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.

Effects of heat in the properties of NaOCl alone and mixed with etidronate and alkaline tetrasodium EDTA.

AIM: To assess the free available chlorine concentration (FAC), organic tissue dissolution and smear layer removal capacity of sodium hypochlorite (NaOCl) alone and when mixtured with etidronate (HEDP) and tetrasodium EDTA (Na4 EDTA), and heated to different temperatures. METHODOLOGY: Mixtures at 1 : 1 ratio of 5% NaOCl with distilled water (considered NaOCl alone), 18% HEDP or 10% Na4 EDTA were heated to 25 °C, 37 °C, 48 °C and 60 °C. The FAC in the mixtures was assessed at 5, 10, 20, 30, 60 and 120 min. Samples of bovine muscle tissue (n = 10) were prepared with similar size and weighed before and after 5, 10 and 15 min of immersion in the mixtures heated to the different temperatures to verify organic matter dissolution. The intergroup results were compared statistically using one-way analysis of variance (anova) and intragroup by two-way analysis of variance (anova), both followed by Tukey's multiple-comparison test (α < 0.01). Bovine dentine blocks (n = 10) were analysed by scanning electron microscopy before and after immersion in the mixtures, and the time taken to remove the smear layer from the surfaces of the samples was determined. The Friedman test was used to compare the scores of the same group (α < 0.01), and the Kruskal-Wallis test with Dunn's post hoc was used to compare the different groups (α < 0.01). Saline solution was used as a control in the experiments of tissue dissolution and smear layer removal, RESULTS: Heating NaOCl alone did not affect its FAC. The higher the temperature of the mixtures with the chelators, the lower the FAC. Organic tissue dissolution was improved by increases in temperature of NaOCl alone and its mixture with HEDP (P < 0.01); however, the mixture with Na4 EDTA had no improvement (P > 0.01). Smear layer removal by NaOCl alone was enhanced by heating resulting in lower scores in some samples and became more rapid in the mixtures with the chelators. The saline solution did not promote tissue dissolution nor smear layer removal (P > 0.01). CONCLUSION: In this laboratory study, heating NaOCl alone or when mixed with HEDP improved its capacity to dissolve organic matter and remove the smear layer. However, the mixture with HEDP required frequent refreshment to retain these effects when heated. Due to the acceleration in the reaction between the irrigants, very rapid reductions in the free available chlorine in the mixtures with Na4 EDTA heated to the different temperatures occurred.

A laboratory study of the scouting ability of two reciprocating glide path instruments in mesial root canals of extracted mandibular molars.

AIM: To evaluate in a laboratory setting the performance of two reciprocating glide path systems, WaveOne Gold Glider (WO) and R-Pilot (RP), to create a glide path in mesial root canals of mandibular molars and to assess the torsional resistance of instruments after performing the glide path. METHODOLOGY: A total of 60 mesial root canals of extracted human mandibular molars were divided into two groups (n = 30) according to the glide path system to be used. The data from the volume of each canal, acquired by microcomputed tomography (micro-CT), were validated statistically for the anatomical pairing of the groups. Preparation time, frequency in gaining apical patency, plastic deformation rate of instruments, and canal transportation and centring ability were recorded and compared statistically. The torsional fatigue of the instruments after use was also evaluated. Data were analysed using Fisher's exact test and Mann-Whitney U-test with a 5% significance level. RESULTS: No significant differences were found between groups regarding the time required to perform the glide path, the frequency distributions of the canals classified as patent and the instruments with plastic deformation after use (P > 0.05). No significant differences were found between groups regarding the degree of canal transportation and centring ability at the cervical, middle and apical thirds (P > 0.05). The RP groups had significantly greater maximum torsional strength values compared with the WO groups (P < 0.05). The used WO group had greater angular deflection to fracture when compared to the new WO group (P < 0.05). A significant difference was also found in the percentage of loss of angular deflection in a comparison of the WO group with the RP group (P < 0.05). CONCLUSIONS: The WO and RP instruments performed the same in terms of preparation time, plastic deformation, gaining apical patency, degree of canal transportation and centring ability. The RP instruments had greater torsional strength, less angular deflection and lower percentage of loss in angular deflection than the WO. The used WO group had the greatest angular deflection values.

