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Year : 2019  |  Volume : 22  |  Issue : 12  |  Page : 1758-1764

Do ozonated water and boric acid affect the bond strength to dentin in different adhesive systems?

1 Department of Restorative Dentistry, Faculty of Dentistry, Cumhuriyet University, Sivas, Turkey
2 Department of Restorative Dentistry, Faculty of Dentistry, Sakarya Univercity, Sakarya, Turkey

Date of Submission24-May-2019
Date of Acceptance08-Aug-2019
Date of Web Publication3-Dec-2019

Correspondence Address:
Dr. E Akturk
Department of Restorative Dentistry, Faculty of Dentistry, Cumhuriyet University, Sivas - 58140
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Source of Support: None, Conflict of Interest: None

DOI: 10.4103/njcp.njcp_281_19

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Aim: The aim of this in vitro study was to compare the effects of the application of three different cavity disinfecting agents to dentin on the micro-shear bond strength (μ-SBS) of one self-etch and two universal adhesive systems. Materials and Methods: In total, 120 caries-free human permanent molar teeth were used in this study. Mid-coronal dentin surfaces were revealed by cutting occlusal enamel and a standard smear layer was obtained by using 600-800-1200 grid silicon carbide abrasive papers. Specimens were randomly assigned to four groups according to the disinfectant used: Group 1: Control (no disinfectant); Group 2: 2% chlorhexidine based (Consepsis); Group 3: 10 ppm ozonated water (TeknO3zone); Group 4: 5% boric acid (Handmade). Each group was divided into three subgroups according to the type of adhesive (Clearfil SE Bond, OptiBond XTR, and Tokuyama Universal). Specimens were bonded using either Clearfil SE Bond, OptiBond XTR or Tokuyama Universal, which were employed according to the manufacturer's instructions. Resin composite microcylinders were bonded using Tygon® tubes for μ-SBS testing. After specimens were stored for 24 h, at 37°C in distilled water, μ-SBS test was measured with a universal test machine (LF Plus, Lloyd, Instrument). μ-SBS results were analyzed by one-way analysis of variance (ANOVA) and Tukey's tests. Results: When the mean microshear bond strength values of the control group were compared, the difference between the subgroups was not significant (P < 0.05). When the mean microshear bond strength values of the chx, ozonated water, and boric acid were compared, the difference between Clearfil SE Bond and Tokuyama Universal was significant (P < 0.05) and the difference between the other groups was not significant (P > 0.05). Conclusion: Ozonated water and boric acid may be as an alternative to other materials used as cavity disinfectants.

Keywords: Adhesive systems, boric acid, ozonated water

How to cite this article:
Akturk E, Bektas O O, Ozkanoglu S, G Akin E G. Do ozonated water and boric acid affect the bond strength to dentin in different adhesive systems?. Niger J Clin Pract 2019;22:1758-64

How to cite this URL:
Akturk E, Bektas O O, Ozkanoglu S, G Akin E G. Do ozonated water and boric acid affect the bond strength to dentin in different adhesive systems?. Niger J Clin Pract [serial online] 2019 [cited 2023 Jan 26];22:1758-64. Available from:

   Introduction Top

During tooth cavity preparation, the success of restorative treatment can be affected by bacterial remnants in the cavity walls, smear layer and dentin tubules. Bacteria remaining in the dentin tissue under a restoration may cause recurrent caries,[1] pulpal irritation[2],[3] and postoperative sensitivity.[4] For these reasons, the elimination of the bacteria from the cavity surfaces is of major importance.

To remove all the bacteria from the cavity preparation and to reduce the potential for residual caries, use of antibacterial solutions has been suggested in addition to the physical removal of carious dentin for the disinfection of dentinal cavities.[5],[6] Disinfectant solutions, such as chlorhexidine digluconate (CHX); sodium hypochlorite (NaOCl); disodium ethylenediamine tetraacetic acid (EDTA) dehydrate; and hydrogen peroxide (H2O2) are commonly used in modern-day settings to reduce or eliminate bacteria from cavity preparations.[7],[8] Alternatively, laser, ozone and boric acid can also be used to eliminate or reduce bacteria from the cavity.[9]

Chlorhexidine digluconate has been documented to have high antibacterial activity against both Gram-positive, especially Streptococcus mutans, and Gram-negative bacteria.[10],[11] Chlorhexidine digluconate in the form of 2% aqueous solution has been considered as a biocompatible[12],[13] and toxicologically safe disinfectant.[13],[14] It has been reported that the 2% solution is the most widely used CHX form in clinical dentistry and dental research.[15]

For many years, ozone is widely used in medicine because of its strong anti-oxidant effect, and a powerful antimicrobial agent is also used in dentistry. Fagrell et al.[16] reported a limited effect on bacterial growth for 5- to 10-s applications, but 60-s treatment was able to eliminate bacterial growth completely.

