen activation generator (OzonytronX—Biozonix, München, Germany) uses the power of high frequency and voltage. Activated oxygen (ozone) concentration can be adjusted in 5 levels via current strength. Inside the glass probe, which is formed by a double glass camera, is a noble gasses mixture that is conducting and emitting electromagnetic energy. When the tip of the probe gets in contact with the body it emits energy around the treated area and splits environmental diatomic oxygen in singular atomic oxygen and ozone. The concentration of ozone in the operation field is 10 to 100 µg/ml (becomes a fungi-, viru-, and bacteriocide at the intensity of 1–5 µg/ml). There is no closed circuit here, therefore, ozone can be applied to the places that are difficult to reach, e.g. gingival pockets or root canals.

3. Product photo (Prozone) by W&H - It is characterized by its ease of use and safety of application (preset tissue-compatible dosages in the indication areas of periodontitis and endodontitis). Prozone ensures a hygienic procedure during the gassing of the pockets due to its exchangeable plastic attachments (Perio tips or Endo tips).

Route of ozone administration

1. Gaseous Ozone - Gaseous ozone is most frequently used in restorative dentistry and endodontics. Topical administration of the gaseous form can be via an open system or via a sealing suction system as a prerequisite to avoid inhalation and adverse effects. Ozone appears to be an integral part of noninvasive therapy of dental caries, as a disinfectant prior to placing a direct restoration and as therapy for hypomineralized teeth. [16]

2. Ozonated Water - Ozonated water has been shown to be very effective against bacteria, fungi and viruses and is also less expensive compared to other chemical cleaners. [17] Gaseous ozone was shown to be a more effective microbicide than the aqueous form and, applied for 3 min, may be used as a dental disinfectant. [18] Because ozone gas has been found to have toxic effects if inhaled into the respiratory tract, [16,18,19] ozonated water may be useful to control oral infections and various pathogens.[15]

3. Ozonized Oil - In addition to ozone application in its gaseous and aqueous form, sunflower ozonized oil also seems extremely convenient. The wide accessibility of sunflower oil makes it a competitive antimicrobial agent. Ozonized oil (Oleozone, Bioperoxoil) has shown to be effective against Staphylococci, Streptococci, Enterococci, Pseudomonas, Escherichia coli and especially Mycobacteria [16,20,21] and has been utilized for the cure of fungal infections. [16,20]

Ozone therapy in periodontics

The main use of ozone in dentistry relies on its antimicrobial properties. It is proved to be effective against both Gram positive and Gram negative bacteria, viruses and fungi. [22]

Ebensberger et al [23] evaluated the effect of irrigation with ozonated water on the proliferation of cells in the periodontal ligament adhering to the root surfaces of 23 freshly extracted completely erupted third molars. The teeth were randomly treated by intensive irrigation with ozonated water for 2 min or irrigation with a sterile isotonic saline solution, serving as a control group. The periodontal cells of these teeth were studied immunohistochemically to mark proliferating cell nuclear antigen (PCNA). It was observed that the labeling index (the number of positive cells compared to the total number of cells suggesting enhancement of metabolism) was higher among the teeth irrigated with ozone (7.8% vs. 6.6%); however, the difference was not statistically significant (p = 0.24). They concluded that the 2 min irrigation of the avulsed teeth with non-isotonic ozonated water might lead not only to a mechanical cleansing, but also decontaminate the root surface, with no negative effect on periodontal cells remaining on the tooth surface.

Nagayoshi et al [24] examined the effect of ozonated water on oral microorganisms and dental plaque. Dental plaque samples were treated with 4mL of ozonated water for 10 s. they observed that ozonated water was effective for killing gram-positive and gram-negative oral microorganisms and oral Candida albicans in pure culture as well as bacteria in plaque biofilm and therefore might be useful to control oral infectious microorganisms in dental plaque.

Nagayoshi et al [17] tested the efficacy of three different concentrations of ozone water (0.5, 2, and 4 mg/ml in distilled water) on the time-dependent inactivation of cariogenic, periodontopathogenic and endodontopathogenic microbes (Streptococcus, Porphyromonas gingivalis and endodontalis, Actinomyces actinomycetemcomitans, Candida albicans) in culture and in biofilms. They confirm that ozonated water was highly effective in killing of both gram positive and gram negative micro-organisms.

Depending on the dosage, the oral microbes were inactivated after 10 seconds. Gram negative anaerobes, such as Porphyromonas endodontalis and Porphyromonas gingivalis were substantially more sensitive to ozonated water than gram positive oral streptococci and Candida albicans in pure culture. Furthermore ozonated water had strong bactericidal activity against bacteria in plaque biofilm. In addition, ozonated water inhibited the accumulation of experimental dental plaque in vitro.

Ramzy et al [24] irrigated the periodontal pockets by ozonized water in 22 patients suffering from aggressive periodontitis (age range from 13 to 25 years). Periodontal pockets were irrigated with 150 ml of ozonized water over 5 to 10 minutes once weekly, for a clinical four weeks study, using a blunt tipped sterile plastic syringe. High significant improvement regarding pocket depth, plaque index, gingival index and bacterial count was recorded related to quadrants treated by scaling and root planning together with ozone application. They also reported significant reduction in bacterial count in sites treated with ozonized water.

