, Volume 107, Issue 4, pp 513–520 | Cite as

Comparison of the antibacterial activity of calcium silicate- and epoxy resin-based endodontic sealers against Enterococcus faecalis biofilms: a confocal laser-scanning microscopy analysis

  • Sara AlsubaitEmail author
  • Shahad Albader
  • Norah Alajlan
  • Nouf Alkhunaini
  • Abdurahman Niazy
  • Ahmed Almahdy
Original Article


This study assessed the antibacterial activity of BioRoot RCS in comparison with that of the Totalfill BC and AH Plus sealers against Enterococcus faecalis biofilms in dentinal tubules using confocal laser-scanning microscopy. Sixty-six root dentin halves were prepared and sterilized. Three sections were used to ensure sterilization. The remaining were inoculated with E. faecalis. Three specimens were examined to verify the viability of biofilms. The sixty specimens were randomly divided into four groups: AH Plus, BioRoot RCS, Totalfill BC sealer, and no sealer. The specimens were incubated for 1, 7, and 30 days. The specimens were stained and four corners of each disc were scanned. Statistical analysis was performed using two-way ANOVA and Tukey’s post hoc test. Almost half of the bacteria were dead in BioRoot RCS group on day 1 and in Totalfill BC group on day 7. All sealers killed significantly more bacteria than the control after 30 days (P < .05). On day 7, Totalfill BC showed a significantly higher percentage of dead bacteria than BioRoot RCS (P < .05). On day 30, the BioRoot RCS group registered the highest percentage of dead cells (61.75%), which was significantly higher than the percentages of the AH Plus and Totalfill BC groups (P < .05). Calcium silicate-based root canal sealers exerted antimicrobial effects against E. faecalis biofilms. The antibacterial activity of BioRoot RCS was significantly higher than that of the Totalfill BC and AH Plus sealers after 30 days of exposure.


Biofilm Bioceramic Confocal laser-scanning microscopy Dentin Root canal sealer 



This work was approved by the Institutional Review Board of King Saud University in Riyadh, Saudi Arabia (E-17-2687), and the College of Dentistry Research Center (IR0248) of King Saud University in Riyadh, Saudi Arabia. The project was carried out in the Molecular and Cell Biology Laboratory, a core research facility of the King Saud University, College of Dentistry in collaboration with the Prince Naif bin AbdulAziz Health Research Center.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.


