Skip to main content
SpringerLink
Log in

Pathogenic potential of Enterococcus faecalis strains isolated from root canals after unsuccessful endodontic treatment

  • Original Article
  • Published:
Clinical Oral Investigations Aims and scope Submit manuscript

ABSTRACT

Objectives

To evaluate strains of Enterococcus faecalis isolated from endodontic failures cases for (a) presence of virulence genes, namely, gelatinase production (gelE), surface protein (esp), collagen-binding adhesin (ace), cytolysin activator (cylA), E. faecalis antigen A (efaA) and aggregation substance (asa), all by using PCR; (b) biofilm formation capacity; and (c) activity of gelatinase and β-lactamase.

Materials and methods

Twenty-five strains of E. faecalis were tested. The DNA extracted from these strains was used for identification of virulence genes by PCR and 1% agarose gel. Biofilm formation was performed on polystyrene microplates by using the violet crystal staining method. For assessment of the gelatinase activity, inoculum of pure cultures was deposited in tubes containing gelatin and a nutrient broth, whereas nitrocefin disks were used to assess the β-lactamase action.

Results

The virulence genes efaA and cylA were detected in 100% of the strains, whereas gelE was present in 84%, ace in 68%, esp in 56% and asa in 48%. Four strains had no biofilm formation, 17 had poor formation and four had moderate formation. Gelatinase production was observed in three strains and β-lactamase resistance in five strains of E. faecalis.

Topic

Diverse patterns of virulence gene detection were observed among the E. faecalis strains, with predominance of those capable of forming biofilm. A few strains have been found to hydrolyze gelatin proteins, whereas β-lactamase resistance was detected in different isolates.

Clinical relevance

To understand the influence of virulence factors in E. faecalis on the host heath status.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2

Similar content being viewed by others

References

  1. Gomes BP, Pinheiro ET, Jacinto RC, Zaia AA, Ferraz CC, Souza-Filho FJ (2008) Microbial analysis of canals of root-filled teeth with periapical lesions using polymerase chain reaction. J Endod 34:537–540

    Article  PubMed  Google Scholar 

  2. Francisco PA, Delboni MG, Lima AR, Xiao Y, Siqueira WL, Gomes BPFA (2019) Proteomic profile of root canal contents in teeth with post-treatment endodontic disease. Int Endod J 52:451–460

    Article  PubMed  Google Scholar 

  3. Barbosa-Ribeiro M, De-Jesus-Soares A, Zaia AA, Ferraz CC, Almeida JF, Gomes BP (2016) Quantification of lipoteichoic acid contents and cultivable bacteria at the different phases of the endodontic retreatment. J Endod 42:552–556

    Article  PubMed  Google Scholar 

  4. Sundqvist G, Figdor D, Persson S, Sjogren U (1998) Microbiologic analysis of teeth with failed endodontic treatment and the outcome of conservative re-treatment. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 85:86–93

    Article  PubMed  Google Scholar 

  5. Nair PN (2004) Pathogenesis of apical periodontitis and the causes of endodontic failures. Crit Rev Oral Biol Med 15:348–381

    Article  PubMed  Google Scholar 

  6. Rôças IN, Neves MA, Provenzano JC, Siqueira JF (2014) Susceptibility of as-yet-uncultivated and difficult-to-culture bacteria to chemomechanical procedures. J Endod 40:33–37

    Article  PubMed  Google Scholar 

  7. Tzanetakis GN, Azcarate-Peril MA, Zachaki S, Panopoulos P, Kontakiotis EG, Madianos PN, Davaris K (2015) Comparison of bacterial community composition of primary and persistent endodontic infections using pyrosequencing. J Endod 41:1226–1233

    Article  PubMed  PubMed Central  Google Scholar 

  8. Barbosa-Ribeiro M, De-Jesus-Soares A, Zaia AA, Ferraz CC, Almeida JF, Gomes BP (2016) Antimicrobial susceptibility and characterization of virulence genes of Enterococcus faecalis isolates from teeth with failure of the endodontic treatment. J Endod 42:1022–1028

    Article  PubMed  Google Scholar 

  9. Delboni MG, Gomes BP, Francisco PA, Teixeira FB, Drake D (2017) Diversity of Enterococcus faecalis genotypes from multiple oral sites associated with endodontic failure using repetitive sequence-based polymerase chain reaction and arbitrarily primed polymerase chain reaction. J Endod 43:377–382

    Article  PubMed  Google Scholar 

  10. Ozbek SM, Ozbek A, Erdogan AS (2009) Detection of Treponema denticola in symptomatic apical periodontitis and in symptomatic apical abscesses by real-time PCR. Eur J Dent 3:107–113

    Article  PubMed  PubMed Central  Google Scholar 

  11. Guerreiro-Tanomaru JM, de Faria-Junior NB, Duarte MA, Ordinola-Zapata R, Graeff MS (2013 Mar) Tanomaru-Filho M (2013) Comparative analysis of Enterococcus faecalis biofilm formation on different substrates. J Endod 39(3):346–350