Huth et al [19] in their study declared that the aqueous form of ozone, as a potential antiseptic agent, showed less cytotoxicity than gaseous ozone or established antimicrobials (chlorhexidine digluconate-CHX 2%, 0.2%; sodium hypochlorite-NaOCl 5.25%, 2.25%; hydrogen peroxide-H2O2 3%) under most conditions. Therefore, aqueous ozone fulfils optimal cell biological characteristics in terms of biocompatibility for oral application.

Huth et al [26] in their later paper examined the effect of ozone on the influence on the host immune response. These researchers chose the NF-kappaB system, a paradigm for inflammation-associated signalling/transcription. Their results showed that that NF-kappaB activity in oral cells in periodontal ligament tissue from root surfaces of periodontally damaged teeth was inhibited following incubation with ozonized medium. The Huth 2007 study establishes a condition under which aqueous ozone exerts inhibitory effects on the NF-kappaB system, suggesting that it has an anti-inflammatory capacity.

Muller et al [27] compared the influence of ozone gas with photodynamic therapy (PDT) and known antiseptic agents (2% Chlorhexidine, 0,5 and 5% hypochlorate solutions) on a multispecies oral biofilm in vitro. The following bacteria were studied – Actinomyces naeslundii, Veillonella dispar, Fusobacterium nucleatum, Streptococcus sobrinus, Streptococcus oralis and Candida albicans. Gasiform ozone was produced by vacuum ozone delivery system Kavo Healozone. They concluded that the matrix-embedded microbial populations in biofilm are well protected towards antimicrobial agents. Only 5 % Hypochlorate solution was able to eliminate all bacteria effectively. Usage of gasiform ozone or PDT was not able to reduce significantly or completely eliminate bacteria in the biofilm.

Kronusová [28] used ozone in following cases: prevention of dental caries in fissures of the first permanent molars in children, application of ozone in prepared cavity, after tooth extraction, in case of postextractional complications, in patients with chronic gingivitis, periodontitis and periodontal abscesses, herpes labialis, purulent periodontitis, dentition difficilis, etc. Almost all patients with gingivitis showed subjective and objective improvement of their status, as well as patients with periodontal abscess, where no exsudation was observed. Application of ozone after tooth extraction was found also quite useful – only 10 % of patients suffered from such complication as alveolitis sicca, but even in these cases the clinical course was shorter and more moderate.

The influence of ozonized water on the epithelial wound healing process in the oral cavity was observed by Filippi. [29] It was found that ozonized water applied on the daily basis can accelerate the healing rate in oral mucosa. This effect can be seen in the first two postoperative days. The comparison with wounds without treatment shows that daily treatment with ozonized water accelerates the physiological healing rate.

In the study by Karapetian et al, [30] periimplantitis treatment with conventional, surgical and ozone therapy methods was investigated, and it was found that the most effective bacteria reduction was in the ozone-treated patient group. The authors concluded that the main challenge seems to be the decontamination of the implant surface, its surrounding tissue and the prevention of recolonization with periodontal pathogenic bacteria.

Kshitish and Laxman [31] conducted a randomized, double-blind, crossover split-mouth study on 16 patients suffering from generalized chronic periodontitis. The study period of 18 days was divided into two time-intervals, i.e. baseline (0 days) to 7th day, with a washout period of 4 days followed by a second time interval of 7 days.

Subgingival irrigation of each half of the mouth with either ozone or chlorhexidine was done at different time intervals. They observed a higher percentage of reduction in plaque index (12%), gingival index (29%) and bleeding index (26%) 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 3 and chlorhexidine, there was no antibacterial effect on Porphyromonas gingivalis (Pg) and Tannerella forsythensis. The antifungal effect of ozone from baseline (37%) to 7th day (12.5%) was pronounced during the study period, unlike CHX, which did not demonstrate any antifungal effect. No antiviral property of ozone was observed. The antiviral efficacy of chlorhexidine was better than that of ozone. They concluded that despite the substantivity of chlorhexidine, the single irrigation of ozone is quite effective to inactivate microorganisms.

Application modalities

According to the clinical case, different applications modalities are available using ozone gas, irrigation with ozonated water and in-office use of ozonized oil as well as home use.

Gas application via a customized thermoformed dental appliance- A customized suckdown thermoformed hard or medium-soft dental appliance can be prepared. It should extend 2-3 mm beyond the affected gingival area, leaving a free space for gas circulation. 2 ports should be attached for the gas inlet and outlet respectively at the distal and mesial of the treatment area. The edges of the appliance should be reclined with light or medium body silicone. Light-cured dam can also be applied as an extra safety precaution to completely seal the borders. The ports to the generator and the suction pump should then be attached. This procedure will treat both hard and soft tissues of the affected area. PVC or silicone cap can be used to treat individually all the indicated areas in difficult situations where such an appliance is hard to use or uncomfortable to the patient.