  1. 1.
    Kakehashi S, Stanley HR, Fitzgerald RJ. The effects of surgical exposures of dental pulps in germ-free and conventional laboratory rats. Oral Surg Oral Med Oral Pathol. 1965;20:340–9.CrossRefGoogle Scholar
  2. 2.
    Ørstavik D. Materials used for root canal obturation: technical, biological and clinical testing. Endod Topics. 2005;12:25–38.CrossRefGoogle Scholar
  3. 3.
    Kayaoglu G, Erten H, Alacam T, Orstavik D. Short-term antibacterial activity of root canal sealers towards Enterococcus faecalis. Int Endod J. 2005;38:483–8.CrossRefGoogle Scholar
  4. 4.
    Sousa-Neto MD, Silva Coelho FI, Marchesan MA, Alfredo E, Silva-Sousa YT. Ex vivo study of the adhesion of an epoxy-based sealer to human dentine submitted to irradiation with Er:YAG and Nd:YAG lasers. Int Endod J. 2005;38:866–70.CrossRefGoogle Scholar
  5. 5.
    Miletic I, Devcic N, Anic I, Borcic J, Karlovic Z, Osmak M. The cytotoxicity of RoekoSeal and AH plus compared during different setting periods. J Endod. 2005;31:307–9.CrossRefGoogle Scholar
  6. 6.
    Zhou HM, Du TF, Shen Y, Wang ZJ, Zheng YF, Haapasalo M. In vitro cytotoxicity of calcium silicate-containing endodontic sealers. J Endod. 2015;41:56–61.CrossRefGoogle Scholar
  7. 7.
    Viapiana R, Guerreiro-Tanomaru JM, Hungaro-Duarte MA, Tanomaru-Filho M, Camilleri J. Chemical characterization and bioactivity of epoxy resin and Portland cement-based sealers with niobium and zirconium oxide radiopacifiers. Dent Mater. 2014;30:1005–20.CrossRefGoogle Scholar
  8. 8.
    Candeiro GT, Correia FC, Duarte MA, Ribeiro-Siqueira DC, Gavini G. Evaluation of radiopacity, pH, release of calcium ions, and flow of a bioceramic root canal sealer. J Endod. 2012;38:842–5.CrossRefGoogle Scholar
  9. 9.
    Camps J, Jeanneau C, El Ayachi I, Laurent P, About I. Bioactivity of a calcium silicate-based endodontic cement (BioRoot RCS): interactions with human periodontal ligament cells in vitro. J Endod. 2015;41:1469–73.CrossRefGoogle Scholar
  10. 10.
    Eldeniz AU, Shehata M, Högg C, Reichl FX. DNA double-strand breaks caused by new and contemporary endodontic sealers. Int Endod J. 2016;49:1141–51.CrossRefGoogle Scholar
  11. 11.
    Siboni F, Taddei P, Zamparini F, Prati C, Gandolfi MG. Properties of BioRoot RCS, a tricalcium silicate endodontic sealer modified with povidone and polycarboxylate. Int Endod J. 2017;50(Suppl 2):e120-e36.Google Scholar
  12. 12.
    Viapiana R, Moinzadeh AT, Camilleri L, Wesselink PR, Tanomaru Filho M, Camilleri J. Porosity and sealing ability of root fillings with gutta-percha and BioRoot RCS or AH Plus sealers. Evaluation by three ex vivo methods. Int Endod J. 2016;49:774–82.CrossRefGoogle Scholar
  13. 13.
    Uzunoglu-Özyürek E, Erdoğan Ö, Aktemur Türker S. Effect of calcium hydroxide dressing on the dentinal tubule penetration of 2 different root canal sealers: a confocal laser scanning microscopic study. J Endod. 2018;44:1018–23.CrossRefGoogle Scholar
  14. 14.
    Abdulkader A, Duguid R, Saunders EM. The antimicrobial activity of endodontic sealers to anaerobic bacteria. Int Endod J. 1996;29:280–3.CrossRefGoogle Scholar
  15. 15.
    Siqueira JF Jr, Favieri A, Gahyva SM, Moraes SR, Lima KC, Lopes HP. Antimicrobial activity and flow rate of newer and established root canal sealers. J Endod. 2000;26:274–7.CrossRefGoogle Scholar
  16. 16.
    Candeiro GT, Moura-Netto C, D’Almeida-Couto RS, Azambuja-Junior N, Marques MM, Cai S, Gavini G. Cytotoxicity, genotoxicity and antibacterial effectiveness of a bioceramic endodontic sealer. Int Endod J. 2016;49:858–64.CrossRefGoogle Scholar
  17. 17.
    Weiss EI, Shalhav M, Fuss Z. Assessment of antibacterial activity of endodontic sealers by a direct contact test. Endod Dent Traumatol. 1996;12:179–84.CrossRefGoogle Scholar
  18. 18.
    Wang Z, Shen Y, Haapasalo M. Dentin extends the antibacterial effect of endodontic sealers against Enterococcus faecalis biofilms. J Endod. 2014;40:505–8.CrossRefGoogle Scholar
  19. 19.
    Haapasalo M, Qian W, Portenier I, Waltimo T. Effects of dentin on the antimicrobial properties of endodontic medicaments. J Endod. 2007;33:917–25.CrossRefGoogle Scholar
  20. 20.
    Morgental RD, Singh A, Sappal H, Kopper PM, Vier-Pelisser FV, Peters OA. Dentin inhibits the antibacterial effect of new and conventional endodontic irrigants. J Endod. 2013;39:406–10.CrossRefGoogle Scholar
  21. 21.
    Ma J, Wang Z, Shen Y, Haapasalo M. A new noninvasive model to study the effectiveness of dentin disinfection by using confocal laser scanning microscopy. J Endod. 2011;37:1380–5.CrossRefGoogle Scholar
  22. 22.
    Stuart CH, Schwartz SA, Beeson TJ, Owatz CB. Enterococcus faecalis: its role in root canal treatment failure and current concepts in retreatment. J Endod. 2006;32:93–8.CrossRefGoogle Scholar
  23. 23.
    Sundqvist G, Figdor D, Persson S, Sjogren U. Microbiologic analysis of teeth with failed endodontic treatment and the outcome of conservative re-treatment. Oral Surg Oral Med Oral Pathol Oral Radiol Endod. 1998;85:86–93.CrossRefGoogle Scholar
  24. 24.
