The presence of smear layer affects the antimicrobial action of root canal sealers.

AIM: To assess the chemical and microstructural characteristics of dentine after the use of two irrigation protocols and correlate this with the antimicrobial properties of hydraulic calcium silicate cement (HCSC) sealers and changes to the dentine structure/chemistry after sealer placement. METHODOLOGY: Two irrigation protocols - Protocol A using 2% NaOCl used 5 mL/5 min and Protocol B with 2% NaOCl (5 mL/5 min) followed by 17% EDTA (5 mL/3 min) - were used to prepare dentine. The chemical and microstructural changes following irrigation were assessed by scanning electron microscopy (SEM), energy dispersive spectroscopy (EDS) and Fourier transform infrared (FT-IR) spectroscopy (n = 5) on dentine obtained from the mid-root and coronal parts of extracted human teeth. Four sealers (AH Plus, BioRoot, MTA Fillapex, TotalFill) were characterized by SEM/EDS (n = 3). The ability of the sealers to eradicate intratubular Enterococcus faecalis biofilms was assessed by live/dead dye and confocal laser scanning microscopy to measure the percentage of living cells. The effect of combined irrigation and root filling on dentine was assessed by SEM and EDS analysis (n = 5). Statistical analysis was undertaken using one-way anova and a number of post hoc tests to detect intergroup differences. The F-test was used for comparison of variances in the microbiology testing. RESULTS: The use of NaOCl alone left the smear layer intact, with traces of chlorine remaining on dentine. The use of BioRoot sealer restored the calcium levels of dentine which had been depleted by the irrigation with EDTA. BioRoot exhibited antimicrobial properties against intratubular bacteria even in the presence of smear layer (Protocol A). Smear layer removal improved the bactericidal effect of all sealers and Ca2+ leaching. The use of a chelating agent was important for the intratubular sealer penetration for AH Plus but not the other sealers. CONCLUSION: The removal of smear layer was necessary for penetration of AH Plus into the dentinal tubules. BioRoot was a more effective sealer in reducing the bacterial load in the dentinal tubules than the other materials tested and the presence of smear layer did not affect its activity.

Analysis of the effects of several decalcifying agents alone and in combination with sodium hypochlorite on the chemical composition of dentine.

AIM: To investigate the effects of several decalcifying agents alone and in combination with sodium hypochlorite (NaOCl) on the organic and inorganic components of dentine using attenuated total reflectance in Fourier transform infrared spectroscopy (ATR-FTIR). METHODOLOGY: Dentine slices from bovine teeth were submitted to (n = 5) the following: 0.9% saline, 9% and 18% etidronic acid (HEDP), 5% and 10% tetrasodium EDTA (EDTANa4 ), 17% trisodium EDTA (EDTAHNa3 ), and 0.5% and 2.0% peracetic acid (PAA) for 0.5-10 min; and to the combinations: G1 - mixture 5% NaOCl + 18% HEDP (5 and 10 min); G2 - mixture 5% NaOCl + 10% EDTANa4 (5 and 10 min); G2 - 2.5% NaOCl (5 min) + 17% EDTAHNa3 (1 min); G3 - 2.5% NaOCl (5 min) + 0.5% PAA (1 min); G4 - 2.5% NaOCl (5 min) + 9% HEDP (5 min). Specimens of G2, G3 and G4 received final flushes with 2.5% NaOCl for 0.5-10 min. Amide III/phosphate and carbonate/phosphate ratios of the spectra collected from the dentine specimens before and after immersion in the solutions were determined. Data were submitted to one-way repeated measures and one-way anova. RESULTS: For the same decalcifying agent, the higher the concentration and immersion time the greater the removal of phosphate, exposure of collagen matrix and consequently the increases in amide III/phosphate ratio. However, significant differences were found only between the two concentrations of PAA (P < 0.05). PAA caused greater increases in this ratio, followed by EDTAHNa3 , EDTANa4 and HEDP, and this order was retained in the combinations with NaOCl. This ratio was significantly reduced in G1 (P < 0.05) and not altered in G2 (P > 0.05). Due to collagen degradation, the amide III/phosphate ratio reduced significantly after the use of NaOCl in G3, G4 and G5 (P < 0.05). NaOCl required approximately 0.5 s to deproteinate the collagen matrix exposed after phosphate removal by EDTAHNa3 and PAA. The carbonate of dentine was removed more rapidly than phosphate by all decalcifying agents alone and in G3, G4 and G5. In the combinations with NaOCl, the last irrigant used defined the dentine amide III/phosphate and carbonate/phosphate ratios. CONCLUSIONS: HEDP and EDTANa4 caused minor whilst EDTAHNa3 and PAA caused greater demineralization of dentine; both effects were time and concentration dependent. NaOCl degraded the dentine organic matrix more rapidly when it was exposed. Combinations of NaOCl and decalcifying agents can be used to create dentine surfaces with varying compositions for interaction with endodontic sealers.

