Elsevier

Journal of Endodontics

Volume 38, Issue 8, August 2012, Pages 1114-1117
Journal of Endodontics

Basic Research
Efficacy of Berberine, an Antimicrobial Plant Alkaloid, as an Endodontic Irrigant against a Mixed-culture Biofilm in an In Vitro Tooth Model

https://doi.org/10.1016/j.joen.2012.04.023Get rights and content

Abstract

Introduction

Berberine, a plant alkaloid isolated from many medicinal plants, has shown antimicrobial activity against selected oral pathogens. The purpose of this investigation was to evaluate the antimicrobial efficacy of berberine solution against selected endodontic pathogens using a multispecies biofilm tooth model.

Methods

The bacterial species used in the multispecies biofilm tooth model were Fusobacterium nucleatum, Enterococcus faecalis, and Prevotella intermedia. Extracted human anterior teeth were collected and standardized to a length of 14.0 mm. Teeth were cultured in Schaedler broth with the 3 test bacteria strains for 21 days and then randomly assigned to 6 treatment groups (ie, sterile saline, 5.25% NaOCl, 2% chlorhexidine [CHX], 1% CHX, 2 mg/mL berberine, and 1 mg/mL berberine plus 1% CHX). The teeth were instrumented to size 35/.06 and irrigated with 6 mL irrigant for 2 minutes. Surviving bacteria were sampled before and after instrumentation. Data were analyzed using analysis of variance (P < .05) followed by the Scheffé test.

Results

The minimal inhibitory concentration of berberine against F. nucleatum, P. intermedia, and E. faecalis was 31.25 μg/mL, 3.8 μg/mL, and 500 μg/mL, respectively. Instrumentation and irrigation resulted in 99% bacterial reduction in all groups. All tested solutions resulted in a statistically significant reduction in bacteria when compared with the saline control. When used alone, berberine (2 mg/mL) was less effective than the other test irrigants. However, when combined with 1% CHX, berberine (2 mg/mL) was comparable in bactericidal activity with 5.25% NaOCl, 2% CHX, and 1% CHX (Table 2).

Conclusions

Berberine was more effective than saline as an endodontic irrigant against selected endodontic pathogens in vitro and, when combined with CHX, was comparable with NaOCl in its bactericidal efficacy.

Section snippets

Antibacterial Activity and Bacterial Growth Assay

Minimal inhibitory concentrations (MICs) were determined using a microdilution method in microtiter plates (24). Test bacteria included E. faecalis American Type Culture Collection (ATCC) 29212, P. intermedia ATCC 25611, and F. nucleatum ATCC 10953. The final inoculum concentrations were 5 × 105 colony-forming units (CFU)/mL for E. faecalis and 5 × 106 CFU/mL for F. nucleatum or P. intermedia. Controls included an inoculated growth medium without test compounds, and sample blanks contained

Results

The MICs of the test agents against tested bacteria strains are listed in Table 1. All test agents inhibited growth of the selected bacteria. BBr inhibited the in vitro growth of F. nucleatum (MIC = 31.25 μg/mL), P. intermedia (MIC = 3.8 μg/mL), and E. faecalis (MIC = 500 μg/mL) in brain-heart infusion medium. E. faecalis was less sensitive to BBr when compared with the other 2 test bacteria.

Intergroup analysis of S1 (bacterial loads in the canals before instrumentation and irrigation) samples

Discussion

The polymicrobial nature of endodontic infections is well established (25). The intracanal microbiota in endodontically infected teeth exists both as a loose collection and as biofilm structures, consisting of cocci, rods, and filamentous bacteria (26). Many in vitro studies used planktonic cultures (bacteria in suspension) or monospecies biofilms 12, 27. In the current study, we established an in vitro mixed-culture biofilm model system to investigate the bactericidal activity of selected

Acknowledgments

The authors deny any conflicts of interest related to this study.

References (32)

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This study was supported by an American Association of Endodontists Foundation research grant and the Departments of Endodontics and Pediatric Dentistry, University of Illinois at Chicago.

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