Irrigation with Ozonated Water- Ozonated water can be used to irrigate the affected area during and after scaling, root surface planning, and non-surgical pocket curettage.

In-office and Home Use of Ozonized Olive Oil- After in-office treatment with ozone gas or ozonated water, pockets can be filled with ozonized olive oil using a blunt 25G needle or any other appropriate tip. Patient can be given some of the oils for home use. In-office ozonized oil application can be repeated once a week.

Surgical Procedures- Ozonated water can be used as an irrigant during the surgical procedure and/or as a final surgical site lavage. The sutures can be covered with a thin layer of ozonized oil and the patient can be instructed to apply the oil 3-4 times a day.

Peri-Implantitis- Peri-implantitis is very bothering to both the dentist and the patient. After thorough assessment and if a decision is taken to salvage the case, different modes of therapy are used in order to save the implant from total loss. Laser and/or manual debridement along with antiseptic solutions and topical anti-microbial medicines are commonly performed with a varying degree of success. Ozone can play an important role and be used as gas or in aqueous form. An appropriate length of PVC or silicone cap can be cut to cover the abutment fully. It should properly seal the gingival borders around the implant. Ozone gas infiltrations can also be used in this situation. Ozonated water can be used as an irrigant during debridement and curettage. Patient can be advised to apply ozonized oil on the treated area 3-4 times/day.
Desensitization of sensitive root necks [2]

Quick and prompt relief from root sensitivity has been documented after ozone spray for 60 seconds followed by mineral wash onto the exposed dentine in a repetitive manner. This desensitization of dentine lasts for longer period of time. Smear layer present over the exposed root surface prevents the penetration of ionic Calcium and Fluorine deep into the dentinal tubules. Ozone removes this smear layer, opens up the dentinal tubules, broadens their diameter and then Calcium and Fluoride ions flow into the tubules easily, deeply and effectively to plug the dentinal tubules, preventing the fluid exchange through these tubules. Thus, ozone can effectively terminate the root sensitivity problem within seconds and also lasts longer than those by conventional methods.

Ozone toxicity

Ozone inhalation can be toxic to the pulmonary system and other organs. Complications caused by ozone therapy are infrequent at 0.0007 per application. Known side-effects are epiphora, upper respiratory irritation, rhinitis, cough, headache, occasional nausea, vomiting, shortness of breath, blood vessel swelling, poor circulation, heart problems and at times stroke. [44] Because of ozone's high oxidative power, all materials that come in contact with the gas must be ozone resistant, such as glass, silicon, and Teflon. However, in the event of ozone intoxication the patient must be placed in the supine position, and treated with vitamin E and n-acetylcysteine. [16]

Contraindications


The following are contraindications for use of ozone therapy: [4]

• Pregnancy

• Glucose-6-phosphate-dehydrogenase deficiency (favism)

• Hyperthyroidism

• Severe anaemia

• Severe myasthenia

• Active hemorrhage

• Acute alcohol intoxication

• Recent Myocardial infarction
 
Discussion

Gingivitis and periodontitis are characterized by a local hypoxia of tissues and also by various microbic florae that may contain over 500 species. Accumulations of bacterial plaque in the gingival crevice area in an increased amount causes changes in the oral cavity ecology leading to both gingivitis and periodontitis. [33]

Dental biofilm makes it difficult for antibiotics in targeting putative periodontal pathogens. Higher concentrations of antibiotics are required to kill these organisms which are inevitably associated with toxic adverse effect on the host microbial flora. The application of ozone therapy in periodontics showed promising results. Both gaseous and aqueous ozone are used as a substitute to mechanical debridement. Ozone can be used to help treat periodontal disease by using ozonated water flushed below the gum line and/or ozone gas infiltrated into the gum tissue and supporting tissues.

Ozonated water (4mg/l) strongly inhibited the formation of dental plaque and reduced the number of sub gingival pathogens both gram positive and gram negative organisms. Gram negative bacteria, such as P. endodontalis and P. gingivalis were substantially more sensitive to ozonated water than gram positive oral streptococci and C. albicans in pure culture. [17] Furthermore ozonated water had strong bactericidal activity against bacteria in plaque biofilm. In addition, ozonated water inhibited the accumulation of experimental dental plaque in vitro. [17]

Dental researchers have started to examine the effects of ozonated fluids in periodontal disease. Huth et al in two papers in 2006 [19] and 2007 [26] examined the effect of ozone on periodontal tissues. The 2007 paper compared traditional periodontal anti-microbial products with the use of ozonated water. Both papers concluded that ozonated water has an excellent anti-microbial effect.

