An in-vitro investigation of the antibacterial effect of nisin in root canals and canal wall radicular dentine

Abstract

Aim: To determine whether nisin, a bacteriocin, would be effective at killing Enterococcus faecalis and Streptococcus gordonii cells in solution and within the root canal system.

Methodology: Bacterial isolates of E. faecalis and S. gordonii were grown from glycerol stocks in closed tubes containing BHY broth at 37 °C. The minimum bactericidal concentration (MBC) of nisin for both bacterial species was determined by a microdilution method. Extracted human teeth were decoronated to produce roots of equal length with a single canal and divided into six groups of 10 roots. The canals were prepared to a master apical size 30 file using 0.04 taper Ni-Ti rotary instruments. Bacterial samples of each species were inoculated into three groups of prepared roots and incubated in closed tubes at 37 °C for 21 days. The root canals in each group were then medicated with water (control), calcium hydroxide powder mixed with sterile water [Ca(OH)2], or nisin and incubated for a further 7 days. Rotary Ni-Ti files were used to take radicular dentine samples from the walls of each canal which were then incubated in BHY broth for 24 h. Optical density (OD600) readings were taken as a measure of bacterial growth.

Results: The MBC of nisin for E. faecalis and S. gordonii was 70 and 20 mg mL−1 respectively. Calcium hydroxide and nisin medication eradicated infection within the root canal while cells remained viable in the control group. Mean optical density (OD600) readings from canal wall dentine shavings infected with E. faecalis were 1.32 ± 0.98, 0.73 ± 0.27 and 0.69 ± 0.38 for the control, Ca(OH)2 and nisin samples respectively. Corresponding mean readings for S. gordonii were 1.19 ± 0.18, 0.73 ± 0.15 and 0.60 ± 0.29. The Ca(OH)2 and nisin group readings were significantly (P < 0.01) lower than the control for each species as tested by Student's t-test and Mann–Whitney U statistical analysis. Values for Ca(OH)2 and nisin were not significantly (P > 0.01) different.

Conclusion: Nisin was effective at eradicating E. faecalis and S. gordonii cells in pure culture and was comparable with Ca(OH)2 in the elimination of these species from within the root canal system.