    Article  PubMed  Google Scholar 

  12. Kayaoglu G, Orstavik D (2004) Virulence factors of Enterococcus faecalis: relationship to endodontic disease. Crit Rev Oral Biol Med 15:308–320

    Article  PubMed  Google Scholar 

  13. Siqueira JF, Rôças IN (2007) Bacterial pathogenesis and mediators in apical periodontitis. Braz Dent J 18:267–280

    Article  PubMed  Google Scholar 

  14. Medeiros AW, Pereira RI, Oliveira DV, Martins PD, d'Azevedo PA, Van der Sand S, Frazzon J, Frazzon AP (2014) Molecular detection of virulence factors among food and clinical Enterococcus faecalis strains in South Brazil. Braz J Microbiol 45:327–332

    Article  PubMed  PubMed Central  Google Scholar 

  15. Anderson AC, Jonas D, Huber I, Karygianni L, Wolber J, Hellwig E, Arweiler N, Vach K, Wittmer A, Al-Ahmad A (2015) Enterococcus faecalis from food, clinical specimens, and oral sites: prevalence of virulence factors in association with biofilm formation. Front Microbiol 6:1534

    PubMed  Google Scholar 

  16. Usha L (2010) Biofilm in endodontics: new understanding to an old problem. Int J Contem Dent 1:44–51

    Google Scholar 

  17. Portenier I, Waltimo TMT, Haapasalo M (2003) Enterococcus faecalis: the root canal survivor and ‘star’ in post - treatment disease. Endod Top 6:135–159

    Article  Google Scholar 

  18. Ricucci D, Siqueira JF Jr (2010) Biofilms and apical periodontitis: study of prevalence and association with clinical and histopathologic findings. J Endod 36:1277–1288

    Article  PubMed  Google Scholar 

  19. Duggan JM, Sedgley CM (2007) Biofilm formation of oral and endodontic Enterococcus faecalis. J Endod 33:815–818

    Article  PubMed  Google Scholar 

  20. Kishen A, Haapasalo M (2010) Biofilm models and methods of biofilm assessment. Endod Top 22:58–78

    Article  Google Scholar 

  21. Schaechter M, Engleberg NC, Medoff G (1999) Mechanisms of microbial disease, 3rd edn. Lippincott Williams & Wilkins, Philadelphia

    Google Scholar 

  22. Thurlow LR, Thomas VC, Narayanan S, Olson S, Fleming SD, Hancock LE (2010) Gelatinase contributes to the pathogenesis of endocarditis caused by Enterococcus faecalis. Infect Immun 78:4936–4943

    Article  PubMed  PubMed Central  Google Scholar 

  23. Appelbaum PC, Spangler SK, Jacobs MR (1990) Beta-lactamase production and susceptibilities to amoxicillin, amoxicillin-clavulanate, ticarcillin, ticarcillin-clavulanate, cefoxitin, imipenem, and metronidazole of 320 non-Bacteroides fragilis Bacteroides isolates and 129 fusobacteria from 28 U.S. centers. Antimicrob Agents Chemother 34:1546–1550

    Article  PubMed  PubMed Central  Google Scholar 

  24. Morrison D, Woodford N, Cookson B (1997) Enterococci as emerging pathogens of humans. Soc Appl Bacteriol Symp Ser 26:89–99

    Article  Google Scholar 

  25. Pinheiro ET, Gomes BP, Drucker DB, Zaia AA, Ferraz CC, Souza-Filho FJ (2004) Antimicrobial susceptibility of Enterococcus faecalis isolated from canals of root filled teeth with periapical lesions. Int Endod J 37:756–763

    Article  PubMed  Google Scholar 

  26. Sedgley CM, Molander A, Flannagan SE, Nagel AC, Appelbe OK, Clewell DB, Dahlén G (2005) Virulence, phenotype and genotype characteristics of endodontic Enterococcus spp. Oral Microbiol Immunol 20:10–19

    Article  PubMed  Google Scholar 

  27. Demo M (1996) Caracterizacion y estudios de patogenicidad de cepas dela genero Staphylococcus asiladas de leches mastiticas. Instituto de Microbiologia, Universidad Nacional de Rio Cuarto, Argentina.

  28. Marra A, Dib-Hajj F, Lamb L, Kaczmarek F, Shang W, Beckius G, Milici AJ, Medina I, Gootz TD (2007) Enterococcal virulence determinants may be involved in resistance to clinical therapy. Diagn Microbiol Infect Dis 58:59–65

    Article  PubMed  Google Scholar 

  29. Dunavant TR, Regan JD, Glickman GN, Solomon ES, Honeyman AL (2006) Comparative evaluation of endodontic irrigants against Enterococcus faecalis biofilms. J Endod 32:527–531

    Article  PubMed  Google Scholar 

  30. Pinheiro ET, Gomes BP, Ferraz CC, Sousa EL, Teixeira FB, Souza-Filho FJ (2003) Microorganisms from canals of root-filled teeth with periapical lesions. Int Endod J 36:1–11

    Article  PubMed  Google Scholar 

  31. Peciuliene V, Reynaud AH, Balciuniene I, Haapasalo M (2001) Isolation of yeasts and enteric bacteria in root-filled teeth with chronic apical periodontitis. Int Endod J 34:429–434

