In Vitro Antimicrobial Effects of Endoseal MTA and AH Plus Sealers on Enterococcus Faecalis and Candida Albicans Mature Biofilms
Abstract
Objectives: This study compared the antimicrobial effects of Endoseal MTA and AH Plus sealers on Enterococcus faecalis (E. faecalis) and Candida albicans (C. albicans) mature biofilms on dentin blocks.
Materials and Methods: In this in vitro study, the root canals of 207 dentin blocks sectioned from the roots of extracted teeth were inoculated with E. faecalis and C. albicans, and incubated for 3 weeks. The specimens were evaluated in 12 groups (n=17) for random root canal filling with AH Plus, Endoseal MTA, or no sealer, and assessment after 24 hours and 2 weeks. Dentin chips were cultured on blood agar, and the colonies were counted. One random specimen from each group was assessed under a scanning electron microscope (SEM). Data were analyzed using the Kruskal-Wallis and Mann-Whitney tests with Bonferroni adjustment at 0.05 level of significance.
Results: At 24 hours, Endoseal MTA significantly decreased the E. faecalis count compared with the positive control (P<0.008). At 2 weeks, the colony count of both microorganisms was significantly lower in the AH Plus group compared with the control group (P<0.008). The C. albicans colony count in the Endoseal MTA group was significantly lower than that in the control group at 2 weeks (P<0.008). AH Plus caused significantly greater reduction in C. albicans colony count at 2 weeks compared with Endoseal MTA (P<0.008).
Conclusion: AH Plus showed a higher antimicrobial activity over time against both E. faecalis and C. albicans. Endoseal MTA only showed short-term antibacterial effect.
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2. Upadya MH, Kishen A. Influence of bacterial growth modes on the susceptibility to light‐activated disinfection. Int Endod J. 2010 Nov;43(11):978-87.
3. Hala F, Mohammed M, Thanaa I. Antibacterial effect of two types of nano particles incorporated in zinc oxide based sealers on Enterococcus faecalis (in vitro study). Alex Dent J. 2016 Aug;41(2):169-75.
4. Waltimo TM, Siren EK, Torkko HL, Olsen I, Haapasalo MP. Fungi in therapy‐resistant apical periodontitis. Int Endod J. 1997 Mar;30(2):96-101.
5. Waltimo TM, Sirén EK, Orstavik D, Haapasalo MP. Susceptibility of oral Candida species to calcium hydroxide in vitro. Int Endod J. 1999 Mar;32(2):94-8.
6. Şen BH, Safavi KE, Spångberg LS. Growth patterns of Candida albicans in relation to radicular dentin. Oral Surg, Oral Med, Oral Pathol, Oral Radiol, Endod. 1997 Jul;84(1):68-73.
7. Waltimo TM, Haapasalo M, Zehnder M, Meyer J. Clinical aspects related to endodontic yeast infections. Endod Topics. 2004 Nov;9(1):66-78.
8. Anderl JN, Franklin MJ, Stewart PS. Role of antibiotic penetration limitation in Klebsiella pneumoniae biofilm resistance to ampicillin and ciprofloxacin. Antimicrob Agents Chemother. 2000 Jul;44(7):1818-24.
9. Zhang H, Shen Y, Ruse ND, Haapasalo M. Antibacterial activity of endodontic sealers by modified direct contact test against Enterococcus faecalis. J Endod. 2009 Jul;35(7):1051-5.
10. Pasqualini D, Cuffini AM, Scotti N, Mandras N, Scalas D, Pera F, Berutti E. Comparative evaluation of the antimicrobial efficacy of a 5% sodium hypochlorite subsonic-activated solution. J Endod. 2010 Aug 1;36(8):1358-60.
11. Wang Z. Bioceramic materials in endodontics. Endod Topics. 2015 May;32(1):3-0.
12. Hajipour MJ, Fromm KM, Ashkarran AA, de Aberasturi DJ, de Larramendi IR, Rojo T, Serpooshan V, Parak WJ, Mahmoudi M. Antibacterial properties of nanoparticles. Trends Biotechnol. 2012 Oct;30(10):499-511.