It resulted in toxic effect on human oral epithelial and fibroblast cells compared to antiseptics such as chlorehexidine digluconate, sodium hypochlorite and hydrogen peroxide during a 1-minute time period. [34,35] Ozone gas found to be toxic to the human oral epithelial and gingival fibroblast cells and aqueous ozone was more biocompatible than gaseous ozone. [19] The application of ozone therapy in chronic gingival and periodontal diseases, showed subjective and objective improvement of their status, as well as patients with periodontal abscess, with no exudation was observed. [36]

Brauner [37] has demonstrated that the combination of professional tooth cleaning and daily rinsing of the mouth with ozone water can improve clinical findings in cases of gingivitis and periodontitis. Plaque indices and a tendency to bleed, however, quickly return if the professional measures are interrupted. Rinsing the mouth with ozone water without any mechanical procedures for plaque reduction were unsuccessful.

There are many benefits to control oral hygiene and as a source of sterile water. However, patients should also be informed that there is an interaction of aqueous ozone with anti-microbials. This research has been published, illustrating the importance of potential interactions of dissolved ozone and prescribed anti-microbials. Patients who are taking a course of antibiotics may need to be informed that the use of ozonated water inactivates antibacterial agents [28] and in particular amoxicillin, [29] progesterone [30] and tetracycline. [31] What concerns the dentists is that ozone may inactivate the anti-microbial effects of triclosan. [32]

The effect of ozone on wounds obtained in the process of surgical and implantological procedures is used to prevent complications like after-surgical infection and to conduce to proper tissue healing. The use of ozone around implants is supported by published research showing that ozone not only effectively sterilizes the surfaces of both the implant and bone, but also switches on the reparative mechanisms allowing tissue regeneration around implant surfaces. [43] According to Matsumura et al [44,45] ozone does not have a major impact on stimulation of gingival cells for osteoblastic activity in the regeneration of the periodontium around implants.

Conclusions

Dentistry is changing as we are now using modern science to practice dentistry. In comparison with classical medicine modalities such as antibiotics and disinfectants, ozone therapy is quite inexpensive, predictable and conservative. The ozone therapy has been more beneficial than present conventional therapeutic modalities. This state of the art technology allows us to take a minimally invasive and conservative approach to dental treatment. The elucidation of molecular mechanisms of ozone further benefits practical application in dentistry. Treating patients with ozone therapy reduces the treatment time with a great deal of difference and it eliminates the bacterial count more precisely. The treatment is completely painless and increases the patients' acceptability and compliance with minimal adverse effects. Although more clinical research has to be done to standardize indications and treatment procedures of ozone therapy, still many different approaches are so promising, or already established, that hopefully the use of ozone therapy becomes a standard treatment for disinfection of an operation sites in dentistry.
 
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3276005/
J Nat Sci Biol Med. 2011 Jul-Dec; 2(2): 151–153.
doi:  10.4103/0976-9668.92318
PMCID: PMC3276005

Ozone therapy in dentistry: A strategic review

Rajiv Saini

Abstract

The oral cavity appears as an open ecosystem, with a dynamic balance between the entrance of microorganisms, colonization modalities, and host defenses aimed to their removal: To avoid elimination, bacteria need to adhere to either hard dental surfaces or epithelial surfaces. The oral biofilm formation and development, and the inside selection of specific microorganisms have been correlated with the most common oral pathologies, such as dental caries, periodontal disease, and peri-implantitis. The mechanical removal of the biofilm and adjunctive use of antibiotic disinfectants or various antibiotics have been the conventional methods for periodontal therapy. Ozone (O3) is a triatomic molecule, consisting of three oxygen atoms, and its application in medicine and dentistry has been indicated for the treatment of 260 different pathologies. The ozone therapy has been more beneficial than present conventional therapeutic modalities that follow a minimally invasive and conservative application to dental treatment. The exposition of molecular mechanisms of ozone further benefits practical function in dentistry.



http://www.ncbi.nlm.nih.gov/pubmed/18473030
J Contemp Dent Pract. 2008 May 1;9(4):75-84.

Ozone therapy in medicine and dentistry.

Nogales CG1 Ferrari PH, Kantorovich EO, Lage-Marques JL.

Abstract

AIM:

The purpose of this review is to present the potential for the incorporation of ozone therapy into the practice of dentistry.

BACKGROUND:

Ozone gas has a high oxidation potential and is 1.5 times greater than chloride when used as an antimicrobial agent against bacteria, viruses, fungi, and protozoa. It also has the capacity to stimulate blood circulation and the immune response. Such features justify the current interest in its application in medicine and dentistry and have been indicated for the treatment of 260 different pathologies. It can be used for the treatment of alveolitis as a replacement for antibiotic therapy, as a mouthwash for reducing the oral microflora, as well as the adherence of microorganisms to tooth surfaces. Ozone has been shown to stimulate remineralization of recent caries-affected teeth after a period of about six to eight weeks.

CONCLUSION:

The future of ozone therapy must focus on the establishment of safe and well-defined parameters in accordance with randomized, controlled trials to determine the precise indications and guidelines in order to treat various medical and dental pathologies. Scientific support, as suggested by demonstrated studies, for ozone therapy presents a potential for an atraumatic, biologically-based treatment for conditions encountered in dental practice.



http://www.ncbi.nlm.nih.gov/pubmed/16683682
Quintessence Int. 2006 May;37(5):353-9.

The use of ozone in dentistry and maxillofacial surgery: a review.

