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 Table of Contents  
ORIGINAL ARTICLE
Year : 2019  |  Volume : 13  |  Issue : 1  |  Page : 12-18

Effects of various agents and laser systems on antibacterial activity and microtensile bond strength when used for cavity disinfection


1 Department of Pediatric Dentistry, Faculty of Dentistry, Recep Tayyip Erdoğan University, Rize, Turkey
2 Department of Pediatric Dentistry, Faculty of Dentistry, Karadeniz Technical University, Trabzon, Turkey
3 Department of Microbiology, Faculty of Medicine, Karadeniz Technical University, Trabzon, Turkey
4 Department of Microbiology, Nevsehir Public Hospital, Nevsehir, Turkey

Date of Web Publication24-Jul-2019

Correspondence Address:
Dr. Ipek Arslan
Department of Pediatric Dentistry, Faculty of Dentistry, Recep Tayyip Erdoğan University, Rize, postal code 53100
Turkey
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/jdl.jdl_16_18

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  Abstract 

Context: Cavity disinfection is recommended with the routine caries removal methods in order to eliminate the microorganisms and to reduce potential secondary caries. Aims: The aims of this study were to evaluate the antibacterial effects of Corsodyl; Cervitec; Cervitec Plus; Fluor Protector agents and FotoSan; diode laser; and erbium, chromium: yttrium, scandium, gallium, garnet (Er, Cr:YSGG) laser systems on Streptococcus mutans and Lactobacillus acidophilus as well as their effects on the microtensile bond strength. Materials and Methods: A cavity tooth model test was used to determine antibacterial activity after which the effects of the same agents and systems on the microtensile bond strength were evaluated. Eight cylindrical cavities were prepared on the dentin surface of 24 bovine incisors, and 12 of them were left in contact with S. mutans; others were left in contact with L. acidophilus. Test agents and systems were applied, and standardized amounts of dentin chips were obtained from the cavity walls. The number of bacteria recovered was counted. The effect of tested agents and systems on bond strength was evaluated with microtensile bond strength test. Statistical Analysis Used: Statistical analysis was carried out using the Kruskal–Wallis and Mann–Whitney U tests for the cavity tooth model test, and one-way ANOVA and Tukey test for microtensile bond strength test. Results: Test results showed that all of the disinfection methods demonstrated significant antibacterial activity on both S. mutans and L. acidophilus (P < 0.01). The agents used in this study significantly reduced the microtensile bond strength (P < 0.05) whereas the Er, Cr:YSGG laser significantly increased the bond strength (P < 0.01). Conclusions: Er, Cr:YSGG laser can be recommended for cavity disinfection due to its superior antibacterial activity and increased bond strength.

Keywords: Antibacterial, cavity disinfection, laser, L. acidophilus, microtensile bond strength, S. mutans
Key Messages: Cavity disinfection is recommended to reduce potential secondary caries. Er, Cr:YSGG laser can be recommended for cavity disinfection due to its superior antibacterial activity and increased bond strength.


How to cite this article:
Arslan I, Baygin O, Bayram G, Akyol R, Tuzuner T. Effects of various agents and laser systems on antibacterial activity and microtensile bond strength when used for cavity disinfection. J Dent Lasers 2019;13:12-8

How to cite this URL:
Arslan I, Baygin O, Bayram G, Akyol R, Tuzuner T. Effects of various agents and laser systems on antibacterial activity and microtensile bond strength when used for cavity disinfection. J Dent Lasers [serial online] 2019 [cited 2019 Aug 19];13:12-8. Available from: http://www.jdentlasers.org/text.asp?2019/13/1/12/263343




  Introduction Top


Traditional restorative dentistry aims to remove all infected and affected tooth structures to prevent further cariogenic activity.[1] However, several studies have reported the existence of bacteria in dentin even with caries detector dye usage.[2],[3] For these reasons, cavity disinfection is recommended in order to eliminate the microorganisms and to reduce potential secondary caries, pulp sensitivity, and pulp inflammation.[4],[5]

For this reason, this study aimed to evaluate and compare the cavity disinfection efficacy of Corsodyl; Cervitec; Cervitec Plus; Fluor Protector agents and FotoSan; erbium, chromium: yttrium, scandium, gallium, garnet (Er, Cr:YSGG) laser, and diode laser systems as well as their effects on microtensile bond strength of compomer restorations.