Effectiveness of five instruments when removing calcium hydroxide paste from simulated internal root resorption cavities in extracted maxillary central incisors.

AIM: To evaluate the effectiveness of five instruments used for irrigant agitation during the removal of calcium hydroxide [Ca(OH)2 ] paste in simulated internal root resorption (IRR) cavities created in extracted maxillary central incisors. METHODOLOGY: Seventy maxillary central incisors with a single canal were selected. The canals were accessed and instrumented with Reciproc R50, then the roots were split in the bucco-lingual direction and the halves separated. Simulated IRR cavities were created, in both halves of the roots, 5 mm from the apex with a spherical bur. The specimens were reconstructed with cyanoacrylate glue and allocated into seven groups: negative control - no treatment; positive control - filled with Ca(OH)2 without performing any irrigation protocol; the other groups were divided according to the instrument used for irrigant agitation, namely: Ultrasonic, EndoActivator® , EDDY® , XP-endo® Finisher and XP-endo® Shaper. The specimens were cleaved and analysed using optical microscopy and scanning electron microscopy, to compare the Ca(OH)2 remnants between them. Then, the IRR cavities created by the burs were cleaned and subjected to a protocol of demineralization with 20% nitric acid, the roots reconstructed, and the irrigant agitation methods, as well as the microscopic analysis was repeated. Analysis of the images of Ca(OH)2 remaining in the simulated IRR cavities after irrigation was performed by two calibrated examiners based on a previously established scoring system. The data were statistically compared by Kruskal-Wallis test, Mann-Whitney U-test and Wilcoxon tests, with the significance level set at 5%. RESULTS: There was a significant difference in the effectiveness of the instruments in relation to the cavity creation method (bur vs. bur/acid) and evaluation method (optical microscopy and scanning electron microscopy) (P < 0.05). The XP-endo® Finisher and EDDY® groups were associated with significantly more effective removal of Ca(OH)2 when the IRR cavities were created using the acid protocol and analysed by scanning electron microscopy. CONCLUSION: None of the instruments tested were able to completely remove the Ca(OH)2 paste from the simulated IRR cavities; however, the EDDY® and XP-endo® Finisher removed more Ca(OH)2 in the bur/acid cavity creation method analysed by scanning electron microscopy.

Antimicrobial action of photodynamic therapy in root canals using LED curing light, curcumin and carbopol gel.

AIM: To evaluate the capacity of carbopol gel to maintain the intensity of a LED curing light (blueLED) along the length of prepared root canals in bovine teeth, and to assess the antimicrobial capacity of curcumin photoactivated by a LED curing light in the presence of carbopol gel. METHODOLOGY: Experiment 1: Eight straight roots of bovine incisors were standardized to a length of 15 mm, and the root canals instrumented up to a size 120 K-file. The LED curing light was irradiated inside the root canals using an aluminium collimator (1.5 mm in diameter) placed at the orifice (n = 8). Initially, the irradiation was performed in empty root canals and then repeated with the root canals filled with carbopol gel. Simple standardized photographs of the roots were taken with a digital camera in the mesial perspective during the irradiation procedure and the images analysed in OriginLab software to verify the light intensity along the length of the root. Experiment 2: Twenty dentine blocks were obtained from the cervical third of bovine incisors using a trephine bur. Biofilms were induced for 21 days on the blocks using Enterococcus faecalis (ATCC 4083) at 109 cells mL-1 . The blocks were treated according to the groups (n = 5): positive control; standard PDT (methylene blue + diode Laser); curcumin; LED curing light; and curcumin + LED curing light. After the treatment, the samples were dyed with Live/Dead BacLight Bacterial Viability solution and fluorescence images were obtained by Confocal Scanning Laser Microscopy (CSLM). Experiment 3: Thirty-two roots of bovine incisors were prepared as described in experiment 1. Their dentinal tubules were contaminated and the root canals treated according to the groups (n = 8): positive control; standard PDT; curcumin + LED curing light; curcumin + carbopol gel + LED curing light. The specimens were sectioned longitudinally and the split roots were treated with the Live/Dead dye to obtain fluorescence images by CSLM. All images were processed using BioImageL software to measure the percentage of viable bacteria and the data analysed statistically using the nonparametric Kruskal-Wallis test (α < 0.05). RESULTS: In Experiment 1, carbopol gel did not improve the intensity of LED light transmission along the root canal. In Experiment 2, a significant decrease (P < 0.05) in bacterial viability occurred in the following order: positive control < only LED curing light < only curcumin < curcumin + LED curing light = standard PDT; and in Experiment 3 positive control = curcumin + LED curing light ≤ curcumin + gel + LED curing light ≤ standard PDT. CONCLUSION: Similar disinfection effectiveness was obtained using curcumin + LED curing light and methylene blue + 660 nm LASER (standard PDT). The use of carbopol gel did not favour a greater transmission of LED light along the root canal and also resulted in less bacterial killing when used in endodontic PDT.