Stübinger S, Sader R, Filippi A.

Abstract

Ozone has been successfully used in medicine because of its microbiologic properties for more than 100 years. Its bactericide, virucide, and fungicide effects are based on its strong oxidation effect with the formation of free radicals as well as its direct destruction of almost all microorganisms. In addition, ozone has a therapeutic effect that facilitates wound healing and improves the supply of blood. For medical purposes, ozone may be applied as a gas or dissolved in water. Despite the advantages that the therapeutic use of ozone offers, reservations remain in terms of its application in the oral and maxillofacial area. Particularly, the gaseous application of ozone is critically evaluated because of its possible side effects on the respiratory system. The objective of this article is to provide an overview of the current applications of ozone in dentistry and oral surgery. Research was based on peer-reviewed sources found through a Medline/PubMed search and other textbooks, reviews, and journals.



http://www.ncbi.nlm.nih.gov/pubmed/19097384
Prague Med Rep. 2008;109(1):5-13.

Ozone and its usage in general medicine and dentistry. A review article.

Seidler V, Linetskiy I, Hubálková H, Stanková H, Smucler R, Mazánek J.

Abstract

Ozone, an allotropic form of oxygen, is successfully used in the treatment of different diseases for more than a hundred years. It is highly valued for various effects, such as antimicrobial, antihypoxic, analgesic, immunostimulating etc. on biological systems. These mechanisms of action supported with a lot of case reports and scientific studies allow using it in different fields of medicine. This review of literature is another attempt to summarize different modalities of ozone application in dentistry. Further studies are necessary to standardize indications and treatment protocols of this promising medical agent.



http://www.ncbi.nlm.nih.gov/pubmed/11093025
Caries Res. 2000 Nov-Dec;34(6):498-501.

Antimicrobial effect of a novel ozone- generating device on micro-organisms associated with primary root carious lesions in vitro.

Baysan A, Whiley RA, Lynch E.

Abstract

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.



http://www.ncbi.nlm.nih.gov/pubmed/14697022
Gerodontology. 2003 Dec;20(2):106-14.

Clinical reversal of root caries using ozone, double-blind, randomised, controlled 18-month trial.

Holmes J.

Abstract

OBJECTIVE:

To assess the effect of an ozone delivery system, combined with the daily use of a remineralising patient kit, on the clinical severity of non-cavitated leathery primary root carious lesions (PRCL's), in an older population group.

DESIGN:

A total of 89 subjects, (age range 60-82, mean +/- SD, 70.8 +/- 6 years), each with two leathery PRCL's, were recruited. The two lesions in each subject were randomly assigned for treatment with ozone or air, in a double-blind design, in a general dental practice. Subjects were recalled at three, six, 12 and 18 months. Lesions were clinically recorded at each visit as soft, leathery or hard, scored with a validated root caries severity index.

RESULTS:

There were no observed adverse events. After three months, in the ozone-treated group, 61 PRCL's (69%) had become hard and none had deteriorated, whilst in the control group, four PRCL's (4%) had become worse (p<0.01). At the six-month recall, in the ozone group, seven PRCL's (8%) remained leathery, the remaining 82 (92%) PRCL's had become hard, whilst in the control group, 10 PRCL's had become worse (11%) and one had become hard (p<0.01). At 12 and 18 months, 87 Subjects attended. In the ozone group at 12 months, two PRCL's remained leathery, compared to 85 (98%) that had hardened, whilst in the control group 21 (24%) of the PRCL's had progressed from leathery to soft, i.e. became worse, 65 PRCL's (75%) were still leathery, and one remained hard (p<0.01). At 18 months, 87 (100%) of ozone-treated PRCL's had arrested, whilst in the control group, 32 lesions (37%) of the PRCL's had worsened from leathery to soft (p<0.01), 54 (62%) PRCL's remained leathery and only one of the control PRCL's had reversed (p<0.01).

CONCLUSIONS:

Leathery non-cavitated primary root caries can be arrested non-operatively with ozone and remineralising products. This treatment regime is an effective alternative to conventional "drilling and filling".



http://www.ncbi.nlm.nih.gov/pubmed/15209994
Oral Microbiol Immunol. 2004 Aug;19(4):240-6.

Efficacy of ozone on survival and permeability of oral microorganisms.

Nagayoshi M, Fukuizumi T, Kitamura C, Yano J, Terashita M, Nishihara T.

Abstract

In the present study, we examined the effect of ozonated water on oral microorganisms and dental plaque. Almost no microorganisms were detected after being treated with ozonated water (4 mg/l) for 10 s. To estimate the ozonated water-treated Streptococcus mutans, bacterial cells were stained with LIVE/DEAD BacLight Bacterial Viability Kit. Fluorescence microscopic analysis revealed that S. mutans cells were killed instantaneously in ozonated water. Some breakage of ozonated water-treated S. mutans was found by electron microscopy. When the experimental dental plaque was exposed to ozonated water, the number of viable S. mutans remarkably decreased. Ozonated water strongly inhibited the accumulation of experimental dental plaque in vitro. After the dental plaque samples from human subjects were exposed to ozonated water in vitro, almost no viable bacterial cells were detected. These results suggest that ozonated water should be useful in reducing the infections caused by oral microorganisms in dental plaque.



http://www.ncbi.nlm.nih.gov/pubmed/16911107
Eur J Oral Sci. 2006 Aug;114(4):349-53.

