Possible role of the adhesin ace and collagen adherence in conveying resistance to disinfectants on Enterococcus faecalis

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Abstract

Introduction

This study aimed to evaluate whether the presence of the ace gene and Ace-mediated binding to collagen confers on Enterococcus faecalis resistance against common endodontic disinfectants.

Methods

Isogenic strains of E. faecalis: OG1RF (wild-type) and TX5256 (ace insertion mutant of OG1RF) were grown in brain–heart infusion broth at 46°C overnight. Standardized bacterial suspensions were pretreated for 1 h either with acid-soluble collagen or acidified phosphate-buffered saline (ac-PBS). Bacteria were challenged with chlorhexidine digluconate (CHX), iodine potassium-iodide (IKI), sodium hypochlorite (NaOCl), and calcium hydroxide [Ca(OH)2]. Samples were removed at 1, 3, and 6 h, and cultured on Todd–Hewitt agar plates. Colonies were counted, the absolute values were log transformed, and the data were statistically analyzed using Fisher's least significant differences test and t-test.

Results

OG1RF was more resistant than TX5256 to IKI, NaOCl, and Ca(OH)2 (P< 0.05). Collagen-exposed OG1RF was more resistant than the ac-PBS-pretreated OG1RF against CHX at 3 h and against IKI at 1 h (P < 0.05); no significant difference was found against NaOCl. As expected, the ace mutant strain, TX5256, pretreated with collagen or ac-PBS did not differ significantly in viability when challenged with CHX, IKI, and NaOCl. An unexpected result was found for Ca(OH)2: collagen-pretreated OG1RF and TX5256 were both more susceptible than ac-PBS-pretreated OG1RF and TX5256, respectively (P < 0.05).

Conclusion

The presence of the ace gene confers resistance against IKI, NaOCl, and Ca(OH)2 on E. faecalis. Exposure to collagen makes the wild-type bacterium more resistant against CHX and IKI; however, exposure to collagen apparently decreases resistance to Ca(OH)2.

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