Advertisement

Clinical Oral Investigations

, Volume 20, Issue 7, pp 1733–1739 | Cite as

Effect of gaseous ozone on Enterococcus faecalis biofilm–an in vitro study

  • Tanja BochEmail author
  • Christian Tennert
  • Kirstin Vach
  • Ali Al-Ahmad
  • Elmar Hellwig
  • Olga Polydorou
Original Article

Abstract

Objectives

The aim of this study was to evaluate the antimicrobial effect of gaseous ozone compared to conventional methods against Enterococcus faecalis.

Materials and methods

One hundred twenty-five teeth were infected by E. faecalis and were incubated for 72 h to form biofilm. Teeth were distributed among five groups. In the first group, ozone was used; in the second group, teeth were rinsed with 20 % ethylenediaminetetraacetic acid (EDTA); in the third group, with 3 % sodium hypochlorite (NaOCl). Group 4 combined 20 % EDTA with ozone. NaOCl and ozone were combined in group 5. After treatment, the samples with paper points were taken, followed by dentin samples taken with K-file, and cultured for 24 h. Then bacterial colonies were counted.

Results

All treatments reduced significantly (p < 0.05) the bacteria. Paper points’ samples showed 85.38 % reduction after ozone. The highest reduction was observed in NaOCl group (99.98 %). EDTA reduced bacteria by 80.64 %. Combination of NaOCl and ozone eradicated 99.95 % of the bacteria. Combination of EDTA and ozone reduced E. faecalis up to 91.33 %. The dentin chips showed the following: the highest CFU counts were observed in EDTA group, followed by ozone and NaOCl group. The lowest CFU counts were found in NaOCl-ozone group and EDTA-ozone group.

Conclusions

Ozone reduced E. faecalis, even organised in a biofilm, however, lower than NaOCl. No treatment reduced totally the bacteria.

Clinical relevance

Used as an adjuvant, ozone can increase the efficacy of conventional rinsing like EDTA and presents an alternative treatment when NaOCl cannot be used e.g. in teeth with a wide-open apical foramen.

Keywords

Ozone Enterococcus faecalis Root canal treatment Endodontics Disinfection methods 

Notes

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflicts of interest.

Research involving human participants and/or animals

Ethical approval

All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards. This article does not contain any studies with animals performed by any of the authors.