  Materials and Methods Top


Twenty-four bovine incisors were collected for antibacterial activity test and 24 human deciduous molar teeth were collected for the microtensile bond strength test. Following the extraction, the teeth were cleaned with pumice and stored in 0.5% chloramine T aqueous. Ethical approval was obtained from the ethics committee of Karadeniz Technical University, Faculty of Medicine (Protocol # 2015/149).

Antibacterial activity test

The antibacterial activity test was performed according to the tooth cavity method modified by Özer et al.[6] Enamel of bovine teeth was eroded with a water-cooled diamond bur to obtain flat dentinal surfaces. Six cylindrical cavities were prepared on the buccal surfaces and two cylindrical cavities were prepared on the lingual surfaces of each tooth without pulp exposure. The cavities were 1mm in width and 2mm in depth. Then, the teeth were sterilized in an autoclave for 15 minutes at 121°C. The teeth were placed in brain heart infusion broth (Oxoid, England, UK) and incubated for 24h at 37°C, to confirm sterility. Afterwards, each tooth was transferred to 2mL of sterile physiologic saline in an individual tube, and stored for 24 hours at 37°C to wash out the culture medium to avoid dehydration. The teeth were dried with sterile paper points. The teeth were then randomly divided into two groups of 12 teeth each. The first group was contaminated with a broth culture of Streptococcus mutans ATCC 25175 (MediMark, Grenoble, France) and incubated at 37°C for 72 hours. The second group was contaminated with a broth culture of Lactobacillus acidophilus ATCC 4356 (MediMark, Grenoble, France). After incubation, the teeth were dried with sterile paper points and gentle air flow. Each cavity disinfection method was applied to one of the eight cavities according to the manufacturers’ instructions as follows.

Corsodyl group: A 1% CHX Digluconate gel (GlaxoSmithKline, Dallas, TX) was applied to the dentin for 1 minute according to the manufacturer’s instructions. Excess gel was removed from the cavity with a clean cotton pellet.

Cervitec group: A combination of 0.2% CHX Digluconate and Sodium Fluoride gel (Ivoclar Vivadent, Ellwangen, Germany) was applied to the dentin for 2 minutes according to the manufacturer’s instructions. Excess gel was removed from the cavity with a clean cotton pellet.

Cervitec Plus group: A combination of 1% CHX Diacetate and 1% thymol varnish (Ivoclar Vivadent) was applied to the dentin for 2 minutes according to the manufacturer’s instructions.

Fluor Protector group: A 1% difluorosilane varnish (Ivoclar Vivadent) was applied to dentin for 1 minute according to the manufacturer’s instructions.

FotoSan group: A photosensitizer containing 0.01% toluidine blue was applied to the dentin. The teeth were irradiated with red light (660nm wavelength and 100 mW output) for 30 seconds according to the manufacturer’s instructions.

Diode laser group: The dentin surfaces were irradiated with a diode laser (ezLase; Biolase Technology, San Clemente, CA) with a wavelengths of 940nm, 1-W power output, and 20-Hz frequency. A sapphire tip, 600 µm in diameter and 6mm in length was used to deliver the laser light. Irradiation time was 5 seconds, repeated five times with an interval time of 15 seconds.

Er, Cr:YSGG laser group: The dentin surfaces were irradiated with an Er, Cr:YSGG laser (Waterlase MD, Biolase Technology Inc., San Clemente, CA, USA) with a wavelength of 2780nm, 1-W power output, and 20-Hz frequency. A sapphire tip, 600 µm in diameter and 6mm in length, was used to deliver the laser light. Irradiation time was 5 seconds, repeated five times with an interval time of 15 seconds.

Control group (CG): No cavity disinfection methods were applied to the teeth.

After application of the disinfection methods, all of the cavities were sealed with a temporary restorative material (Cavit G; 3M ESPE, Seefeld, Germany). The teeth were kept separately in sterile physiological saline at 37°C for 72 hour. The teeth were then removed from the sterile physiological saline and kept in a freezer at −25°C for 1 hour for cooling. Standardized amounts of dentin chips (25 ± 5mg) were collected from the circumferential cavity walls (except pulpal floor) and placed into sterile  Petri dish More Detailses using carbide fissure crosscut burs. A new sterile bur was used for each cavity to prevent overheating of the dentinal walls during the cutting action. The suspension with dentin chips collected was diluted in 2mL sterile physiological saline, and serial dilutions of 10:1, 10:2, and 10:3 were obtained. The number of viable bacterial cells was determined by counting the colony-forming units (CFUs) on 5% sheep blood agar (Becton Dickinson GmbH, Heidelberg, Germany) plates for S. mutans and MRS agar for L. acidophilus. This analysis was repeated three times.