The element Boron is a bioactive trace element[17] which has metal and nonmetal properties and generally used in the weak inorganic acid form as boric acid, as a bactericide, fungicide and antiseptic.[18] There are research studies which use the antibacterial activity of different concentrations of boric acid.[19],[20]

The ideal dentin disinfectant should combine the possession of a potent antimicrobial action and should not interfere with the bonding of the subsequently applied adhesive system.[7] The advent of self-etching priming adhesives has simplified the use of resin-based bonding systems in clinical practice. Self-etching systems act by dissolving the smear layer but leave remnants of the smear plugs and etch the superficial enamel and dentin at the same time. It is this simultaneous etch and infiltration of the resin that is believed to be conducive to forming a well-infiltrated hybrid layer, even to dentin approximating the pulp.[21] A new family of adhesive systems, with chemical adhesion potential, which they are called multi-mode or universal adhesives due to their versatile instructions for use, have been launched. According to the respective manufacturers, universal adhesives can be used under the etch-and-rinse mode, the self-etch mode or with enamel selective etching.[22] The chemical interaction is a crucial characteristic of universal adhesives to enhance the durability of dentin–resin interfaces. Bond strengths are usually lower than those of two- and three-step etch-and-rinse adhesives but in line with results reported for self-etch adhesives.[23]

The aim of this in vitro study was to evaluate the bond strength of in tooth dentin of one self-etch adhesive system (Clearfil SE Bond, Kuraray) and two different universal adhesive systems (Tokuyama Universal, Tokuyama Dental; OptiBond XTR, Kerr) after disinfection with chlorhexidine gluconate, ozonated water or boric acid.

The null hypothesis examined the following:

  1. there is no difference in the μ-SBS between the self-etch and universal adhesives when bonded to superficial dentin pretreated with various dentin disinfectants;
  2. there is no difference in the μ-SBS in the control group between Clearfil SE Bond, OptiBond XTR and Tokuyama Universal groups; and
  3. boric acid has an adverse effect on micro shear bond strength.

   Materials and Methods Top


This study evaluated the effect of three dentin disinfectants: 2% CHX gluconate based (Consepsis Scrub, CHX); 10 ppm ozonated water (TeknO3 zone) and 5% boric acid (Handmade) on the μ-SBS of self-etch (Clearfil SE Bond and OptiBond XTR) and universal (Tokuyama Universal) adhesive systems. Materials composition and manufacturer presented in [Table 1].
Table 1: Materials used, composition and manufacturer

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Specimen preparation

In total, 120 caries-free human permanent molar teeth extracted for periodontal and orthodontic reasons were used in this study. The teeth were cleaned of calculus and soft tissue debris with a periodontal probe. The teeth were sectioned with a low-speed diamond disk saw (Isomet, Buehler Ltd., Lake Bluff IL, USA) under water coolant to expose mid-coronal dentin. The sections of the teeth including the roots were embedded in autopolymerizing acrylic resin to form cylinders 2 cm in diameter and 3 cm high. Dentin surfaces were flattened using 600–800 and 1200 grid silicon carbide abrasive paper under water coolant for 60 s to standardized the smear layer.

The specimens were randomly assigned to four groups according to the disinfectant used:

Group 1: No disinfectant (control)

Group 2: Chlorhexidine glukonate (CHX)

Group 3: Ozonated water (OW)

Group 4: Boric acid (BA)

In group 1, the specimens were not treated with any cavity disinfectant and served as control. The teeth in experimental groups were treated with one of the following cavity disinfectants: 2% CHX gel, 10 ppm ozonated water or 5% boric acid for 60 seconds. The dentin surfaces of the teeth were then dried with air for 10 s.


Each group was then randomly divided into three subgroups of ten teeth each according to the bonding agent used, Clearfil SE Bond, OptiBond XTR or Tokuyama Universal.

For Clearfil SE Bond, apply primer, scrub the dentin surface with a brushing motion for 20 s; gently air dry; apply adhesive with light brushing motion; air thin for 5 s; light cure 10 s (VALO Ultradent, USA).

For OptiBond XTR; apply primer; scrub the dentin surface with a brushing motion for 20 s; air thin for 5 s with medium air pressure, apply adhesive with light brushing motion for 15 s; air thin for 5 s; light cure 10 s (VALO Ultradent, USA).

For Tokuyama Universal, mix by dropping one drop of bottles A and B and apply to scrub the dentin surface with agitation for 20 s; air thin for 5 s.