13. Lim ES, Park YB, Kwon YS, Shon WJ, Lee KW, Min KS. Physical properties and biocompatibility of an injectable calcium-silicate-based root canal sealer: in vitro and in vivo study. BMC Oral Health. 2015 Dec;15:1-7.
14. Donlan RM, Costerton JW. Biofilms: survival mechanisms of clinically relevant microorganisms. Clin Microbiol Rev. 2002 Apr;15(2):167-93.
15. Haapasalo M, Ørstavik D. In vitro infection and of dentinal tubules. J Dent Res. 1987 Aug;66(8):1375-9.
16. Bukhari S, Karabucak B. The antimicrobial effect of bioceramic sealer on an 8-week matured Enterococcus faecalis biofilm attached to root canal dentinal surface. J Endod. 2019 Aug;45(8):1047-52.
17. Shaik J, Garlapati R, Nagesh B, Sujana V, Jayaprakash T, Naidu S. Comparative evaluation of antimicrobial efficacy of triple antibiotic paste and calcium hydroxide using chitosan as carrier against Candida albicans and Enterococcus faecalis: an: in vitro: study. J Conserv Dent Endod. 2014 Jul;17(4):335-9.
18. Vanapatla A, Vemisetty H, Punna R, Veeramachineni C, Venkata RP, Muppala JN, Dandolu R. Comparative evaluation of antimicrobial effect of three endodontic sealers with and without antibiotics–An In-vitro Study. J Clin Diagnostic Res. 2016 Apr;10(4):ZC69.
19. Brown JH, Brenn L. A method for the differential staining of Gram-positive and Gram-negative bacteria in tissue sections. Bull. Johns Hopkins Hosp. 1931 Feb;48:69-73.
20. Gürel M, Demiryürek EÖ, Özyürek T, Gülhan T. Antimicrobial activities of different bioceramic root canal sealers on various bacterial species. Int J Appl Dent Sci. 2016;2(3):19-22.
21. de Miranda 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 Jun;38(6):842-5.
22. Bodrumlu E, Semiz M. Antibacterial activity of a new endodontic sealer against Enterococcus faecalis. J Can Dent Assoc. 2006 Sep;72(7):637.
23. Alsubait S, Albader S, Alajlan N, Alkhunaini N, Niazy A, Almahdy A. Comparison of the antibacterial activity of calcium silicate-and epoxy resin-based endodontic sealers against Enterococcus faecalis biofilms: a confocal laser-scanning microscopy analysis. Odontology. 2019 Oct;107:513-20.
24. Özcan E, Yula E, Arslanoğlu Z, İnci M. Antifungal activity of several root canal sealers against Candida albicans. Acta Odontol Scand. 2013 Nov;71(6):1481-5.
25. Pizzo G, Giammanco GM, Cumbo E, Nicolosi G, Gallina G. In vitro antibacterial activity of endodontic sealers. J Dent. 2006 Jan;34(1):35-40.
26. Kayaoglu GÜ, Erten H, Alaçam TA, Ørstavik D. Short‐term antibacterial activity of root canal sealers towards Enterococcus faecalis. Int Endod J. 2005 Jul;38(7):483-8.
27. Candeiro GD, Moura‐Netto CD, D'Almeida‐Couto RS, Azambuja‐Júnior N, Marques MM, Cai S, Gavini G. Cytotoxicity, genotoxicity and antibacterial effectiveness of a bioceramic endodontic sealer. Int Endod J. 2016 Sep;49(9):858-64.
28. Saleh IM, Ruyter IE, Haapasalo M, Ørstavik D. Survival of Enterococcus faecalis in infected dentinal tubules after root canal filling with different root canal sealers in vitro. Int Endod J. 2004 Mar;37(3):193-8.
29. Gomes BP, Pinheiro ET, Gadê‐Neto CR, Sousa EL, Ferraz CC, Zaia AA, Teixeira FB, Souza‐Filho FJ. Microbiological examination of infected dental root canals. Oral Microbiol Immunol. 2004 Apr;19(2):71-6.
30. Cohen BI, Pagnillo MK, Musikant BL, Deutsch AS. Formaldehyde evaluation from endodontic materials. Oral Health. 1998 Dec;88(12):37-9.