References

  1. 1.
    Byström A, Sundqvist G (1983) Bacteriologic evaluation of the effect of 0.5 percent sodium hypochlorite in endodontic therapy. Oral Surg Oral Med Oral Pathol 55:307–312CrossRefPubMedGoogle Scholar
  2. 2.
    Almyroudi A, Mackenzie D, McHugh S, Saunders WP (2002) The effectiveness of various disinfectants used as endodontic intracanal medications: an in vitro study. J Endod 28:163–167CrossRefPubMedGoogle Scholar
  3. 3.
    Wu MK, Wesselink PR (2001) A primary observation on the preparation and obturation of oval canals. Int Endod J 34:137–141CrossRefPubMedGoogle Scholar
  4. 4.
    Radcliffe CE, Potouridou L, Qureshi R, Habahbeh N, Qualtrough A, Worthington H, Drucker DB (2004) Antimicrobial activity of varying concentrations of sodium hypochlorite on the endodontic microorganisms Actinomyces israelii, A. naeslundii, Candida albicans and Enterococcus faecalis. Int Endod J 37:438–446CrossRefPubMedGoogle Scholar
  5. 5.
    Vianna ME, Gomes BP, Berber VB, Zaia AA, Ferraz CC, De Souza-Filho FJ (2004) In vitro evaluation of the antimicrobial activity of chlorhexidine and sodium hypochlorite. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 97:79–84CrossRefPubMedGoogle Scholar
  6. 6.
    Zehnder M (2006) Root canal irrigants. J Endod 32:389–398CrossRefPubMedGoogle Scholar
  7. 7.
    Narayanan L, Vaishnavi C (2010) Endodontic microbiology. J Conserv Dent 13:233–239CrossRefPubMedPubMedCentralGoogle Scholar
  8. 8.
    Rôças IN, Siqueira Jr JF, Santos KR (2004) Association of Enterococcus faecalis with different forms of periradicular diseases. J Endod 30:315–320CrossRefPubMedGoogle Scholar
  9. 9.
    Schirrmeister JF, Liebenow AL, Pelz K, Wittmer A, Serr A, Hellwig E, Al-Ahmad A (2009) New bacterial compositions in root-filled teeth with periradicular lesions. J Endod 35:169–174CrossRefPubMedGoogle Scholar
  10. 10.
    Sundqvist G, Figdor D, Persson S, Sjögren 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 85:87–94CrossRefGoogle Scholar
  11. 11.
    Garcez AS, Nunez SC, Hamblim MR, Suzuki H, Ribeiro MS (2010) Photodynamic therapy associated with conventional endodontic treatment in patients with antibiotic-resistant microflora: a preliminary report. J Endod 36:1463–1466CrossRefPubMedGoogle Scholar
  12. 12.
    Kishen A, Upadya M, Tegos GP, Hamblin MR (2010) Efflux pump inhibitor potentiates antimicrobial photodynamic inactivation of Enterococcus faecalis biofilm. Photochem Photobiol 86:1343–1349CrossRefPubMedPubMedCentralGoogle Scholar
  13. 13.
    Nagayoshi M, Kitamura C, Fukuizumi T, Nishihara T, Terashita M (2004) Antimicrobial effect of ozonated water on bacteria invading dentinal tubules. J Endod 30:778–781CrossRefPubMedGoogle Scholar
  14. 14.
    Ng R, Singh F, Papamanou DA, et al. (2011) Endodontic photodynamic therapy ex vivo. J Endod 37:217–222CrossRefPubMedPubMedCentralGoogle Scholar
  15. 15.
    Noiri Y, Katsumoto T, Azakami H, Ebisu S (2008) Effects of Er:YAG laser irradiation on biofilm-forming bacteria associated with endodontic pathogens in vitro. J Endod 34:826–829CrossRefPubMedGoogle Scholar
  16. 16.
    Nunes MR, Mello I, Franco GC, de Medeiros JM, Dos Santos SS, Habitante SM, Lage-Marques JL, Raldi DP (2011) Effectiveness of photodynamic therapy against Enterococcus faecalis, with and without the use of an intracanal optical fiber: an in vitro study. Photomed Laser Surg 29:803–808CrossRefPubMedGoogle Scholar
  17. 17.
    Sennhenn-Kirchner S, Schwarz P, Schliephake H, Konietschke F, Brunner E, Borg-von Zepelin M (2009) Decontamination efficacy of erbium:yttrium-aluminium-garnet and diode laser light on oral Candida albicans isolates of a 5-day in vitro biofilm model. Lasers Med Sci 24:313–320CrossRefPubMedGoogle Scholar
  18. 18.
    Fimple JL, Fontana CR, Foschi F, et al. (2008) Photodynamic treatment of endodontic polymicrobial infection in vitro. J Endod 34:728–734CrossRefPubMedPubMedCentralGoogle Scholar
  19. 19.
    Meire MA, Coenye T, Nelis HJ, De Moor RJ (2012) In vitro inactivation of endodontic pathogens with Nd:YAG and Er:YAG lasers. Lasers Med Sci 27:695–701CrossRefPubMedGoogle Scholar
  20. 20.
    Silva LA, Novaes Jr AB, de Oliveira RR, Nelson-Filho P, Santamaria Jr M, Silva RA (2012) Antimicrobial photodynamic therapy for the treatment of teeth with apical periodontitis: a histopathological evaluation. J Endod 38:360–366CrossRefPubMedGoogle Scholar
  21. 21.
    Holmes J (2003) Clinical reversal of root caries using ozone, double-blind, randomised, controlled 18-month trial. Gerodontology 20:106–114CrossRefPubMedGoogle Scholar
  22. 22.
    Huth KC, Paschos E, Brand K, Hickel R (2005) Effect of ozone on non-cavitated fissure carious lesions in permanent molars. A controlled prospective clinical study. Am J Dent 18:223–228PubMedGoogle Scholar
  23. 23.
    Hems RS, Gulabivala K, Ng YL, Ready D, Spratt DA (2005) An in vitro evaluation of the ability of ozone to kill a strain of Enterococcus faecalis. Int Endod J 38:22–29CrossRefPubMedGoogle Scholar
  24. 24.