The ability of three nickel-titanium mechanized systems to negotiate and shape MB2 canals in extracted maxillary first molars: a micro-computed tomographic study.

AIM: To evaluate the influence of three engine-driven NiTi file systems manufactured from different NiTi alloys for the preparation of second mesiobuccal (MB2) canals in extracted maxillary first molars using a micro-computed tomographic (micro-CT) scanner. METHODOLOGY: Thirty maxillary molars with two canals in the mesiobuccal root were selected and randomly divided into three groups (n = 10): Reciproc [REC; size 25, .08 taper; VDW, Munich, Germany], ProDesign R [PDR; size 25, .06 taper; Easy, Belo Horizonte, Brazil] and Mtwo [MO; size 25, .06 taper; VDW, Munich, Germany]. Before and after root canal preparation of the MB2 canal, the teeth were scanned using a micro-CT to evaluate canal transportation, centring ability, dentine thickness and volume change. The working time to achieve working length was also evaluated. All parameters were compared statistically using the Kruskal-Wallis and Dunn test for multiple comparisons, with a significance level of 5%. RESULTS: There was no significant difference amongst the groups regarding canal transportation and centring ability (P > 0.05). However, the PDR size 25, .06 taper group had significantly lower canal volume and volume of dentine removal compared with a MO size 25, .06 taper and REC size 25, .08 taper (P < 0.05). A root perforation was detected in MO size 25, .06 taper and REC size 25, .08 taper groups, respectively. Regarding the working time, the PDR size 25, .06 taper required a significantly longer time to achieve working length than MO size 25, .06 taper and REC size 25, .08 taper (P < 0.05). CONCLUSIONS: All NiTi systems had similar canal transportation, centring ability and increase in apical volume after preparation of MB2 canals. However, the PDR size 25, .06 taper had less volume of dentine removal, absence of root canal perforation and required a longer time to accomplish the root canal preparation.

Effect of larger apical size on the quality of preparation in curved canals using reciprocating instruments with different heat thermal treatments.

AIM: To evaluate the influence of larger apical canal enlargement in curved canals using reciprocating systems subjected to various heat treatments. METHODOLOGY: Ninety mandibular premolars with root curvatures ranging from 20° to 30° were selected and scanned by microcomputed tomography (micro-CT) before and after root canal preparation with reciprocating systems (n = 30): Reciproc Blue (RB size 25, .08 taper and size 40, .06 taper; VDW, Munich, Germany), WaveOne Gold (WOG size 25, .07 taper and size 35, .06 taper; Dentsply Sirona, Ballaigues, Switzerland) and ProDesign R (PDR size 25, .06 taper and size 35, .05 taper; Easy Dental Equipment, Belo Horizonte, Brazil). Canal transportation, untouched areas, and apical and total root canal volumes were measured. Statistical analysis was performed with the nonparametric Kruskal-Wallis and Dunn's tests and a significance level set at 5%. RESULTS: The between-group comparison revealed no significant difference in untouched areas, canal transportation, and apical root canal volume among the groups (P > 0.05). However, WOG size 35, .06 taper was associated with a significant increase in the percentage of total canal volume in comparison to the PDR size 35, .05 taper (P < 0.05). The within-group comparison revealed a significant decrease in untouched areas, increase in apical and total root canal volume for all groups when using a larger instrument (P < 0.05). There was no significant difference in transportation among the groups and when a larger apical preparation was created (P > 0.05). CONCLUSIONS: Larger apical enlargement of curved canals was associated with a decrease in untouched areas, an increase in root canal volume and maintenance of canal trajectory. In addition, all systems were safe and provided similar root canal shapes.