31. Mickel AK, Nguyen TH, Chogle S. Antimicrobial activity of endodontic sealers on Enterococcus faecalis. J Endod. 2003 Apr;29(4):257-8.
32. Slutzky-Goldberg I, Slutzky H, Solomonov M, Moshonov J, Weiss EI, Matalon S. Antibacterial properties of four endodontic sealers. J Endod. 2008 Jun;34(6):735-8.
33. Loushine BA, Bryan TE, Looney SW, Gillen BM, Loushine RJ, Weller RN, Pashley DH, Tay FR. Setting properties and cytotoxicity evaluation of a premixed bioceramic root canal sealer. J Endod. 2011 May 1;37(5):673-7.
34. Mak ST, Leong XF, Tew IM, Kumolosasi E, Wong L. In vitro evaluation of the antibacterial activity of EndoSeal MTA, iRoot SP, and AH Plus against planktonic bacteria. Materials. 2022 Mar 8;15(6):2012.
35. Shin JH, Lee DY, Lee SH. Comparison of antimicrobial activity of traditional and new developed root sealers against pathogens related root canal. J Dent Sci. 2018 Mar;13(1):54-9.
36. Estrela C, Sousa-Neto MD, Alves DR, Alencar AH, Santos TO, Pécora JD. A preliminary study of the antibacterial potential of cetylpyridinium chloride in root canals infected by E. faecalis. Braz Dent J. 2012;23:645-53.
37. Kaya S, Yiğit-Özer S, Adigüzel Ö. Evaluation of radicular dentin erosion and smear layer removal capacity of Self-Adjusting File using different concentrations of sodium hypochlorite as an initial irrigant. Oral Surg, Oral Med, Oral Pathol, Oral Radiol, Endod. 2011 Oct;112(4):524-30.
38. Wang Z, Shen Y, Haapasalo M. Effectiveness of endodontic disinfecting solutions against young and old Enterococcus faecalis biofilms in dentin canals. J Endod. 2012 Oct;38(10):1376-9.
39. Bergenholtz G. Micro-organisms from necrotic pulp of traumatized teeth. Odontol Revy. 1974;25(4):347-58.
40. Burmølle M, Webb JS, Rao D, Hansen LH, Sørensen SJ, Kjelleberg S. Enhanced biofilm formation and increased resistance to antimicrobial agents and bacterial invasion are caused by synergistic interactions in multispecies biofilms. Appl Environ Microbiol. 2006 Jun;72(6):3916-23.
2. Upadya MH, Kishen A. Influence of bacterial growth modes on the susceptibility to light‐activated disinfection. Int Endod J. 2010 Nov;43(11):978-87.
3. Hala F, Mohammed M, Thanaa I. Antibacterial effect of two types of nano particles incorporated in zinc oxide based sealers on Enterococcus faecalis (in vitro study). Alex Dent J. 2016 Aug;41(2):169-75.
4. Waltimo TM, Siren EK, Torkko HL, Olsen I, Haapasalo MP. Fungi in therapy‐resistant apical periodontitis. Int Endod J. 1997 Mar;30(2):96-101.
5. Waltimo TM, Sirén EK, Orstavik D, Haapasalo MP. Susceptibility of oral Candida species to calcium hydroxide in vitro. Int Endod J. 1999 Mar;32(2):94-8.
6. Şen BH, Safavi KE, Spångberg LS. Growth patterns of Candida albicans in relation to radicular dentin. Oral Surg, Oral Med, Oral Pathol, Oral Radiol, Endod. 1997 Jul;84(1):68-73.
7. Waltimo TM, Haapasalo M, Zehnder M, Meyer J. Clinical aspects related to endodontic yeast infections. Endod Topics. 2004 Nov;9(1):66-78.
8. Anderl JN, Franklin MJ, Stewart PS. Role of antibiotic penetration limitation in Klebsiella pneumoniae biofilm resistance to ampicillin and ciprofloxacin. Antimicrob Agents Chemother. 2000 Jul;44(7):1818-24.