Antibacterial effect of an ozone device and its comparison with two dentin-bonding systems.

Polydorou O, Pelz K, Hahn P.

Abstract

Microorganisms remaining beneath restorations can cause secondary caries and pulp damage. Because of this, antimicrobial treatment could be useful. The aim of this study was to evaluate the antibacterial effect of the HealOzone device on Streptococcus mutans and to compare it with the already proven activity of two dentin-bonding systems. Thirty-five human molars were divided into 5 groups. Cavities were then cut into the teeth (n = 28 cavities per group). After sterilization, the teeth were left in broth cultures of 10(6) colony-forming units (CFU) ml(-1) of S. mutans at 36 degrees C for 48 h. The appropriate treatment followed (group A, control; group B, Clearfil SE Bond; group C, Clearfil Protect Bond; group D, 40 s of treatment with ozone; and group E, 80 s of treatment with ozone), and the cavities were then filled with composite resin. After 72 h, the restorations were removed, dentin chips were collected with an excavator, and the total number of microorganisms was determined. All treatments significantly reduced the number of S. mutans present compared with the control group. The antimicrobial effect of both bonding systems and treatment with 80 s of ozone was significantly higher than the 40 s ozone treatment. In conclusion, HealOzone and the bonding systems show striking antimicrobial effects against S. mutans.



http://www.ncbi.nlm.nih.gov/pubmed/18280954
Oral Surg Oral Med Oral Pathol Oral Radiol Endod. 2008 Mar;105(3):e85-91. doi: 10.1016/j.tripleo.2007.10.006.

Effectiveness of ozonated water on Candida albicans, Enterococcus faecalis, and endotoxins in root canals.

Cardoso MG, de Oliveira LD, Koga-Ito CY, Jorge AO.

Abstract

The aim of this study was to evaluate the effectiveness of ozonated water in the elimination of Candida albicans, Enterococcus faecalis, and endotoxins from root canals. Twenty-four single-rooted human teeth were inoculated with C. albicans and E. faecalis, and 24 specimens were inoculated with Escherichia coli endotoxin. Ozonated water (experimental group) or physiologic solution (control group) was used as irrigant agent. Antimicrobial effectiveness was evaluated by the reduction of microbial counts. Lipopolissacharide complex presence was assessed by limulus amebocyte lysate test and B-lymphocyte stimulation. Data were analyzed by Wilcoxon and Mann-Whitney tests (5%). Ozonated water significantly reduced the number of C. albicans and E. faecalis at the immediate sampling, but increased values were detected after 7 days. Ozonated water did not neutralize endotoxin. It could be concluded that ozonated water was effective against C. albicans and E. faecalis but showed no residual effect. No activity on endotoxin was observed.



http://www.ncbi.nlm.nih.gov/pubmed/18618038
Quintessence Int. 2008 Mar;39(3):231-6.

The disinfecting effect of ozonized oxygen in an infected root canal: an in vitro study.

Stoll R, Venne L, Jablonski-Momeni A, Mutters R, Stachniss V.

Abstract

OBJECTIVES:

To determine the disinfecting effect of ozonized oxygen (120 seconds from the HealOzone generator, KaVo) on Enterococcus faecalis, representing bacteria that are difficult to eliminate in the root canals of human teeth, and to compare it with the conventional irrigants: sterile physiologic sodium chloride solution (negative control group), 3% hydrogen peroxide solution, 0.2% chlorhexidine solution, 1.5% sodium hypochlorite solution, and 3% sodium hypochlorite solution (positive control group).

METHOD AND MATERIALS:

The roots (n = 10 in each group) were sterilized, contaminated with the test microorganisms in a quantitative preparation, rinsed with the test solutions, and dried. The residual concentration of E faecalis was determined through another irrigation stage with the sodium chloride solution.

RESULTS:

The positive control group showed a significantly lower concentration of microorganisms than all the other groups, whereas the negative control group showed a significantly higher concentration compared to the other groups. The test groups showed low concentrations.

CONCLUSION:

Ozonized oxygen appears to be suitable for disinfecting root canal systems in cases where sodium hypochlorite is not indicated.



http://www.ncbi.nlm.nih.gov/pubmed/17026511
Eur J Oral Sci. 2006 Oct;114(5):435-40.

Effect of ozone on oral cells compared with established antimicrobials.

Huth KC, Jakob FM, Saugel B, Cappello C, Paschos E, Hollweck R, Hickel R, Brand K.