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.

The effect of mixing method on tricalcium silicate-based cement.

AIM: To evaluate the effect of three methods of mixing on the physical and chemical properties of tricalcium silicate-based cements. METHODOLOGY: The materials evaluated were MTA Angelus and Portland cement with 20% zirconium oxide (PC-20-Zr). The cements were mixed using a 3 : 1 powder-to-liquid ratio. The mixing methods were manual (m), trituration (tr) and ultrasonic (us) activation. The materials were characterized by means of scanning electron microscope (SEM) and energy dispersive X-ray spectroscopy. Flowability was analysed according to ANSI/ADA 57/2012. Initial and final setting times were assessed following ASTM C266/08. Volume change was evaluated using a micro-CT volumetric method. Solubility was analysed according to ADA 57/2012. pH and calcium ion release were measured after 3, 24, 72 and 168 h. Statistical analysis was performed using two-way analysis of variance. The level of significance was set at P = 0.05. RESULTS: The SEM analysis revealed that ultrasonic activation was associated with a homogeneous distribution of particles. Flowability, volume change and initial setting time were not influenced by the mixing method (P > 0.05). Solubility was influenced by the mixing method (P < 0.05). For pH, at 168 h, significant differences were found between MTA-m and PC-20-Zr-m (P < 0.05). For calcium ion release, PC-20-Zr-tr had higher values than MTA-m at 3 h, and MTA-tr had higher values than PC-20-Zr-m at 168 h (P < 0.05). CONCLUSIONS: The ultrasonic and trituration methods led to higher calcium ion release and pH compared with manual mixing for all cements, whilst the ultrasonic method produced smaller particles for the PC-20-Zr cement. Flow, setting times and volume change were not influenced by the mixing method used; however, it did have an impact on solubility.

Antibacterial properties of silver nanoparticles as a root canal irrigant against Enterococcus faecalis biofilm and infected dentinal tubules.

AIM: To evaluate the antimicrobial action of an irrigant containing silver nanoparticles in an aqueous vehicle (AgNp), sodium hypochlorite and chlorhexidine against Enterococcus faecalis biofilm and infected dentinal tubules. METHODOLOGY: Bovine dentine blocks were used for E. faecalis biofilm development for 21 days and irrigated with 94 ppm AgNp solution, 2.5% NaOCl and 2% chlorhexidine for 5, 15 and 30 min. For infection of dentinal tubules with E. faecalis, dentine specimens from bovine incisors were submitted to a contamination protocol over 5 days, with eight centrifugation cycles on every alternate day, and irrigated with the same solutions and time intervals used for the biofilm. The specimens were stained with the Live/Dead technique and evaluated using a confocal laser scanning microscope (CLSM). The bioImage_L software was used for measurement of the total biovolume of biofilm in µm3 and percentage of viable bacteria (green cells) in biofilm and in dentinal tubules found after the irrigation. Statistical analyses were performed using Kruskal-Wallis and Dunn's tests for quantification of viable cells in biofilm, the Friedman test for comparisons of viable bacteria in dentinal tubules in different areas of the root canal and the Mann-Whitney U-test to compare the action of the irrigants between the two methods (P < 0.05). RESULTS: The AgNp solution eliminated fewer bacteria, but was able to dissolve more biofilm compared with chlorhexidine (P < 0.05). NaOCl had the greatest antimicrobial activity and biofilm dissolution capacity. AgNp solution had less antimicrobial action in infected dentinal tubules compared with NaOCl (P < 0.05). The AgNp solution after 5 min was more effective in eliminating planktonic bacteria in dentinal tubules than in biofilm, but at 30 min fewer viable bacteria were observed in the biofilm compared with intratubular dentine (P < 0.05). CONCLUSIONS: AgNp irrigant was not as effective against E. faecalis compared to solutions commonly used in root canal treatment. NaOCl is appropriate as an irrigant because it was effective in disrupting biofilm and in eliminating bacteria in biofilms and in dentinal tubules.

Evaluation of apical transportation and centring ability of five thermally treated NiTi rotary systems.