9. Zhang H, Shen Y, Ruse ND, Haapasalo M. Antibacterial activity of endodontic sealers by modified direct contact test against Enterococcus faecalis. J Endod. 2009 Jul;35(7):1051-5.
10. Pasqualini D, Cuffini AM, Scotti N, Mandras N, Scalas D, Pera F, Berutti E. Comparative evaluation of the antimicrobial efficacy of a 5% sodium hypochlorite subsonic-activated solution. J Endod. 2010 Aug 1;36(8):1358-60.
11. Wang Z. Bioceramic materials in endodontics. Endod Topics. 2015 May;32(1):3-0.
12. Hajipour MJ, Fromm KM, Ashkarran AA, de Aberasturi DJ, de Larramendi IR, Rojo T, Serpooshan V, Parak WJ, Mahmoudi M. Antibacterial properties of nanoparticles. Trends Biotechnol. 2012 Oct;30(10):499-511.
13. Lim ES, Park YB, Kwon YS, Shon WJ, Lee KW, Min KS. Physical properties and biocompatibility of an injectable calcium-silicate-based root canal sealer: in vitro and in vivo study. BMC Oral Health. 2015 Dec;15:1-7.
14. Donlan RM, Costerton JW. Biofilms: survival mechanisms of clinically relevant microorganisms. Clin Microbiol Rev. 2002 Apr;15(2):167-93.
15. Haapasalo M, Ørstavik D. In vitro infection and of dentinal tubules. J Dent Res. 1987 Aug;66(8):1375-9.
16. Bukhari S, Karabucak B. The antimicrobial effect of bioceramic sealer on an 8-week matured Enterococcus faecalis biofilm attached to root canal dentinal surface. J Endod. 2019 Aug;45(8):1047-52.
17. Shaik J, Garlapati R, Nagesh B, Sujana V, Jayaprakash T, Naidu S. Comparative evaluation of antimicrobial efficacy of triple antibiotic paste and calcium hydroxide using chitosan as carrier against Candida albicans and Enterococcus faecalis: an: in vitro: study. J Conserv Dent Endod. 2014 Jul;17(4):335-9.
18. Vanapatla A, Vemisetty H, Punna R, Veeramachineni C, Venkata RP, Muppala JN, Dandolu R. Comparative evaluation of antimicrobial effect of three endodontic sealers with and without antibiotics–An In-vitro Study. J Clin Diagnostic Res. 2016 Apr;10(4):ZC69.
19. Brown JH, Brenn L. A method for the differential staining of Gram-positive and Gram-negative bacteria in tissue sections. Bull. Johns Hopkins Hosp. 1931 Feb;48:69-73.
20. Gürel M, Demiryürek EÖ, Özyürek T, Gülhan T. Antimicrobial activities of different bioceramic root canal sealers on various bacterial species. Int J Appl Dent Sci. 2016;2(3):19-22.
21. de Miranda 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 Jun;38(6):842-5.
22. Bodrumlu E, Semiz M. Antibacterial activity of a new endodontic sealer against Enterococcus faecalis. J Can Dent Assoc. 2006 Sep;72(7):637.
23. Alsubait S, Albader S, Alajlan N, Alkhunaini N, Niazy A, Almahdy A. Comparison of the antibacterial activity of calcium silicate-and epoxy resin-based endodontic sealers against Enterococcus faecalis biofilms: a confocal laser-scanning microscopy analysis. Odontology. 2019 Oct;107:513-20.
24. Özcan E, Yula E, Arslanoğlu Z, İnci M. Antifungal activity of several root canal sealers against Candida albicans. Acta Odontol Scand. 2013 Nov;71(6):1481-5.
25. Pizzo G, Giammanco GM, Cumbo E, Nicolosi G, Gallina G. In vitro antibacterial activity of endodontic sealers. J Dent. 2006 Jan;34(1):35-40.
26. Kayaoglu GÜ, Erten H, Alaçam TA, Ørstavik D. Short‐term antibacterial activity of root canal sealers towards Enterococcus faecalis. Int Endod J. 2005 Jul;38(7):483-8.