    Article  PubMed  Google Scholar 

  32. Orstavik D, Haapasalo M (1990) Disinfection by endodontic irrigants and dressings of experimentally infected dentinal tubules. Endod Dent Traumatol 6:142–149

    Article  PubMed  Google Scholar 

  33. Distel JW, Hatton JF, Gillespie MJ (2002) Biofilm formation in medicated root canals. J Endod 28:689–693

    Article  PubMed  Google Scholar 

  34. Ran S, He Z, Liang J (2013) Survival of Enterococcus faecalis during alkaline stress: changes in morphology, ultrastructure, physiochemical properties of the cell wall and specific gene transcripts. Arch Oral Biol 58:1667–1676

    Article  PubMed  Google Scholar 

  35. Strateva T, Atanasova D, Savov E, Petrova G, Mitov I (2016) Incidence of virulence determinants in clinical Enterococcus faecalis and Enterococcus faecium isolates collected in Bulgaria. Braz J Infect Dis 20:127–133

    Article  PubMed  Google Scholar 

  36. Lowe AM, Lambert PA, Smith AW (1995) Cloning of an Enterococcus faecalis endocarditis antigen: homology with adhesins from some oral streptococci. Infect Immun 63:703–706

    Article  PubMed  PubMed Central  Google Scholar 

  37. Linde A, Goldberg M (1993) Dentinogenesis. Crit Rev Oral Biol Med 4:679–728

    Article  PubMed  Google Scholar 

  38. Zhu X, Wang Q, Zhang C, Cheung GS, Shen Y (2010) Prevalence, phenotype, and genotype of Enterococcus faecalis isolated from saliva and root canals in patients with persistent apical periodontitis. J Endod 36:1950–1955

    Article  PubMed  Google Scholar 

  39. Mohamed JA, Huang W, Nallapareddy SR, Teng F (2004) Murray BE (2004) Influence of origin of isolates, especially endocarditis isolates, and various genes on biofilm formation by Enterococcus faecalis. Infect Immun 72:3658–3663

    Article  PubMed  PubMed Central  Google Scholar 

  40. Di Rosa R, Creti R, Venditti M, D'Amelio R, Arciola CR, Montanaro L, Baldassarri L (2006) Relationship between biofilm formation, the enterococcal surface protein (Esp) and gelatinase in clinical isolates of Enterococcus faecalis and Enterococcus faecium. FEMS Microbiol Lett 256:145–150

    Article  PubMed  Google Scholar 

  41. Hill PA, Docherty AJ, Bottomley KM, O'Connell JP, Morphy JR, Reynolds JJ, Meikle MC (1995) Inhibition of bone resorption in vitro by selective inhibitors of gelatinase and collagenase. Biochem J 15:167–175

    Article  Google Scholar 

  42. Tang SS, Apisarnthanarak A, Hsu LY (2014) Mechanisms of beta-lactam antimicrobial resistance and epidemiology of major community- and healthcare-associated multidrug-resistant bacteria. Adv Drug Deliv Rev 78:3–13

    Article  PubMed  Google Scholar 

  43. Tavares W (2002) Problem Gram-positive bacteria: resistance in staphylococci, enterococci and pneumococci to antimicrobial drugs. Rev Soc Bras Med Trop 35:409

    Article  PubMed  Google Scholar 

Download references

Funding

This work was financially supported by the Brazilian agencies Sao Paulo Research Foundation (FAPESP, 2015/23479-5, 2017/16516-7; 2017/26973-6 e 2018/00587-5;), National Council for Scientific and Technological Development (CNPq 308162/2014-5, 303852/2019-4) and Coordination for the Improvement of Higher Education Personnel (CAPES, finance code 001).

Author information

Authors and Affiliations

  1. Department of Restorative Dentistry, Division of Endodontics, Piracicaba Dental School, State University of Campinas, Av Limeira 901, Bairro Areiao, Piracicaba, São Paulo, Brazil

    Priscila Amanda Francisco, Pedro Ivo da Graça Fagundes, João Carlos Lemes-Junior, Augusto Rodrigues Lima, Maicon Ricardo Zieberg Passini & Brenda P. F. A. Gomes

Authors
  1. Priscila Amanda Francisco
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Pedro Ivo da Graça Fagundes
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. João Carlos Lemes-Junior
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Augusto Rodrigues Lima
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Maicon Ricardo Zieberg Passini
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Brenda P. F. A. Gomes
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Brenda P. F. A. Gomes.

Ethics declarations

Ethical approval

All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.

Informed consent

Informed consent was obtained from all individual participants included in the study.

Conflict of interest

The authors declare that they have no conflict of interest.

Additional information

Publisher’s note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Francisco, P.A., Fagundes, P.I.d.G., Lemes-Junior, J.C. et al. Pathogenic potential of Enterococcus faecalis strains isolated from root canals after unsuccessful endodontic treatment. Clin Oral Invest 25, 5171–5179 (2021). https://doi.org/10.1007/s00784-021-03823-w

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00784-021-03823-w

Keywords

Search

Navigation