Abstract

Ozone has been proposed as an alternative antiseptic agent in dentistry based on reports of its antimicrobial effects in both gaseous and aqueous forms. This study investigated whether gaseous ozone (4 x 10(6) microg m(-3)) and aqueous ozone (1.25-20 microg ml(-1)) exert any cytotoxic effects on human oral epithelial (BHY) cells and gingival fibroblast (HGF-1) cells compared with established antiseptics [chlorhexidine digluconate (CHX) 2%, 0.2%; sodium hypochlorite (NaOCl) 5.25%, 2.25%; hydrogen peroxide (H(2)O(2)) 3%], over a time of 1 min, and compared with the antibiotic, metronidazole, over 24 h. Cell counts, metabolic activity, Sp-1 binding, actin levels, and apoptosis were evaluated. Ozone gas was found to have toxic effects on both cell types. Essentially no cytotoxic signs were observed for aqueous ozone. CHX (2%, 0.2%) was highly toxic to BHY cells, and slightly (2%) and non-toxic (0.2%) to HGF-1 cells. NaOCl and H(2)O(2) resulted in markedly reduced cell viability (BHY, HGF-1), whereas metronidazole displayed mild toxicity only to BHY cells. Taken together, aqueous ozone revealed the highest level of biocompatibility of the tested antiseptics.



http://www.ncbi.nlm.nih.gov/pubmed/19426912
Oral Surg Oral Med Oral Pathol Oral Radiol Endod. 2009 May;107(5):e73-9. doi: 10.1016/j.tripleo.2009.01.048.

Bactericidal effect of KTP laser irradiation against Enterococcus faecalis compared with gaseous ozone: an ex vivo study.

Kustarci A, Sümer Z, Altunbas D, Kosum S.

Abstract

OBJECTIVE:

The objective of this study was to evaluate the antimicrobial activity of potassium-titanyl-phosphate (KTP) laser and gaseous ozone in experimentally infected root canals.

STUDY DESIGN:

Eighty single-rooted teeth with straight canals were selected. After preparation and sterilization, the specimens were inoculated with 10 microL Enterococcus faecalis for 24 hours at 37 degrees C. The contaminated roots were divided into 2 experimental groups, 1 negative control group, and 1 positive control group of 20 teeth each: Group 1, KTP laser group; Group 2, gaseous ozone group; Group 3, sodium hypochlorite group (NaOCl) (negative control); and Group 4, saline group (positive control). Sterile paper points used to sample bacteria from the root canals were transferred to tubes containing 5 mL of brain heart infusion broth. Then 10-microL suspension was inoculated onto blood agar plates. The colonies of bacteria were counted and data were analyzed statistically using Kruskal-Wallis 1-way analysis of variance and Mann-Whitney U tests.

RESULTS:

There were statistically significant differences between all groups (P < .05). The saline group had the highest number of remaining microorganisms. Complete sterilization was achieved in the 2.5% NaOCl group. The KTP laser and gaseous ozone did not completely sterilize the root canals.

CONCLUSION:

Both KTP laser and gaseous ozone have a significant antibacterial effect on infected root canals, with the gaseous ozone being more effective than the KTP laser. However, 2.5% NaOCl was superior in its antimicrobial abilities compared with KTP laser and gaseous ozone.



http://www.ncbi.nlm.nih.gov/pubmed/22414842
J Endod. 2012 Apr;38(4):523-6. doi: 10.1016/j.joen.2011.12.020. Epub 2012 Feb 2.

Treatment of root canal biofilms of Enterococcus faecalis with ozone gas and passive ultrasound activation.

Case PD, Bird PS, Kahler WA, George R, Walsh LJ.

Abstract

INTRODUCTION:

Biofilms of resistant species such as Enterococcus faecalis pose a major challenge in the treatment of root canals with established periapical disease. This study examined the effects of gaseous ozone delivered into saline on biofilms of E. faecalis in root canals of extracted teeth with and without the use of passive ultrasonic agitation.

METHODS:

Biofilms of E. faecalis were established over 14 days in 70 single roots that had undergone biomechanical preparation followed by gamma irradiation. The presence and purity of biofilms were confirmed using scanning electron microscopy and culture. Biofilms were treated with saline (negative control), 1% sodium hypochlorite for 120 seconds (positive control), ozone (140 ppm ozone in air at 2 L/min delivered into saline using a cannula for 120 seconds), saline with passive ultrasonic activation (70 kHz and 200 mW/cm(2) applied to an ISO 15 file held passively within the canal, for 120 seconds), and ozone followed immediately by ultrasonic agitation. After treatment, samples were taken from the biofilm and serially diluted for plate counting.

RESULTS:

Analysis revealed that 1% sodium hypochlorite was the most effective disinfecting agent followed by ozone combined with ultrasonic agitation, ozone alone, and finally ultrasonic alone.

CONCLUSIONS:

Although none of the treatment regimes were able to reliably render canals sterile under the conditions used, ozone gas delivered into irrigating fluids in the root canal may be useful as an adjunct for endodontic disinfection.



http://www.ncbi.nlm.nih.gov/pubmed/15892361
J Adhes Dent. 2005 Spring;7(1):29-32.

Effect of ozone on enamel and dentin bond strength.

Schmidlin PR, Zimmermann J, Bindl A.

