In vitro antimicrobial activity of several concentrations of sodium hypochlorite and chlorhexidine gluconate in the elimination of Enterococcus faecalis

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Abstract
The aim of this study was to assess, in vitro, the effectiveness of several concentrations of NaOCl (0.5%, 1%, 2.5%, 4% and 5.25%) and two forms of chlorhexidine gluconate (gel and liquid) in three concentrations (0.2%, 1% and 2%) in the elimination of E. faecalis. A broth dilution test using 24-well cell culture plates was performed and the time taken for the irrigants to kill bacterial cells was recorded. Isolated 24 h colonies of pure cultures of E. faecalis grown on 10% sheep blood plus Brain Heart Infusion (BHI) agar plates were suspended in sterile 0.85% NaCl solution. The cell suspension was adjusted spectrophotometrically to match the turbidity of a McFarland 0.5 scale. One mL of each tested substance was placed on the bottom of wells of 24-well cell culture plates (Corning, NY), including the control group (sterile saline). Six wells were used for each time period and irrigant concentration. Two mL of the bacterial suspension were ultrasonically mixed for 10 s with the irrigants and placed in contact with them for 10, 30, and 45 s; 1, 3, 5, 10, 20, and 30 min; and 1 and 2 h. After each period of time, 1 mL from each well was transferred to tubes containing 2 mL of freshly prepared BHI + neutralizers in order to prevent a residual action of the irrigants. All tubes were incubated at 37°C for 7 days. The tubes considered to have positive growth were those which presented medium turbidity during the incubation period. Data were analysed statistically by the Kruskal–Wallis test, with the level of significance set at P < 0.05. All irrigants were effective in killing E. faecalis, but at different times. Chlorhexidine in the liquid form at all concentrations tested (0.2%, 1% and 2%) and NaOCl (5.25%) were the most effective irrigants. However, the time required by 0.2% chlorhexidine liquid and 2% chlorhexidine gel to promote negative cultures was only 30 s and 1 min, respectively. Even though all tested irrigants possessed antibacterial activity, the time required to eliminate E. faecalis depended on the concentration and type of irrigant used.