|Year : 2012 | Volume
| Issue : 4 | Page : 469-472
|In vitro antimicrobial activity of AH Plus, EndoREZ and Epiphany against microorganisms
Lilian Eiko Maekawa1, Maria Renata Giazzi Nassri2, Camila Kaori Ishikawa3, Carolina Martins3, Adriana Chung1, Cristiane Yumi Koga-Ito4
1 Department of Restorative Dentistry, School of Dentistry of São José dos Campos, São Paulo State University - UNESP, São José dos Campos, São Paulo, Brazil
2 Department of Endodontics, School of Dentistry of São Paulo University - USP, São Paulo, Brazil
3 Department of Endodontics, School of Dentistry of Mogi das Cruzes University - UMC, Mogi das Cruzes, São Paulo, Brazil
4 Department of Biosciences and Oral Diagnosis, School of Dentistry of São José dos Campos, São Paulo State University - UNESP, São José dos Campos, São Paulo, Brazil
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|Date of Web Publication||20-Dec-2012|
| Abstract|| |
Objective : The aim of the present study was to evaluate the antimicrobial activity of endodontic sealers against microorganisms.
Materials and Methods : The agar diffusion method was used. A double base layer of Mueller Hinton agar was done. The microorganisms used were: Candida albicans, Enterococcus faecalis, Escherichia coli and Staphylococcus aureus. The wells were obtained by removing a standardized portion of the agar. After the distribution of the sealers, Petri plates were incubated for 24 h. Inhibition halos formed around the wells were measured.
Results : Epiphany did not show any antimicrobial activity on the tested microorganisms (without inhibition halo). The AH Plus showed the greatest inhibition halo on C. albicans followed by EndoREZ on S. aureus. EndoREZ also showed greater inhibition halo in comparison to AH Plus on E. faecalis and E. coli.
Conclusion : It could be concluded that AH Plus and EndoREZ showed antimicrobial activity against all the tested microorganisms. No antimicrobial activity was observed for Epiphany.
Keywords: Agar diffusion method, endodontic sealers, microorganisms
|How to cite this article:|
Maekawa LE, Nassri MG, Ishikawa CK, Martins C, Chung A, Koga-Ito CY. In vitro antimicrobial activity of AH Plus, EndoREZ and Epiphany against microorganisms. Indian J Dent Res 2012;23:469-72
Cleaning and disinfection of root canals are essential steps for the successful endodontic treatment,  due to the role of the microorganisms and their byproducts in the induction of pulp lesions and periapical inflammatory reactions.  . Nevertheless, even after the biomechanical prepare and use of intracanal medication, microorganisms might remain inside the dentinal tubules and can proliferate inducing an inflammatory process.  Marginal microinfiltration may also contribute to the recontamination of the root canal. This way, the obturation of the root canal is essential to avoid the recontamination by microorganisms from the oral cavity and the remaining ones in the dentinal tubules. ,
|How to cite this URL:|
Maekawa LE, Nassri MG, Ishikawa CK, Martins C, Chung A, Koga-Ito CY. In vitro antimicrobial activity of AH Plus, EndoREZ and Epiphany against microorganisms. Indian J Dent Res [serial online] 2012 [cited 2019 Oct 23];23:469-72. Available from: http://www.ijdr.in/text.asp?2012/23/4/469/104951
Endodontic sealers are essential for the good marginal sealing between gutta-percha and dentine. The features of a good endodontic sealer are to be biocompatible, able to induce repair and mineralization, low viscosity, penetration in the dentinal tubules, low solubility and disintegration. 
Nowadays, many resin-based endodontic sealers have been used in the endodontic practice due to their higher adhesion to the dentinal walls, decreasing the microinfiltration. Sealers such as AH Plus, Endo Rez and Epiphany, showed lower microinfiltration in comparison with zinc oxide and eugenol-based sealer. 
