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



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.


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.


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.


Ozone Enterococcus faecalis Root canal treatment Endodontics Disinfection methods 


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.


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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

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