Mixture of alkaline tetrasodium EDTA with sodium hypochlorite promotes in vitro smear layer removal and organic matter dissolution during biomechanical preparation.

AIM: The aim of this study was to determine the following: (i) the quantity of free chlorine in mixtures of equal proportions of sodium hypochlorite (NaOCl) with trisodium ethylenediaminetetraacetic acid (EDTAHNa3 ) and alkaline tetrasodium ethylenediaminetetraacetic acid (EDTANa4 ); (ii) organic matter dissolution; and (iii) the time necessary to remove the smear layer by these irrigants alone and when mixed. METHODOLOGY: The solutions were mixed in a 1 : 1 ratio and then iodometrically titrated over time to determine the quantity of free available chlorine. The capability of organic matter dissolution by the solutions alone and the mixtures of irrigants was analysed by weighing bovine muscle tissue specimens before and after submission to the following groups (n = 10): G1 - 0.9% saline solution (control), G2 - 2.5% NaOCl, G3 - 17% EDTAHNa3 , G4 - 10% EDTANa4 , G5 - 20% EDTANa4 , G6 - 5% NaOCl + 17% EDTAHNa3 , G7 - 5% NaOCl + 10% EDTANa4 and G8 - 5% NaOCl + 20% EDTANa4 . The times necessary for smear layer removal were determinated on discs of bovine dentine with a standardized smear layer produced with SiC papers using a scanning electron microscope that did not require the samples to be sputter coated. The dentine discs were submitted to the same experimental groups previously described (n = 10) over several time periods, and the photomicrographs acquired were scored for the presence of smear layer. The parametric data of tissue dissolution were analysed using two-way anova and one-way anova with Tukey's post hoc tests (α < 0.05), whilst nonparametric data of smear layer removal were analysed by Friedman test (α < 0.05) and the Kruskal-Wallis test with Dunn's post hoc (α < 0.05). RESULTS: EDTAHNa3 caused an almost complete and immediate loss of free available chlorine from NaOCl, whilst EDTANa4 promoted a slow and concentrat-ion-dependent decline. The organic matter was not dissolved in the control group, EDTA groups or the mixture of NaOCl + 17% EDTAHNa3 group (P > 0.05). NaOCl alone and the associations of NaOCl + EDTANa4 dissolved tissue at all periods analysed (P < 0.05). The smear layer was not removed in the control and NaOCl groups (P > 0.05). The smear layer was removed at 1 min in the NaOCl + 17% EDTAHNa3 group (P < 0.05); 2 min in 17% EDTAHNa3 group (P < 0.05); and 5 min in 10% EDTANa4 , 20% EDTANa4 , 5% NaOCl + 10% EDTANa4 and 5% NaOCl + 20% EDTANa4 groups (P < 0.05). CONCLUSIONS: Alkaline EDTANa4 was slower in removing the smear layer than EDTAHNa3 , but when mixed with NaOCl during biomechanical canal preparation promoted organic matter dissolution and smear layer removal simultaneously. However, the mixing of NaOCl and EDTANa4 should be performed immediately before use to prevent the reduction of free available chlorine.