Statistical analyses were performed with SPSS 15.0 for Windows (SPSS Inc., Chicago, IL). Distribution of the data was evaluated with a Shapiro–Wilks test. Because the data were nonparametric, Kruskal–Wallis and Mann–Whitney U tests (P ˂ 0.05) were applied.

Microtensile bond strength test

The roots of the teeth were embedded in a self-cured acrylic resin in a square plastic mould. The occlusal surfaces of the all the teeth were removed with a low-speed diamond wheel saw (Micracut 125, Metkon Technology, Bursa, Turkey). After the occlusal surfaces were removed, abrasive waterproof paper (No: 600) was used to smooth the dentin surfaces. The teeth were divided into eight groups, and each group was disinfected with one of the protocols as described previously in the antibacterial activity test. No cavity disinfection methods were applied to the teeth of the CG.

The bonding agent (Prime & Bond NT; Dentsply, Konstanz, Germany) was applied to the dentin surfaces and air dried gently then light cured for 20 seconds. Approximately 5mm of thick compomer (Dyract Extra; Dentsply, Konstanz, Germany) was built up on three layers and each layer was light cured for 20 seconds.

The teeth were stored in distilled water for 24 hours and serially sectioned perpendicular to the compomer surfaces to form 1 × 1mm2 thick resin slabs with a low-speed diamond wheel saw. Three or four sticks were obtained from each tooth (n:10). The sticks were attached to the jig of the microtensile test machine (Bisco Inc, Schaumburg, IL) with cyanoacrylate glue and forced in tension at a crosshead speed of 0.5mm/min. The cross-sectional area at the site of failure was measured with a digital caliper (Mitutoyo, Tokyo, Japan). The microtensile bond strength value was derived in Newtons by dividing the enforced force at time of fracture by the bond area (mm2).

Distribution of the data was evaluated with a Shapiro–Wilks test. Because the data were parametric, one-way ANOVA and Tukey tests (P ˂ 0.05) were applied.


  Results Top


Antibacterial activity test

The tooth cavity model test results showed that all of the disinfection methods produced significant antibacterial activity in comparison to the CG against both S. mutans and L. acidophilus (P < 0.01).

Streptococcus mutans

The means of the number of S. mutans CFUs (cfu/mL) recovered from the cavity walls and standard deviations for all experimental groups are shown in [Figure 1].
Figure 1: Number of Streptococcus mutans recovered after the tooth cavity test. *Statistically different groups (P < 0.001)

Click here to view


Statistically significant differences were found between the FotoSan and Fluor Protector groups (P = 0.016) and between the Cervitec and Fluor Protector groups (P = 0.008).

Lactobacillus acidophilus

The means of the number of L. acidophilus CFUs (cfu/mL) recovered from the cavity walls and standard deviations for all experimental groups are shown in [Figure 2].
Figure 2: Number of Lactobacillus acidophilus recovered after the tooth cavity test. *Statistically different groups (P < 0.001)

Click here to view


Statistically significant differences were found between the Corsodyl and Fluor Protector groups (P = 0.013).

Microtensile bond strength test

The mean, standard deviation, and maximum and minimum microtensile bond strength values as well as the P value of the comparison with the CG are shown in [Figure 3].
Figure 3: The mean, standard deviation, and maximum and minimum microtensile bond strength values as well as the P value of the comparison with the control group. aGroups statistically different from control group (P < 0.01). bGroups statistically different from Er, Cr:YSGG laser group (P < 0.01). cGroups statistically different from diode group (P < 0.05). dGroups statistically different from FotoSan group (P < 0.05)

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There was no statistically significant differences between the CG and FotoSan group as well diode laser group (P > 0.05).