    Ureyen Kaya B, Kececi AD, Guldas HE, et al. (2014) Efficacy of endodontic applications of ozone and low-temperature atmospheric pressure plasma on root canals infected with Enterococcus faecalis. Lett Appl Microbiol 58:8–15CrossRefPubMedGoogle Scholar
  25. 25.
    Zan R, Hubbezoglu I, Sumer Z, Tunc T, Tanalp J (2013) Antibacterial effects of two different types of laser and aqueous ozone against Enterococcus faecalis in root canals. Photomed Laser Surg 31:150–154CrossRefPubMedGoogle Scholar
  26. 26.
    Huth KC, Quirling M, Maier S, Kamereck K, Alkhayer M, Paschos E, Welsch U, Miethke T, Brand K, Hickel R (2009) Effectiveness of ozone against endodontopathogenic microorganisms in a root canal biofilm model. Int Endod J 42:3–13CrossRefPubMedGoogle Scholar
  27. 27.
    Cheng X, Guan S, Lu H, Zhao C, Chen X, Li N, Bai Q, Tian Y, Yu Q (2012) Evaluation of the bactericidal effect of Nd:YAG, Er:YAG, Er,Cr:YSGG laser radiation, and antimicrobial photodynamic therapy (aPDT) in experimentally infected root canals. Lasers Surg Med 44:824–831CrossRefPubMedGoogle Scholar
  28. 28.
    Tennert C, Drews AM, Walther V, Altenburger MJ, Karygianni L, Wrbas KT, Hellwig E, Al-Ahmad A (2015) Ultrasonic activation and chemical modification of photosensitizers enhances the effects of photodynamic therapy against Enterococcus faecalis root-canal isolates. Photodiagnosis Photodyn Ther 12(2):244–251Google Scholar
  29. 29.
    Bergmans L, Moisiadis P, Huybrechts B, Van Meerbeek B, Quirynen M, Lambrechts P (2008) Effect of photo-activated disinfection on endodontic pathogens ex vivo. Int Endod J 41:227–239CrossRefPubMedGoogle Scholar
  30. 30.
    Foschi F, Fontana CR, Ruggiero K, Riahi R, Vera A, Doukas AG, Pagonis TC, Kent R, Stashenko PP, Soukos NS (2007) Photodynamic inactivation of Enterococcus faecalis in dental root canals in vitro. Lasers Surg Med 39:782–787CrossRefPubMedGoogle Scholar
  31. 31.
    Kayaoglu G, Ömürlü H, Akca G, Gürel M, Gencay Ö, Sorkun K, Salih B (2011) Antibacterial activity of Propolis versus conventional endodontic disinfectants against Enterococcus faecalis in infected dentinal tubules. J Endod 37:376–381CrossRefPubMedGoogle Scholar
  32. 32.
    Kuştarci A, Sumer Z, Altunbas D, Kosum S (2009) Bactericidal effect of KTP laser irradiation against Enterococcus faecalis compared with gaseous ozone: an ex vivo study. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 107:73–79CrossRefGoogle Scholar
  33. 33.
    Siqueira Jr JF, Rocas IN (2004) Polymerase chain reaction-based analysis of microorganisms associated with failed endodontic treatment. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 97:85–94CrossRefPubMedGoogle Scholar
  34. 34.
    Rich RL, Kreikemeyer B, Owens RT, La Brenz S, Narayana SV, Weinstock GM, Murray BE, Höök M (1999) Ace is a collagen-binding MSCRAMM from Enterococcus facealis. J Biol Chem 274:26939–26945CrossRefPubMedGoogle Scholar
  35. 35.
    Rozdzinski E, Marre R, Susa M, Wirth R, Muscholl-Silberhorn A (2001) Aggregation substance-mediated adherence of Enterococcus faecalis to immobilized extracellular matrix proteins. Microb Pathog 30:211–220CrossRefPubMedGoogle Scholar
  36. 36.
    Distel JW, Hatton JF, Gillespie MJ (2002) Biofilm formation in medicated root canals. J Endod 28:689–693CrossRefPubMedGoogle Scholar
  37. 37.
    Hancock HH, Sigurdsson A, Trope M, Moiseiwitsch J (2001) Bacteria isolated after unsuccessful endodontic treatment in a North American population. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 91:579–586CrossRefPubMedGoogle Scholar
  38. 38.
    Atila-Pektas B, Yurdakul P, Gülmez D, Görduysus O (2013) Antimicrobial effects of root canal medicaments against Enterococcus faecalis and Streptococcus mutans. Int Endod J 46:413–418CrossRefPubMedGoogle Scholar
  39. 39.
    Dametto FR, Ferraz CC, Gomes BP, Zaia AA, Teixeira FB, De Souza-Filho FJ (2005) In vitro assessment of the immediate and prolonged antimicrobial action of chlorhexidine gel as an endodontic irrigant against Enterococcus faecalis. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 99:768–772CrossRefPubMedGoogle Scholar
  40. 40.
    Tennert C, Fuhrmann M, Wittmer A, Karygianni L, Altenburger MJ, Pelz K, Hellwig E, Al-Ahmad A (2014) New bacterial composition in primary and persistent/secondary endodontic infections with respect to clinical and radiographic findings. J Endod 40:670–677CrossRefPubMedGoogle Scholar
  41. 41.
    Schirrmeister JF, Liebenow AL, Braun G, Wittmer A, Hellwig E, Al-Ahmad A (2007) Detection and eradication of microorganisms in root-filled teeth associated with periradicular lesions: an in vivo study. J Endod 33:536–540CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2015

Authors and Affiliations

  • Tanja Boch
    • 1
    Email author
  • Christian Tennert
    • 1
  • Kirstin Vach
    • 2
  • Ali Al-Ahmad
    • 1
  • Elmar Hellwig
    • 1
  • Olga Polydorou
    • 1
  1. 1.Department of Operative Dentistry and PeriodontologyUniversity Medical Center FreiburgFreiburg i. BrGermany
  2. 2.Center for Medical Biometry and Medical Informatics, Institute for Medical Biometry and StatisticsMedical Center–University of FreiburgFreiburgGermany

Personalised recommendations