AIM: To evaluate apical transportation and centring ability during root canal preparation in mesial root canals of mandibular molars associated with ProTaper Gold (PTG), ProDesign S (PDS), Hyflex CM (HCM), Hyflex EDM and ProDesign Logic (PDL). METHODOLOGY: Sixty mandibular first molars with two separate canals in the mesial root were selected after root anatomy pairing by microcomputed tomography (microCT). The teeth were randomly divided into five groups (n = 24); the root canal volume was calculated to ensure sample homogeneity. All the root canals were prepared up to size 25 in accordance with the instructions of each rotary system manufacturer. After root canal preparation, the teeth were scanned by microCT to analyse apical transportation, root canal centralization and the pre- and post-preparation root canal volume at the apical and cervical levels. Kruskal-Wallis and Dunn tests were used for comparisons amongst groups for transportation values. For volume changes, the parametric ANOVA and Tukey's tests were used RESULTS: There were no significant differences in apical transportation amongst the rotary systems (P > 0.05). All the systems created apical transportation; values ranging from 0.031 mm (PDL) to 0.072 mm (PTG), and enlargements between 39% (HCM) and 91.1% (PDS) were observed. In relative to cervical transportation, significant differences were observed amongst the systems (P < 0.05). Mean transportation values between 0.07 mm (HCM) and 0.172 mm (PTG) were found, with enlargements between 35.4% (HCM) and 51.5% (PDS). CONCLUSION: All the thermally treated systems resulted in similar apical transportation. In the cervical region, the Hyflex CM and Prodesign Logic systems were associated with more centred preparations.

Torsional fatigue resistance of pathfinding instruments manufactured from several nickel-titanium alloys.

AIM: To evaluate the torsional properties of pathfinding nickel-titanium (NiTi) rotary instruments manufactured from several NiTi alloys, ProGlider (M-wire), Hyflex GPF (conventional NiTi Wire and controlled memory wire), Logic (conventional NiTi wire and controlled memory wire) and Mtwo (conventional NiTi wire). METHODOLOGY: A total of 56 NiTi instruments from Glidepath rotary systems (n = 8) were used: Logic (size 25, .01 taper), Logic CM (size 25, .01 taper), ProGlider (size 16, .02 taper), Hyflex GPF (size 15, .01 taper), Hyflex GPF CM (size 15, .02 taper; size 20, .02 taper) and Mtwo (size 10, .04 taper). The torsion tests were performed based on ISO 3630-1 (1992). Three millimetres of each instrument tip was clamped to a small load cell by a lever arm linked to the torsion axis. Data were analysed using a one-way analysis of variance (anova) and Tukey test with a significance level at a = 5%. RESULTS: The Logic size 25, .01 taper had significantly higher torsional strength values (P < 0.05). The ProGlider was significantly different when compared with Hyflex GPF size 15, .01 taper and size 15, .02 taper (P < 0.05). The Logic CM size 25, .01 taper had significantly higher torsional strength than Hyflex GPF size 15, .01 taper and size 15, .02 taper (P < 0.05). No difference was found amongst Mtwo size 10, .04 taper and Hyflex GPF groups (size 15, .01 taper; size 15, .02 taper; size 20, .02 taper). In relation to the angle of rotation, Logic CM size 25, .01 taper and Hyflex GPF size 15, .01 taper had the highest angle values (P < 0.05). The ProGlider had the lowest angle values in comparison with all the groups (P < 0.05) followed by Mtwo size 10, .04 taper. The Logic size 25, .01 taper had significantly higher angle of rotation values than ProGlider and Mtwo size 10, .04 taper (P < 0.05). CONCLUSION: The Logic size 25, .01 taper instrument made of conventional NiTi alloy had the highest torsional strength of all instruments tested. In addition, the ProGlider instrument manufactured from M-Wire alloy had the lowest angle of rotation to fracture in comparison with the other instruments.

Effect of the combination of several irrigants on dentine surface properties, adsorption of chlorhexidine and adhesion of microorganisms to dentine.