AH Plus showed low solubility and disintegration and good adhesion, besides of the satisfactory biological results in teeth with live pulp. ,, EndoREZ containing biocompatible metacrilate showed the ability to penetrate the dentinal tubules due to its hidrophilic feature.  Moreover, previous clinical studies showed that this material is well tolerated by the periodontal ligament.  Epiphany, a termoplastic synthetic resinous polymer, has been used frequently in the obturation of canals.  This sealer is biocompatible and more resistant to microinfiltration when compared to other obturation systems using gutta-percha. 
Nevertheless, considering that microbial remaining after the biomechanical preparation and intracanal medication are essential factors for the development and persistence of periapical lesions, and therefore for the failing of the endodontic treatment, the antimicrobial activity is an essential feature for sealers for obturation. , The aim of the present study was to evaluate the antimicrobial activity of three endodontic sealers AH Plus, EndoREZ and Epiphany.
| Material and Methods|| |
Three endodontic sealers were evaluated in this study: AH Plus, Epiphany and EndoREZ [Table 1].
The following reference strains were included in this study: Candida albicans, Enterococcus faecalis, Escherichia coli and Staphylococcus aureus [Table 2].
|Table 2: Strains, source, wavelength and optical density adopted for obtaining of standardized suspension (106 cells/ml)|
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Firstly, four isolates were plated in Petri plates containing Mueller Hinton (Difco, Detroit, USA) and incubated at 37°C for 24 h. Then, standardized suspensions (106 cells/ml) of each microorganism in sterile saline solution (NaCl 0,9%) were obtained by spectrophotometry (Micronal S/A - São Paulo, SP, Brasil) [Table 2].
Antimicrobial activity of the materials was evaluated by the method of agar diffusion. A base layer of 10 ml of Mueller Hinton agar was poured in Petri plates (90 x 15 mm). After the solidification of the agar, a second layer or seed layer containing 10 ml of Mueller Hinton agar and 200 μl of the microbial standardized suspensions was poured on the first layer.
After solidification of the second layer, three wells of 6 mm of diameter (one for each material) were obtained by removing a standardized portion of the agar in equidistant points with the aid of sterilized plastic straw. The wells were immediately filled with the sealers to be evaluated. The endodontic sealers AHPlus, EndoREZ and Epiphany were prepared according to the manufacturer's instruction. EndoREZ and Epiphany are provided in individuals tubes with a system that mix the sealer in the moment of the use.
Preparation of AH Plus was performed in equal proportions 1:1 in sterilized glass plate. The sealer was carefully put into the wells with the aid of sterilized spatula. After the distribution of the sealers, Petri plates were incubated at 37°C for 24 h.
Inhibition halos formed around the wells were measured with the aid of a millimetric rule with 0.5 mm of accuracy. All the experiments were repeated five times in different moments.
Data were not compared statistically due to the different diffusibility of the tested substances.  Due to this feature, the inhibition halo does not reflect the efficacy of the antimicrobial agent and the activity of each material was evaluated considering the presence of absence of the inhibition halo. 
| Results|| |
Mean and standard deviation of the inhibition halos (in mm) obtained for each endodontic sealer tested after the period of 24 h are described in [Table 3].
According to [Table 3], Epiphany did not show any antimicrobial activity on the tested microorganisms (without inhibition halo). The AH Plus showed the greatest inhibition halo (14.96 mm) on C. albicans followed by EndoREZ on S. aureus (14.00 mm). EndoREZ also showed greater inhibition halo in comparison to AH Plus on E. faecalis and E. coli. Among all the tested microorganisms, E. faecalis showed higher resistance to the evaluated sealers.
| Discussion|| |
Endodontics sealers are essential to the satisfactory marginal sealing and the criteria of a good sealer are to be biocompatible, dimensionally stable, show good flow, low solubility and disintegration, besides of having antimicrobial activity. 
Antimicrobial activity of sealers might help to eliminate residual microorganisms that have survived the chemo-mechanical instrumentation and thereby improve the success rate of endodontic treatment. 
