Comparative analysis of Enterococcus faecalis biofilm formation on different substrates

Juliane Maria Guerreiro-Tanomaru, Norberto Batista De Faria-Júnior, Marco Antônio Húngaro Duarte, Ronald Ordinola-Zapata, Márcia Sirlene Zardin Graeff, Mário Tanomaru-Filho

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56 Scopus citations


Introduction: The aim of this study was to compare Enterococcus faecalis biofilm formation on different substrates. Methods: Cell culture plates containing growth medium and E. faecalis (ATCC 29212) were used to grow biofilm on bovine dentin, gutta-percha, hydroxyapatite, or bovine bone. Substrates were incubated at 37°C for 14 or 21 days, and the medium was changed every 48 hours. After the growth induction periods, specimens (n = 5 per group and per induction period) were stained by using Live/Dead, and the images were analyzed under a confocal microscope. The total biovolume (μm3), live bacteria biovolume (μm3), and substrate coverage (%) were quantified by using the BioImage-L software. Results obtained were analyzed by nonparametric tests (P =.05). Results: Biofilm formation was observed in all groups. Gutta-percha had the lowest total biovolume at 14 days (P <.05) and hydroxyapatite the highest at 21 days (P <.05). No significant difference was observed in green biovolume at 14 days. At 21 days, however, hydroxyapatite had the highest volume (P <.05). The percentages of coverage were similar among all substrates at 21 days (P >.05), but at 14 days, bovine bone presented the highest coverage (P <.05). Conclusions: E. faecalis was capable of forming biofilm on all substrates during both growth periods; hydroxyapatite presented the highest rates of biofilm formation. The type of substrate influenced the biofilm characteristics, according to the parameters evaluated.

Original languageEnglish (US)
Pages (from-to)346-350
Number of pages5
JournalJournal of Endodontics
Issue number3
StatePublished - Mar 2013
Externally publishedYes


  • Biofilm
  • Enterococcus faecalis
  • confocal microscopy


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