AIM: To investigate the effects of combinations of several irrigants on the roughness and wettability of dentine, adhesion of Enterococcus faecalis and Candida albicans and adsorption of chlorhexidine (CHX) to the dentine. METHODOLOGY: Bovine dentine samples were prepared and their surface roughness standardized. The samples were distributed in groups (n = 10) and subjected to one of the following irrigation protocols: G1 - saline solution; G2 - sodium hypochlorite (NaOCl); G3 - NaOCl + ethylenediaminetetraacetic acid (EDTA); G4 - NaOCl + peracetic acid (PAA); G5 - NaOCl + 1-hydroxyethylidene-1,1-diphosphonic acid (HEDP); G6 - NaOCl + EDTA + CHX; G7 - NaOCl + PAA + CHX; G8 - NaOCl + HEDP + CHX; and G9 - mixture of NaOCl + HEDP. After treatments, roughness and wettability were measured. In order to evaluate the adhesion of microorganisms to dentine, new dentine samples were prepared and after 2 h of contact with the microorganisms, were analysed using a confocal laser scanning microscope and the number of microorganisms adhering to the surfaces were determined. Absorption spectra were collected by attenuated total reflectance of Fourier-transform infrared spectroscopy before and after immersion of other dentine samples in each solution of G6, G7 and G8 and in a solution of 2% CHX at various time intervals. The areas of the band associated with CHX with the peak at 1492 cm-1 were calculated between 1479 and 1500 cm-1 of the spectral range. The data obtained in all experiments were subjected to one-way ANOVA (α < 0.05). The values of the CHX band were also subjected to one-way repeated measures ANOVA (α < 0.05). RESULTS: Saline solution, NaOCl, HEDP and CHX did not alter the roughness of the dentine (P > 0.05), whilst EDTA and PAA did (P < 0.05). Dentine surface wettability increased after the use of all irrigants compared to saline solution (P < 0.05), with HEDP causing the greatest increases (P < 0.05). The adhesion of E. faecalis was favoured on surfaces treated with only saline solution and NaOCl, and on samples that had decalcifying agents as the final irrigant (P < 0.05). The adhesion of C. albicans was highest on surfaces treated with only saline solution and on surfaces that had NaOCl used as the last irrigant (P < 0.05). The use of CHX as a final irrigant reduced the adhesion of both microorganisms. The roughness and wettability did not influence the adhesion of the microorganisms tested. The adsorption of CHX to the dentine was significant after 1 min of immersion of the mineralized samples in the irrigant (P < 0.05), and the use of chelating agents prior to CHX potentiated this adsorption. CONCLUSIONS: The irrigation solutions had a variable effect on the properties of dentine, on the adhesion of E. faecalis and C. albicans and the adsorption of CHX to the dentine surface.

Comparative accuracy of the Clearing Technique, CBCT and Micro-CT methods in studying the mesial root canal configuration of mandibular first molars.

AIMS: To compare the accuracy of the clearing technique and cone beam computed tomography (CBCT) in the assessment of root canal configurations using micro-computed tomography (micro-CT) imaging system as the reference standard. METHODOLOGY: Thirty-two mesial roots of mandibular first molars, selected on the basis of micro-CT scans (voxel size: 19.6 µm) and presenting several canal configurations, were evaluated using 2 CBCT scanners (voxels sizes: 120 µm and 150 µm) followed by the clearing technique. Two examiners analysed the data from each method and classified the anatomical configuration of the mesial canal according to Vertucci's system. Data were compared using Fisher's exact and chi-square tests. Reliability for each assessment was verified by the kappa test, and significance level was set at 5%. RESULTS: Kappa value indicated a high level of agreement between the examiners. Detection of type I configurations was significantly lower in cleared teeth (P < 0.05), whilst type II root canals were detected in all specimens by both tests (P > 0.05). In mesial roots with variable anatomical configurations, CBCT and the clearing method were significantly less accurate than the reference standard (P < 0.05). CONCLUSION: Within the tooth population studied, accuracy of identifying mesial root canal configuration was influenced greatly by the evaluation method and the type of anatomy. Detection of type I configurations in cleared teeth was significantly lower, whilst type II configurations were detected in all specimens by both methods. In mesial roots with variable anatomical configurations, neither CBCT nor clearing methods were accurate for detecting the actual root canal anatomy.

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.