Many studies have been performed to evaluate the antimicrobial activity of different endodontic sealers and the most frequently used method to evaluate this activity is the diffusion in agar. ,,,,, Nevertheless, this method does not depend only on the toxicity of the material against one microorganism, but it can be influenced by the diffusion and affinity of the substance in the culture media.  A material with easier diffusion consequently will produce greater inhibition halos. 
Regarding the microorganism C. albicans, AH Plus showed the greatest inhibition halo in relation to the other microorganisms tested. EndoREZ showed higher inhibition halos when compared to AH Plus.
A study evaluating 10 endodontic sealers on C. albicans and E. faecalis using the agar diffusion method, showed that AH Plus did not show antimicrobial activity in any of the evaluation periods: 24, 48, 72 h, 5 and 7 days.  Despite of this, another study,  using the same method reported that AH Plus showed antimicrobial activity on C. albicans, S. aureus and E. coli. Besides, Yasuda et al,  verified that AH Plus showed higher antimicrobial activity against all the tested microorganisms (S. aureus, E. faecalis, C. albicans, Streptococcus mutans, Streptococcus saguinis).
The antimicrobial activity showed by AH Plus seems to be correlated to the components epoxy resin and amines that are present in this sealer.  Pizzo et al,  reported that in DCT only fresh AH Plus showed antibacterial activity, whereas 24-hour and 7-day-old samples did not show antibacterial effect against E. faecalis. Similar results were reported by Kayaoglu et al. Estudo de Zhang et al,  also showed that fresh AH Plus had significant antibacterial effect, whereas set samples did not show antimicrobial activity.
Epiphany SE (self-etch) sealer and EndoREZ are dual-cure hydrophilic methacrylate resin-based endodontic sealers.  Several studies verified that EndoREZ sealer did not show antimicrobial activity on E. faecalis, E. coli, S. aureus, C. albicans, Micrococcus luteus, Staphylococcus epidermidis and Pseudomonas aeruginosa. , Nevertheless in this study, the results showed that EndoREZ showed antimicrobial activity on all the studied microorganisms.
Using the modified direct contact test, Zhang et al,  verified EndoREZ demonstrated strong antibacterial effect against E. faecalis throughout the 7-day testing period, and all bacteria were killed during 5-20 minutes of contact with the sealer. EndoREZ was clearly sticky with a moist surface even 7 days after mixing, which indicates that the setting of the sealer was not yet complete at this point. The same authors also reported that incubation of E. faecalis for 1 hour at pH 3 and 3.5 showed that low pH alone does not have an impact on its viability. Slow setting, elution of non reacted monomers, and the lowest pH (below 4) are probably important for the continuing antibacterial effect of EndoREZ.
Another study evaluating the antimicrobial activity of nine endodontic sealers on S. aureus, C. albicans and E. faecalis, by the agar diffusion test verified that EndoREZ and AH Plus showed the lowest antimicrobial activity. 
This lack of agreement among the results can be correlated to variations in the concentrations of both inoculum and tested microorganisms, quantity and curing time of the endodontic sealers tested, incubation period and the evaluation of methods. 
Epiphany sealer did not show antimicrobial activity on the tested microorganisms, considering that no inhibition halo was observed. These results agree with the studies of Pinheiro et al and Bodrumlu and Semiz,  who observed no or low antimicrobial activity of Epiphany on E. faecalis using agar diffusion method.
| Conclusion|| |
According to the methodology, it could be concluded that AH Plus and EndoREZ sealers showed antimicrobial activity against all the tested microorganisms. No antimicrobial activity was observed for Epiphany sealer.
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Department of Restorative Dentistry, School of Dentistry of São José dos Campos, São Paulo State University - UNESP, São José dos Campos, São Paulo
Source of Support: None, Conflict of Interest: None
[Table 1], [Table 2], [Table 3]
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