27. Candeiro GD, Moura‐Netto CD, D'Almeida‐Couto RS, Azambuja‐Júnior N, Marques MM, Cai S, Gavini G. Cytotoxicity, genotoxicity and antibacterial effectiveness of a bioceramic endodontic sealer. Int Endod J. 2016 Sep;49(9):858-64.
28. Saleh IM, Ruyter IE, Haapasalo M, Ørstavik D. Survival of Enterococcus faecalis in infected dentinal tubules after root canal filling with different root canal sealers in vitro. Int Endod J. 2004 Mar;37(3):193-8.
29. Gomes BP, Pinheiro ET, Gadê‐Neto CR, Sousa EL, Ferraz CC, Zaia AA, Teixeira FB, Souza‐Filho FJ. Microbiological examination of infected dental root canals. Oral Microbiol Immunol. 2004 Apr;19(2):71-6.
30. Cohen BI, Pagnillo MK, Musikant BL, Deutsch AS. Formaldehyde evaluation from endodontic materials. Oral Health. 1998 Dec;88(12):37-9.
31. Mickel AK, Nguyen TH, Chogle S. Antimicrobial activity of endodontic sealers on Enterococcus faecalis. J Endod. 2003 Apr;29(4):257-8.
32. Slutzky-Goldberg I, Slutzky H, Solomonov M, Moshonov J, Weiss EI, Matalon S. Antibacterial properties of four endodontic sealers. J Endod. 2008 Jun;34(6):735-8.
33. Loushine BA, Bryan TE, Looney SW, Gillen BM, Loushine RJ, Weller RN, Pashley DH, Tay FR. Setting properties and cytotoxicity evaluation of a premixed bioceramic root canal sealer. J Endod. 2011 May 1;37(5):673-7.
34. Mak ST, Leong XF, Tew IM, Kumolosasi E, Wong L. In vitro evaluation of the antibacterial activity of EndoSeal MTA, iRoot SP, and AH Plus against planktonic bacteria. Materials. 2022 Mar 8;15(6):2012.
35. Shin JH, Lee DY, Lee SH. Comparison of antimicrobial activity of traditional and new developed root sealers against pathogens related root canal. J Dent Sci. 2018 Mar;13(1):54-9.
36. Estrela C, Sousa-Neto MD, Alves DR, Alencar AH, Santos TO, Pécora JD. A preliminary study of the antibacterial potential of cetylpyridinium chloride in root canals infected by E. faecalis. Braz Dent J. 2012;23:645-53.
37. Kaya S, Yiğit-Özer S, Adigüzel Ö. Evaluation of radicular dentin erosion and smear layer removal capacity of Self-Adjusting File using different concentrations of sodium hypochlorite as an initial irrigant. Oral Surg, Oral Med, Oral Pathol, Oral Radiol, Endod. 2011 Oct;112(4):524-30.
38. Wang Z, Shen Y, Haapasalo M. Effectiveness of endodontic disinfecting solutions against young and old Enterococcus faecalis biofilms in dentin canals. J Endod. 2012 Oct;38(10):1376-9.
39. Bergenholtz G. Micro-organisms from necrotic pulp of traumatized teeth. Odontol Revy. 1974;25(4):347-58.
40. Burmølle M, Webb JS, Rao D, Hansen LH, Sørensen SJ, Kjelleberg S. Enhanced biofilm formation and increased resistance to antimicrobial agents and bacterial invasion are caused by synergistic interactions in multispecies biofilms. Appl Environ Microbiol. 2006 Jun;72(6):3916-23.
| Issue | Vol 22 (Continuously Published Article-Based) | |
| Section | Original Article | |
| DOI | https://doi.org/10.18502/fid.v22i19.18760 | |
| Keywords | ||
| Root Canal Therapy Enterococcus Faecalis Candida Albicans Anti-Infective Agents | ||
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This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License. |
How to Cite
1.
Khoshbin E, Nikzad F, Arabestani MR, Farhdian M, Karkehabadi H. In Vitro Antimicrobial Effects of Endoseal MTA and AH Plus Sealers on Enterococcus Faecalis and Candida Albicans Mature Biofilms. Front Dent. 2025;22.