Effect of finishing instrumentation using NiTi hand files on volume, surface area and uninstrumented surfaces in C-shaped root canal systems.

AIM: To assess the effect of 90°-oscillatory instrumentation with hand files on several morphological parameters (volume, surface area and uninstrumented surface) in C-shaped root canals after instrumentation using a single-file reciprocation system (Reciproc; VDW, Munich, Germany) and a Self-Adjusting File System (SAF; ReDent Nova, Ra'anana, Israel). METHODOLOGY: Twenty mandibular second molars with C-shaped canals and C1 canal configurations were divided into two groups (n = 10) and instrumented with Reciproc and SAF instruments. A size 30 NiTi hand K-file attached to a 90°-oscillatory motion handpiece was used as final instrumentation in both groups. The specimens were scanned using micro-computed tomography after all procedures. Volume, surface area increase and uninstrumented root canal surface were analysed using CTAn software (Bruker-microCT, Kontich, Belgium). Also, the uninstrumented root canal surface was calculated for each canal third. All values were compared between groups using the Mann-Whitney test and within groups using the Wilcoxon's signed-rank test. RESULTS: Instrumentation with Reciproc significantly increased canal volume compared with instrumentation with SAF. Additionally, the canal volumes were significantly increased after 90°-oscillatory instrumentation (between and within group comparison; (P < 0.05)). Regarding the increase in surface area after all instrumentation protocols, statistical analysis only revealed significant differences in the within groups comparison (P < 0.05). Reciproc and SAF instrumentation yielded an uninstrumented root canal surface of 28% and 34%, respectively, which was not significantly different (P > 0.05). Final oscillatory instrumentation significantly reduced the uninstrumented root canal surface from 28% to 9% (Reciproc) and from 34% to 15% (SAF; P < 0.05). The apical and middle thirds exhibited larger uninstrumented root canal surfaces after the first instrumentation that was significantly reduced after oscillatory instrumentation (P < 0.05). CONCLUSIONS: The Reciproc and SAF system were associated with similar morphological parameters after instrumentation of mandibular second molars with C-shaped canals except for a higher canal volume increase in the Reciproc group compared to the SAF. Furthermore, the final use of 90°-oscillatory instrumentation using NiTi hand files significantly decreased the uninstrumented canal walls that remained after Reciproc and SAF instrumentation.

Comparison of three retreatment techniques with ultrasonic activation in flattened canals using micro-computed tomography and scanning electron microscopy.

AIM: To use micro-CT to quantitatively evaluate the amount of residual filling material after using several techniques to remove root fillings with and without ultrasonic activation and to analyse the cleanliness of the root canal walls and dentine tubules with scanning electron microscopy (SEM). METHODOLOGY: The root canals of one hundred and eight human mandibular incisors were selected and instrumented with rotary files using the BioRace system up to file size 40, .04 taper. After instrumentation, the teeth were filled using a hybrid technique with gutta-percha and sealer then divided into three groups according to the method used for removing the root filling: G1-Reciproc (using only instrument R50), G2-ProTaper Universal retreatment system and G3-Manual (hand files and Gates-Glidden burs). All groups were divided into two subgroups depending on whether ultrasonic agitation was used with the irrigants. Micro-CT scans were taken before and after removal of the filling material to detect residual material in the canal. After micro-CT analysis, the roots were cut in half, imaged by SEM and scored based on the amount of surface covered by root filling remnants. The data were analysed statistically using a significance level of 5%. RESULTS: All groups had retained material in the root canals after instrumentation. The Reciproc method was associated with less retained material than the ProTaper and Manual methods. Ultrasonic activation significantly reduced the amount of residual root filling in all groups (P < 0.05). CONCLUSION: None of the instrumentation methods were able to completely remove root filling material. Ultrasonic activation improved the removal of root filling material in all groups.

Micro-CT evaluation of C-shaped mandibular first premolars in a Brazilian subpopulation.