After the completion of the adhesive application, a hybrid restorative composite (Estelite ∑ Quick Shade A1, Tokuyama Dental, Japan) was carefully inserted into the surface by packing the material into Tygon® tubing (Norton Performance Plastic Co., Cleveland, USA) with an internal diameter of 0.7 mm and a height of 0.5 mm and then light-cured for 20 s. After polymerization, Tygon tubes were removed gently by a scalpel.

Micro-shear bond strength (μ-SBS) testing

After storage in distilled water for 24 h, their μ-SBSs were measured using a universal testing machine (LF Plus, Lloyd, Instrument, Ametek Inc, England). A wire of 0.2 mm diameter was looped around the resin composite cylinder, making contact through half its circumference and was gently held flush against the resin–dentin interface. Shear force was applied to each specimen at a cross-head speed of 0.5 mm/min until failure occurred. The micro shear bond strength of composite resin to dentin was recorded in Newtons (N) and calculated in MPa taking into account the cross-sectional area of the composite buildup.

Statistical analysis

After recording the data, the results were subjected to statistical analysis using the software Statistical Packages for Social Sciences for Windows 22.0 (SPSS, Inc., Chicago, IL, USA). The Kolmogorov–Smirnov normality test was applied to the bond strength data. Because the data showed a normal distribution, the means of each group were analyzed by one-way ANOVA. Multiple comparisons were made by Tukey's test. P values less than 0.05 are considered to be statistically significant in all tests.

After measuring the bond strength, each specimen was examined visually using a binocular stereomicroscope (SMZ 800, Nikon, Japan) at × 25 magnification to determine the fracture modes. The percentage of the failure modes was calculated.

   Results Top

The mean and standard deviations of microshear bond strength for each adhesive systems and each groups are presented in [Table 2].
Table 2: Microshear bond strength

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The present study included four test groups: a control group, teeth that had been cleaned with a 2% chlorhexidine gel, teeth that had been cleaned with 10 ppm ozonated water and teeth that had been cleaned with 5%boric acid. No statistically significant difference in the bond strength between the resin used on the teeth in the OW and CHX groups, although the bond strength values were slightly higher in the CHX group. However, the bond strength of the control and BA groups were significantly lower than the bond strength in the CHX groups.

When the mean microshear bond strength values of the control group were compared, the difference between the subgroups was not significant (P < 0.05). When the mean microshear bond strength values of the chx, ozonated water and boric acid were compared, the difference between Clearfil SE Bond and Tokuyama Universal was significant (P< 0.05) and the difference between the other groups was not significant (P > 0.05). Therefore, the results of this in vitro study support the rejection of the null hypothesis that there is no difference in the μ-SBS between the self-etch and universal adhesives when bonded to superficial dentin pretreated with various dentin disinfectants.

In general, the use of disinfectants increased the bond strength when Clearfil SE Bond and OptiBond XTR were used compared with the control specimens. CHX prior to the use of Clearfil SE bond and OptiBond XTR recorded the highest mean bond strength followed by the ozonated water and boric acid.

The percentages of the fracture modes observed with all tested groups are presented in [Table 3]. The predominant mode of failure with all tested groups was adhesive.
Table 3: Fracture modes of the all test groups

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   Discussion Top

In the present study assessment the effect of the application of three different disinfecting agents on bond strength of an self-etch and universal adhesive systems was carried out using μ-SBS testing. Micro-shear test generates less stress or damage during the preparation of specimens compared to the micro-tensile alternative.[24]

In the present study, OptiBond XTR (universal adhesive system) and Tokuyama Universal (universal adhesive system) tested in the control group mean bond strength was not significantly different from that of Clearfil SE Bond, a proven standard for self-etch primer systems. These findings correlate with the results of Walter et al.,[25] who reported that no significant difference in the bond strength was observed between Clearfil SE Bond and OptiBond XTR. Therefore, the second null hypothesis was accepted.

Tokuyama Universal is a two-component, one-stage universal adhesive system that is chemically polymerized. There is not a sufficient number of studies in the literature regarding the bonding strength of Tokuyama Universal to dentin. The application of disinfectants after cavity preparation and before tooth restoration is gaining acceptance as they are considered to eliminate potential risks due to bacterial activity without having an adverse effect on the bonding performance of adhesive systems.

One antibacterial agent that is commonly used in dental practice is the CHX. CHX is among the well-known antimicrobial agents and matrix metalloproteinase inhibitors.[5],[26] Many authors expected that CHX can improve dentin bond strength while exerting antibacterial effects but the results are controversial. CHX has an excellent rewetting capacity and a strong affinity to tooth structure which is thought to improve the bond strengths of the adhesive to dentin.[27] The results of the present study revealed that CHX pretreatment enhanced the bonding performance of the self-etch adhesive examined. This finding is in line with the study by Pilo et al.,[27] and Mohammed Hassan et al.,[28] who reported that pretreating the dentin surface with CHX improved the bond strength of self-etch adhesive systems.