Abstract

PURPOSE:

To evaluate the influence of direct high-dose gaseous ozone application (2100 ppm) on dentin and enamel shear bond strength.

MATERIALS AND METHODS:

Ten bovine enamel and dentin samples per group were pretreated as follows: (I) ozone application (Healozone, KaVo) for 60 s alone or (II) with subsequent application of a fluoride- and xylitol-containing antioxidant (liquid reductant), (III) light-activated bleaching with 35% hydrogen peroxide for 5 min serving as negative control (Hi-Lite, Shofu), and (IV) untreated enamel and dentin (positive control). Specimens were bonded with a functional 3-step adhesive system (Syntac Classic, Ivoclar Vivadent) and restored with a composite (Tetric Ceram, Ivoclar Vivadent) according to the Ultradent method. After storage in water at 37 degrees C for 24 h, shear bond strength was measured using a Zwick universal testing machine. Data were analyzed using ANOVA and Scheffe's post hoc analysis.

RESULTS:

In concordance with the existing literature, bleaching resulted in significantly decreased bond strength (p < 0.05) on enamel specimens. No decrease in shear bond strength was detected for ozone-pretreated specimens compared to untreated controls.

CONCLUSION:

Despite a possible retention of surface and subsurface oxide-related substances during high-dose ozone application, shear bond strength was not impaired. Thus, adhesive restoration placement should be possible immediately after ozone application for cavity disinfection.



http://www.ncbi.nlm.nih.gov/pubmed/19701509
J Adhes Dent. 2009 Aug;11(4):287-92.

Enamel and dentin bond strength following gaseous ozone application.

Cadenaro M, Delise C, Antoniollo F, Navarra OC, Di Lenarda R, Breschi L.

Abstract

PURPOSE:

To evaluate the effects of gaseous ozone application on enamel and dentin bond strength produced by two self-etching adhesive systems.

MATERIALS AND METHODS:

The shear bond strength test was conducted to assess adhesion on enamel (protocol 1), while the microtensile bond strength test was performed on dentin (protocol 2). Protocol 1: 96 bovine incisors were randomly divided into 4 groups, and enamel surfaces were bonded in accordance with the following treatments: (1E) ozone + Clearfil Protect Bond; (2E) Clearfil Protect Bond (control); (3E) ozone + Xeno III; (4E) Xeno III (control). Ozone gas was applied for 80 s. Shear bond strength was measured with a universal testing machine. Protocol 2: 40 noncarious human molars were selected. Middle/deep dentin was exposed and bonded in accordance with the following treatments: (1D) ozone+Clearfil Protect Bond; (2D) Clearfil Protect Bond (control); (3D) ozone+Xeno III (4D) Xeno III (control). Four-mm-thick buildups were built on the adhesives, then specimens were sectioned in accordance with the nontrimming technique. Specimens were stressed until failure occurred, and failure modes were analyzed. Shear bond and microtensile bond strength data were analyzed using two-way ANOVA and Tukey's post-hoc test.

RESULTS:

No statistical differences were found between ozone treated specimens and controls, neither on enamel nor on dentin irrespective of the tested adhesive. Clearfil Protect Bond showed higher bond strength to enamel than Xeno III, irrespective of the ozone treatment (p < 0.05).

CONCLUSION:

The use of ozone gas to disinfect the cavity before placing a restoration had no influence on immediate enamel and dentin bond strength.



http://www.ncbi.nlm.nih.gov/pubmed/21417122
J Clin Pediatr Dent. 2010 Winter;35(2):187-90.

Effect of ozone pretreatment on the microleakage of pit and fissure sealants.

Cehreli SB, Yalcinkaya Z, Guven-Polat G, Cehreli ZC.

Abstract

OBJECTIVE:

This study investigated the effect of ozone pretreatment on the microleakage and marginal integrity of pit and fissure sealants placed with or without a self-etch 6th generation adhesive.

STUDY DESIGN:

Freshly-extracted, human third molars were randomly assigned into two main groups (n = 48): Group A: Fissures were pretreated with ozone; Group B: Fissures were left untreated. The teeth were further randomly divided into two subgroups (n = 24/each) so that half of teeth were sealed with a conventional fissure sealant (Fissurit F, Voco, Germany), while the remaining half received the same sealant bonded with a self-etch adhesive (Clearfil Protect Bond, Kuraray, Japan). Following thermal cycling (1000X), the specimens were subjected to dye penetration within 0.5% basic fuchsin for 24h. The extent of dye penetration was measured by image analysis. Kruskal Wallis and Mann-Whitney U tests were used for statistical analysis of the data (p = 0.05). Two randomly-selected sections from each group were observed under SEM RESULTS: In all groups, ozone pretreatment significantly reduced the extent of microleakage (p < 0.001). SEM investigation demonstrated better adaptation of the sealants in ozone-pretreated groups. Clearfil Protect Bond did not improve the marginal seal of Fissurit F (p > 0.05).

CONCLUSION:

Ozone pretreatment favorably affected the marginal sealing ability of the tested fissure sealants.