AIM: To describe morphometric aspects of the internal anatomy of C-shaped mandibular premolars from a Brazilian subpopulation using micro-CT analysis. METHODOLOGY: First mandibular premolars with radicular grooves (n = 123) were scanned using a micro-computed tomography system. After cross-section analysis, 83 specimens were identified with a C-shaped canal and selected for micro-CT analysis. Number and location of canals according to Vertucci's classification, distances between anatomic landmarks, occurrence of apical deltas, furcation canals, prevalence of C-shaped cross-sections at five levels as well as 2-dimensional analysis (Area, perimeter, roundness, aspect ratio, major and minor diameters) were performed for the more prevalent anatomical features. Data were compared statistically using Kruskal-Wallis tests (α = 0.05). RESULTS: The more prevalent anatomical types according to Vertucci's classification were Type I (13%), III (8%), V (37%) and VII (2%). Mean distances from the furcation to the cemento-enamel junction were in the range of 5.36-5.65 mm. Apical deltas and furcation canals were present in 36 (43%) and 27 (33%) specimens, respectively. C-shaped cross-sections were more prevalent at the middle (56%) and apical middle levels (81%). Overall, significant differences were found in the 2-dimensional analyses between single canals at the apical third and buccal and lingual canals of Vertucci's V classification (P < 0.05). CONCLUSIONS: In this Brazilian subpopulation, C-shaped canal configuration of the root canal system was found in 67% of extracted first mandibular premolars with radicular grooves. Vertucci's types I and V were the most prevalent anatomical variations. C-shaped cross-sections were more prevalent in the middle third, and the presence of apical deltas was the most common feature in the apical third.

Antimicrobial activity of Chlorhexidine, Peracetic acid and Sodium hypochlorite/etidronate irrigant solutions against Enterococcus faecalis biofilms.

AIM: To evaluate the antimicrobial effect of 2.5% sodium hypochlorite alone (NaOCl) and associated with 9% HEBP (NaOCl/HEBP), 2% peracetic acid (PAA) and 2% chlorhexidine (CHX), on the viability of Enterococcus faecalis biofilms attached to dentine. METHODOLOGY: Biofilms of E. faecalis were grown on the surface of dentine blocks for 5 days and then exposed to the irrigating solutions for 3 min. Distilled water was used as the control. The total biovolume and the percentage of dead cells of the infected dentine were measured by means of confocal microscopy and the live/dead technique. Nonparametric tests were used to determine statistical differences (P < 0.05). RESULTS: NaOCl and the NaOCl/HEBP mixture were associated with a significantly greater percentage of dead cells, followed by PAA (P < 0.05). No significant antimicrobial effect of CHX was observed in comparison with the control group. Total biovolume decreased significantly in NaOCl, NaOCl/HEBP and PAA solutions in comparison with the CHX and control groups. CONCLUSIONS: NaOCl alone or associated with HEBP were the most effective irrigant solutions in dissolving and killing E. faecalis biofilms.

Etidronate causes minimal changes in the ability of sodium hypochlorite to dissolve organic matter.

AIM: To evaluate the effect of individual and combined use of sodium hypochlorite (NaOCl), etidronate (HEDP) and ethylenediaminetetraacetic acid (EDTA) in tissue dissolution. METHODOLOGY: Sixty fragments of bovine muscle tissue were prepared and their weights determined on a precision scale. The samples were then distributed in the following groups (n = 10): G1 - saline solution (control); G2 - 17% EDTA; G3 - 18% HEDP; G4 - 2.5% NaOCl; G5 - mixture of 5% NaOCl + 17% EDTA; and G6 - mixture of 5% NaOCl + 18% HEDP. The specimens in each group were immersed in the solutions for 5, 10 and 15 min and reweighted at each time period. Analysis of variance (anova) and Tukey's multiple-comparison tests (α<0.05) were applied to identify the intragroup and intergroup differences. RESULTS: G1, G2, G3 and G5 did not dissolve the organic matter. G4 and G6 significantly reduced the weights of specimens at all periods. Amongst the groups, the difference in ability to dissolve organic matter was greater and significant in the following order G4 = G6 > G5 = G3 = G2 = G1 after 5 min of immersion and G4 > G6 > G5 = G3 = G2 = G1 after 10 and 15 min of immersion. CONCLUSION: The only solution capable of dissolving organic matter was NaOCl. In the mixtures analysed, this ability was arrested by EDTA; however, it was minimally affected by the HEDP, proving that this combination, if used during the biomechanical preparation, is able to dissolve of organic matter.