Comparison of the use of d-enantiomeric and l-enantiomeric antimicrobial peptides incorporated in a calcium-chelating irrigant against Enterococcus faecalis root canal wall biofilms.

OBJECTIVES: To compare the anti-biofilm efficacy of two antimicrobial peptides (AMPs), 1018 and DJK-5, in disrupting canal wall biofilms in the isthmus, canal and dentinal tubules of single-rooted maxillary premolars. METHODS: Enterococcus faecalis single-species biofilms were formed in-situ in the root canal system of the premolars (n = 91). Confocal laser scanning microscopy, bacterial sampling, colony-forming unit counting, XTT assay, lactate dehydrogenase assay and phenol-sulphuric acid method were used to identify the anti-biofilm efficacy of both AMPs and their influence on bacterial metabolic activity. RESULTS: Both AMPs disrupted in-situ E. faecalis biofilms and altered their metabolic activity. At 20 µg/mL, the d-enantiomeric AMP DJK-5 killed 55.5 %, 57.3 % and 55.8 % of biofilm bacteria in the isthmus, canal and dentinal tubules, respectively, in 1 min. In contrast, the l-enantiomeric AMP 1018 only eradicated 25.6 %, 25.5 % and 27.5 % of biofilm bacteria in the isthmus, canal and dentinal tubules, respectively, within the same time. Anti-biofilm efficacy of the root canal irrigants tested were in the order: 6 % NaOCl > 20 µg/mL DJK-5 > 10 µg/mL DJK-5 > 20 µg/mL 1018 > 10 µg/mL 1018 > 0.9 % NaCl. CONCLUSIONS: The present results are confirmatory of previous studies, in that d-enantiomeric AMPs exhibit more potent antibacterial properties than l-enantiomeric AMPs against E. faecalis biofilms within the canal space. Nevertheless, the potency of both AMPs are concentration-dependent. Incorporation of these agents into EDTA, a non-antibacterial calcium-chelating irrigant for removal of the inorganic component of the canal space debris, does not reduce the efficacy of either AMP. CLINICAL SIGNIFICANCE: The present study provides the proof of concept that incorporation of an antimicrobial peptide into a calcium-chelating root canal irrigant enhances the disinfection of intratubular single-species biofilms during smear layer and smear plug removal.

Anti-biofilm efficacy of root canal irrigants against in-situ Enterococcus faecalis biofilms in root canals, isthmuses and dentinal tubules.

OBJECTIVE: To investigate the anti-biofilm efficacy of root canal irrigants in canal spaces, isthmi and dentinal tubules of root canals ex vivo. METHODS: Fifty-one single-rooted premolars, each containing an isthmus, were instrumented, autoclaved and inoculated with Enterococcus faecalis for 4 weeks. One specimen was sectioned for bacteria-specific staining to confirm the presence of biofilms using light microscopiy. The remaining specimens were randomly divided to five groups: (1) 0.9% NaCl, (2) SilverSol/H2O2, (3) HYBENX, (4) QMix 2 in1, (5) 6% NaOCl. Bacterial sampling was performed before (S1) and after (S2) canal irrigation. Diluted bacteria suspension was cultured for 48 h for counting the colony forming units (CFU). Percentages of dead bacteria and biofilm thickness were evaluated by confocal laser scanning microscopy (CLSM). Metabolic activity, lactic acid and polysaccharide synthesis of E. faecalis derived from S2 samples were analysed. RESULTS: The percentages of dead bacteria were significantly affected by the factor "irrigant" (p < 0.001) and the factor "location" (p = 0.017). The percentages of dead bacteria in the isthmi and canals were both in the ordor: NaCl < SilverSol/H2O2 < HYBENX < QMix 2 in1 < NaOCl (p < 0.05). Only 6% NaOCl disrupted biofilms and significantly reduced their thickness. The CFU, metabolic activity, polysaccharide and lactic acid production of E. faecalis were all reduced by the disinfecting solutions. CONCLUSIONS: SilverSol/H2O2 and HYBENX were less adept than QMix 2 in1 at killing biofilm bacteria in root canals. None of these antibacterial irrigants were effective, compared with 6% NaOCl, in disrupting biofilms. CLINICAL SIGNIFICANCE: There is advantage in using HYBENX or QMix 2 in1 to kill intratubular bacteria biofilms because of their capability in removing the inorganic component of the smear layer. SilverSol/H2O2 requires extra time to eradicate intratubular biofilms upon removal of the organic and inorganic components of the smear layer by other root canal irrigants.