    George S, Kishen A, Song KP. The role of environmental changes on monospecies biofilm formation on root canal wall by Enterococcus faecalis. J Endod. 2005;31:867–72.CrossRefGoogle Scholar
  25. 25.
    Schäfer E, Bössmann K. Antimicrobial efficacy of chlorhexidine and two calcium hydroxide formulations against Enterococcus faecalis. J Endod. 2005;31:53–6.CrossRefGoogle Scholar
  26. 26.
    Lima RK, Guerreiro-Tanomaru JM, Faria-Junior NB, Tanomaru-Filho M. Effectiveness of calcium hydroxide-based intracanal medicaments against Enterococcus faecalis. Int Endod J. 2012;45:311–6.CrossRefGoogle Scholar
  27. 27.
    Topcuoglu HS, Tuncay O, Karatas E, Arslan H, Yeter K. In vitro fracture resistance of roots obturated with epoxy resin-based, mineral trioxide aggregate-based, and bioceramic root canal sealers. J Endod. 2013;39:1630–3.CrossRefGoogle Scholar
  28. 28.
    White JM, Goodis HE, Marshall SJ, Marshall GW. Sterilization of teeth by gamma radiation. J Dent Res. 1994;73:1560–7.CrossRefGoogle Scholar
  29. 29.
    Ricucci D, Siqueira JF Jr. Biofilms and apical periodontitis: study of prevalence and association with clinical and histopathologic findings. J Endod. 2010;36:1277–88.CrossRefGoogle Scholar
  30. 30.
    Upadya MH, Kishen A. Influence of bacterial growth modes on the susceptibility to light-activated disinfection. Int Endod J. 2010;43:978–87.CrossRefGoogle Scholar
  31. 31.
    Wang Z, Shen Y, Haapasalo M. Effectiveness of endodontic disinfecting solutions against young and old Enterococcus faecalis biofilms in dentin canals. J Endod. 2012;38:1376–9.CrossRefGoogle Scholar
  32. 32.
    Watson TF, Cook RJ, Festy F, Pilecki P, Sauro S. Optical imaging techniques for dental biomaterials interfaces. In: Curtis RV, Watson TF, editors. Dental biomaterials: imaging, testing and modelling. Cambridge: Woodhead; 2008.CrossRefGoogle Scholar
  33. 33.
    Pizzo G, Giammanco GM, Cumbo E, Nicolosi G, Gallina G. In vitro antibacterial activity of endodontic sealers. J Dent. 2006;34:35–40.CrossRefGoogle Scholar
  34. 34.
    Zhang H, Shen Y, Ruse ND, Haapasalo M. Antibacterial activity of endodontic sealers by modified direct contact test against Enterococcus faecalis. J Endod. 2009;35:1051–5.CrossRefGoogle Scholar
  35. 35.
    Schweikl H, Schmalz G. The induction of micronuclei in V79 cells by the root canal filling material AH plus. Biomaterials. 2000;21:939–44.CrossRefGoogle Scholar
  36. 36.
    Leonardo MR, Bezerra da Silva LA, Filho MT, Santana da Silva R. Release of formaldehyde by 4 endodontic sealers. Oral Surg Oral Med Oral Pathol Oral Radiol Endod. 1999;88:221–5.CrossRefGoogle Scholar
  37. 37.
    Al-Haddad A, Che Ab Aziz ZA. Bioceramic-based root canal sealers: a review. Int J Biomater. 2016;2016:9753210.CrossRefGoogle Scholar
  38. 38.
    Trope M, Bunes A, Debelian G. Root filling materials and techniques: bioceramics a new hope? Endod Topics. 2015;32:86–96.CrossRefGoogle Scholar
  39. 39.
    Zhou HM, Shen Y, Zheng W, Li L, Zheng YF, Haapasalo M. Physical properties of 5 root canal sealers. J Endod. 2013;39:1281–6.CrossRefGoogle Scholar
  40. 40.
    Borges RP, Sousa-Neto MD, Versiani MA, Rached-Junior FA, De-Deus G, Miranda CE, Pecora JD. Changes in the surface of four calcium silicate-containing endodontic materials and an epoxy resin-based sealer after a solubility test. Int Endod J. 2012;45:419–28.CrossRefGoogle Scholar
  41. 41.
    Alsubait SA, Al Ajlan R, Mitwalli H, Aburaisi N, Mahmood A, Muthurangan M, Almadhri R, Alfayez M, Anil S. Cytotoxicity of different concentrations of three root canal sealers on human mesenchymal stem cells. Biomolecules. 2018;1:8:E68.CrossRefGoogle Scholar
  42. 42.
    Khalil I, Naaman A, Camilleri J. Properties of tricalcium silicate sealers. J Endod. 2016;42:1529–35.CrossRefGoogle Scholar
  43. 43.
    Arias-Moliz MT, Camilleri J. The effect of the final irrigant on the antimicrobial activity of root canal sealers. J Dent. 2016;52:30–6.CrossRefGoogle Scholar
  44. 44.
    Du T, Wang Z, Shen Y, Ma J, Cao Y, Haapasalo M. Effect of long-term exposure to endodontic disinfecting solutions on young and old Enterococcus faecalis biofilms in dentin canals. J Endod. 2014;40:509–14.CrossRefGoogle Scholar
  45. 45.
    Chivatxaranukul P, Dashper SG, Messer HH. Dentinal tubule invasion and adherence by Enterococcus faecalis. Int Endod J. 2008;41:873–82.CrossRefGoogle Scholar

Copyright information

© The Society of The Nippon Dental University 2019

Authors and Affiliations

  • Sara Alsubait
    • 1
    Email author
  • Shahad Albader
    • 2
  • Norah Alajlan
    • 2
  • Nouf Alkhunaini
    • 2
  • Abdurahman Niazy
    • 3
  • Ahmed Almahdy
    • 4
  1. 1.Department of Restorative Dental Science, College of DentistryKing Saud UniversityRiyadhSaudi Arabia
  2. 2.General PractitionerRiyadhSaudi Arabia
  3. 3.Department of Oral Medicine and Diagnostic Sciences, College of DentistryKing Saud UniversityRiyadhSaudi Arabia
  4. 4.Department of Pediatric Dentistry and Orthodontics, College of DentistryKing Saud UniversityRiyadhSaudi Arabia

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