Er, Cr:YSGG group showed significantly greater microtensile bond strength value than the CG (P < 0.01). Also Corsodyl, Cervitec, Cervitec Plus, and Fluor Protector groups showed significantly lesser microtensile bond strength value than the CG (P < 0.05)


  Discussion Top


The presence of bacteria on restored tooth is the major cause of secondary caries and the failure of restoration.[7] None of the caries-removal methods used presently eliminate all of the microorganisms in the cavities.[8] Thus, cavity disinfection procedures are recommended to eliminate these residual bacteria.[4] Various cavity disinfectants are currently in use including CHX, fluoride gels, sodium hypochlorite, benzalkonium-based solutions, propolis, and aloe vera.[9],[10] Technological devices such as lasers and photo-activated disinfection (PAD) are alternative procedures that may be used for cavity disinfection.[11]

Many techniques have been used to investigate the antibacterial action of cavity disinfection. The cavity tooth model test, modified to use bovine incisors instead of human molars, was used by Türkün et al.[11] Bovine incisors provide more dentine surfaces allowing more cavities to be prepared on the same tooth. Agar well and disc diffusion techniques are the models used for antibacterial activity determination and comparison. The disadvantage of these methods is that the diffusion rate of antibacterial solution into hydrophilic agar can affect the results. For this reason, the cavity tooth model that simulated the conditions of clinic is preferred.[11]

Streptococcus mutans and L. acidophilus are the most important colonisers that are responsible for dental caries.[12] For this reason, this study evaluated and compared the cavity disinfection efficacy of Corsodyl; Cervitec; Cervitec Plus; Flour Protector agents and FotoSan; Er, Cr:YSGG; and diode laser systems.

CHX is accepted as the gold standard of antibacterial agents and is commonly studied as a cavity disinfectant.[13] The superior antibacterial effect of CHX when used as a cavity disinfectant has been proved many times.[11],[13] In this study, Corsodyl gel containing 1% CHX Digluconate, Cervitec gel (combination of 0.2% CHX Digluconate and sodium fluoride gel), and Cervitec Plus Varnish (combination of 1% CHX Diacetate and 1% Thymol) were used. All of the CHX forms had superior antibacterial effects. Only Cervitec showed significantly less antibacterial activity than Fluor Protector against S. mutans. On the contrary, Mohan et al.[14] found that 2% CHX gel showed a greater antibacterial effect than acidulated phosphate fluoride (APF) gel. This finding can be explained by the lower CHX content of Cervitec.

Fluoride is the most widely used anticaries agent in dentistry and its antibacterial activity has been demonstrated many times.[15],[16] In this study, Fluor Protector varnish, containing 1% difluorosilane, demonstrated the highest antibacterial effect against S. mutans and significant antibacterial effect against L. acidophilus. Pinar Erdem et al.[17] reported that despite the fact that Fluor Protector contains lower fluoride concentration than Bifluoride 12, its antibacterial effect was better than that of Bifluoride 12 against S. mutans biofilms. This finding was explained by the silane content of Fluor Protector. In this study, Fluor Protector showed a greater antibacterial effect than Cervitec, but Mohan et al.[14] reported that CHX gel had a higher antibacterial effect than APF gel. This finding may be also explained by the silane content of Fluor Protector.

PAD is one of the disinfection methods that can be used for cavity disinfection. In this study, FotoSan was used as PAD system and toluidine blue as photosensitizer. FotoSan exhibited significant antibacterial action against S. mutans and L. acidophilus. However, it showed significantly less antibacterial action than Fluor Protector against S. mutans. To the best our knowledge, there is no previous published report that compares the antibacterial activity of PAD and fluoride. In addition, various studies have reported antibacterial action of PAD.[18],[19],[20] Pereira et al.[20] reported the antibacterial action of PAD against S. mutans and Streptococcus sanguinis, and suggested the use of PAD in the control of caries and periodontal diseases.

Laser therapy is whether associated with a photosensitizer or not is effective against oral bacteria. Despite its well-known antibacterial action, however, the number of studies regarding the use of laser for cavity disinfection is limited.[11],[14] In this study, the diode laser exhibited significant antibacterial action against S. mutans and L. acidophilus. A previous study that compared the antibacterial action of CHX, APF gel, Brazilian propolis, and diode laser reported that CHX, Brazilian propolis, and diode laser were equally effective.[14] This result is consistent with the results of this study.