Some authors reported no statistically significant differences in the bond strength with self-etch adhesive systems and after 2% CHX application.[29],[30],[31] Similar results have been reported by Mobarak and coworkers,[29],[31] who found no adverse effects of CHX pretreatment on microshear bond strength (μSBS) of dentin bonded with a self-etch bonding system (Clearfil SE Bond).

However, Ercan et al.[32] and Sharma et al.[33] reported that chlorhexidine solution negatively affected the bond strength of Clearfil SE Bond. The adverse effect on bond strength of 2% CHX solutions associated with self-etch bonding systems and cement may be explained by the presence of functional monomer, 10-methacryloyloxydecyl dihydrogenphosphate (MDP), in the bonding resin of self-etch adhesive systems, which might have been affected by CHX bonding.[34] It is possible that the lack of consistency in these research results could be attributed to differences in the methodology, dentin substrate and curing unit employed.

Ozone is known to be a strong oxidizer. Hence, it possesses antibacterial activities by disrupting the cell wall and cytoplasmic membrane of bacteria and, therefore, destruction of the microorganism.[35] In dental applications, O3 can be used in one of three forms: Gaseous, water, or oil.[36] There is not enough evidence about OW's effect on bond strength. In the present study, we found that the use of OW significantly increased μ-SBS values. As such, the findings of the current study were contrary to the outcomes of some previous studies. For example, Dalkiliç et al.[5] investigated the effect of different disinfecting methods of self-etch adhesive systems on the dentin initial bond strength and reported that ozone reduced bond strength. Pithon et al.[37] and Hubbezoǧlu et al.[38] reported that application of OW did not affect bond strength.

However, Oznurhan et al.[39] found that the use of OW (3–4 ppm) significantly increased μTBS values. It is possible that the lack of consistency in the results of the present study could be attributed to differences in the methodology, different concentration ozonated water and dentin substrate employed.

Although there are some studies on the effects of different antibacterial agents in dentistry and medical applications, a trend towards studies evaluating the effectiveness of boron or boric acid is observed. Boric acid, which has antibacterial activity, is frequently used in medical applications as an inexpensive treatment agent in the treatment of some infections.[40],[41] In a study that investigated the effects of 0.4% to 5% concentrations of boric acid on Candida albicans, it was reported that it showed antifungal activity and both inhibited clinical isolates and killed 50% to 90%.[22] In another study in which 12% solution of boric acid was examined, Stapylococcus aureus, Streptococcus mutans, Enterococcus faecalis,  Pseudomonas aeruginosa Scientific Name Search terococcus faecium (Vancomycin resistant), Klebsiella pneumonia,  Escherichia More Details coli such as gram (+) and gram (-) have been reported to be effective on bacteria.[23] Considering these studies, it may be considered as an alternative to other materials used as cavity disinfectants.

In this study, 5% boric acid solution was applied as a cavity disinfectant before restoration. The results of the current study indicate that boric acid does not have an impact on the bond strength of composite resin, regardless of the type of bonding system employed (self-etching). This finding is in line with the study by Ercan et al.[42] who reported that pretreating the dentin surface with boric acid did not affect the bond strength of self-etch adhesive systems. The fact that boric acid does not have any impact on bond strength can be attributed to the fact that its penetration into the dentin structure is limited. Therefore, the third null hypothesis was rejected.

Hence, the antibacterial effect of boron and its bond strength-friendly properties may make boric acid a viable alternative as a cavity disinfectant material. Although there have been studies of boric acid used at different concentrations, there is a need for further research to determine which concentration is most effective against cariogenic bacteria and whether or not it affects the bond strength and microleakage of restorations.

   Conclusion Top

Under the limitation of this in vitro study, it can be concluded that:

  1. In this study which researched alternative methods to traditional disinfectants, the results showed that ozonated water and boric acid could be used for disinfection before composite restorations. Further,in vivo studies with a long-term follow-up are necessary to compare the effectiveness of ozonated water and boric acid as cavity disinfectants.
  2. Optibond XTR (a two-step universal adhesive), Clearfil SE Bond (a two-step self-etch adhesive) and Tokuyama Universal (a two-step universal adhesive) yielded similar results. These results suggest that immediate bond strength are not related to the curing systems and number of steps characterizing the bonding systems but to their chemical formulations.

Financial support and sponsorship


Conflicts of interest

There are no conflicts of interest.

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  [Table 1], [Table 2], [Table 3]

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