When compared to the extensive research into the antibacterial effects of different lasers including Er, Cr:YSGG laser, on endodontic and periodontal pathogens, much less attention has been given to pathogens associated with dental caries. To the best of our knowledge, only Türkün et al.[11] have investigated the antibacterial activity of Er, Cr:YSGG laser against S. mutans. The researchers compared the antibacterial effects of Er, Cr:YSGG laser with 0.75 and 1 W power outputs and CHX gluconate-based cavity disinfectant against S. mutans and found that the antibacterial activity on S. mutans demonstrated by Er, Cr:YSGG laser with both energy outputs was similar to that of the tested CHX gluconate-based cavity disinfectant. In this study, similar to the previous studies, Er, Cr:YSGG laser showed strong antibacterial activity, similar to that of CHX.

Although cavity disinfection is recommended to eliminate the microorganisms and to reduce potential secondary caries, pulp sensitivity, and pulp inflammation, when disinfectant is used on cavities, it is important that cavity disinfection protocols do not interfere with the bonding between the resin materials and teeth.[21] For this reason, in this study the effects of materials and systems, tested for antibacterial efficacy, were also evaluated for their effects on the microtensile bond strength of the compomer restorations.

The results regarding the effects of CHX on dentin bond strength are controversial. Although many publications have reported that CHX decreased the bonding of an adhesive to dentin,[22] other publications have reported no adverse effect on bond strength.[23] This disparity can be explained by the increasing affinity of CHX to the tooth structure with acidification. In addition, CHX increases the free surface energy of enamel and dentin. Therefore, CHX can bind easily to a phosphate group, and then increase the bonding structure with acidification.[22] In this study, Prime & Bond NT was used, which needed no acidification in primary tooth compomer restorations. Corsodyl, Cervitec, and Cervitec Plus groups, which contain CHX, decreased the bond strength similar to the result of the study by Kapdan and Oztas.[22]

The studies regarding the effect of fluoride on bonding strength are limited, and in the majority of the studies that exist, fluoride was used as a desensitizer. Akca et al.[24] evaluated the bond strength of resin composite to dentin following the application of various dentin desensitizers, one of which was Fluor Protector. The researchers found that pretreatment of dentin surfaces with Fluor Protector reduced the bond strength values of composite restorations. In this study, Fluor Protector decreased bond strength as well, consistent with the study of Akca et al.[24] In another study, the effect of fluoride varnish on the shear bond strength of brackets was investigated. The results of that study showed that fluoride varnish had no adverse effect on the shear bond strength of the brackets.[25] This controversial result can be explained by noting that this study investigated the strength of bonding to enamel, not to dentin.

Only a few reports that investigating the effect of PAD on the bond strength of adhesive restorations exist in the literature. Sekhar et al.[26] found that PAD decreased the bond strength of resin-modified glass ionomer cement. In this study, it was found that PAD had no adverse effect on microtensile bond strength. This controversial result can be explained by differences in restorative materials.

Laser irradiation on dental hard tissues has been widely studied in dentistry.[11],[14] However, studies that compare laser types are limited. Generally, studies have reported on the relation between the increased bond strength of adhesives and laser irradiation. In this study, the results of Er, Cr:YSGG laser group agree with those of other studies, that Er, Cr:YSGG laser irradiation increased the microtensile bond strength of adhesives.[23],[27] On the other hand, diode laser increased the microtensile bond strength of compomer restorations but the results are not statistically different (P = 0.218). This finding can be explained by noting that other studies used composite, whereas in this study, compomer was used.

Although the results of some groups in this study are encouraging, controlled clinical studies are necessary to find ideal cavity disinfection procedures.


  Conclusion Top


Under the limitations of this study, all of the tested materials and systems had antibacterial activity against S. mutans and L. acidophilus as cavity disinfectants. However, Corsodyl, Cervitec, Cervitec Plus, and Fluor Protector reduced the bond strength of compomer restorations. The findings from this study showed that FotoSan and diode laser may be used for cavity disinfection due to their antibacterial effects and lack of adverse effects on the microtensile bond strength of compomer restorations. In addition, Er, Cr:YSGG laser can be recommended for cavity disinfection due to its superior antibacterial activity and increased bond strength.

The results of this study are encouraging, and further studies are needed to determine the effects of the tested materials and systems under in vivo conditions.

Financial support and sponsorship

Nil.

Conflicts of interest

There are no conflicts of interest.



 
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