Effect of pomegranate solution alone or combined with chlorhexidine against oral multispecies biofilm.

AIM: Natural bioactive products have been tested as alternative antimicrobial agents. This study evaluated the effect of Punica granatum extract (PGE) on oral multispecies biofilms. METHODOLOGY: Lyophilized extracts from pomegranate peel were prepared, and the punicalagin content was assessed by ultra-performance liquid chromatography (UPLC). Oral multispecies biofilms from 2 donors were grown on four collagen-coated hydroxyapatite discs. After incubation for 7 days or 3 weeks, the biofilms were exposed to water (control), 2% CHX, 10% PGE, 20% PGE or 30% PGE for 3 min. The proportions of dead bacteria were assessed by the live/dead staining and confocal microscopy. After the analysis, the best PGE concentration (30%) was combined with CHX. The experimental phases were repeated using water, 2% CHX, 30% PGE and 30% PGE + 2% CHX. Five random areas of the biofilm on each disc were scanned, resulting in 20 scanned areas for each group. RESULTS: Regarding the biofilm volume, no differences were found amongst solutions (p = .111). The PGE solution killed bacteria effectively in 1-week, 2-week and 3-week-old-plaque biofilms, ranging from 37 to 55.3%, depending on the PGE concentration. The 30% PGE (a) (p = .0009) had greater antibiofilm effectiveness than 2% CHX (b), which killed bacteria in the 25.2 to 48.7% range. The 10% and 20% PGE had intermediate values (ab), without significant differences from 30% PGE (p = 1.002). Water (c) had the lowest proportion of dead bacteria (p < .00001) in a range of 5 to 6.7% and lower effectiveness in killing bacteria (p < .05). The PGE alone or mixed with 2% CHX had greater anti-biofilm effectiveness than CHX (p < .05). The old plaque biofilms were more resistant than the 7-day-old plaque (p < .05). CONCLUSIONS: The 30% PGE (alone or combined with CHX) exhibited a greater antibiofilm effect on oral multispecies biofilms grown on hydroxyapatite discs than 2% CHX.

Association between genetic polymorphisms in the promoter region of the defensin beta 1 gene and persistent apical periodontitis.

AIM: To evaluate the association between the promoter region of defensin beta 1 (DEFB1) genetic polymorphisms and persistent apical periodontitis (PAP) in Brazilian patients. METHODOLOGY: Seventy-three patients with post-treatment PAP (PAP group) and 89 patients with root filled teeth with healed and healthy periradicular tissues (healed group) were included (all teeth had apical periodontitis lesions at the beginning of the treatment). Patients who had undergone at least 1 year of follow-up after root canal treatment were recalled, and their genomic DNA was extracted from saliva. Two single nucleotide polymorphisms (SNPs) in DEFB1 at the g. -52G>A (rs1799946) and g. -20G>A (rs11362) positions were analysed using real-time polymerase chain reaction. The chi-squared test was performed, and the odds ratios were calculated using Epi Info 3.5.2. Logistic regression analysis in the codominant model, using the time of follow-up as a variable, was used to evaluate the SNP-SNP interaction. All tests were performed with an established alpha of 0.05 (P = 0.05). RESULTS: For the rs11362 polymorphism in the codominant and recessive models, patients who carried two copies of the T allele had a significantly lower risk of developing PAP (P = 0.040 and P = 0.031, respectively). For the rs1799946 polymorphism in DEFB1 in the codominant and recessive models, carrying one copy of the T allele significantly increased the risk of developing PAP (P = 0.007 and P = 0.031, respectively). In the logistic regression, both polymorphisms were associated with PAP as well as the SNP-SNP interaction (P < 0.0001). CONCLUSIONS: Polymorphisms in DEFB1 genes were associated with the development of post-treatment persistent apical periodontitis.

Effect of carbodiimide and chlorhexidine on the bond strength longevity of resin cement to root dentine after radiation therapy.

AIM: To evaluate the effect of carbodiimide (EDC) and chlorhexidine (CHX) on the bond strength (BS) of resin cement to root dentine of teeth submitted to radiotherapy. METHODOLOGY: One hundred and twenty extracted maxillary canines were selected and assigned to 2 groups (n = 60): nonirradiated and irradiated (30 cycles of 2 Gy, total 60 Gy). Roots lengths were standardized, and canals were prepared and filled. Post spaces were then prepared, and the samples were redistributed according to dentine treatment (n = 20): saline solution (SF); CHX 2%; or EDC 0.5M. After drying the post space, fibreglass posts were cemented. Cross-sectioned slices were obtained, and in half of the specimens of each subgroup (n = 10), the analysis was performed immediately; the others (n = 10) were stored for 10 months before analyses. The most cervical slice of each third was subjected to a push-out test and failure pattern analysis (n = 10), and the most apical slice submitted to the analysis of the adhesive interface by SEM (n = 5). The bond strength data were submitted to anova and Tukey tests, the adhesive interface adaptation was submitted to Kruskal-Wallis and Dunn's tests, and the Chi-square test was used to evaluate the type of failure. RESULTS: The irradiated specimens had significantly lower bond strength (13.8 ± 4.3) than the nonirradiated (18.1 ± 3.1; P < 0.001). For the irradiated teeth, the bond strengths were significantly lower in the SF and CHX groups (P < 0.001). Also, the bond strengths reduced significantly after 10 months in the SF and CHX groups (P < 0.001). Cohesive failures occurred in dentine for irradiated specimens. Poorer interface adaptation, dentine fractures and microfractures were observed in irradiated specimens, and better adaptation was observed for specimens after EDC treatment. CONCLUSIONS: Radiotherapy was associated with lower bond strength and worse interface adaptation. Dentine treatment with EDC contributed to adhesive interface longevity during the cementation of glass fibre posts in nonirradiated and irradiated teeth.

Longevity of bond strength of resin cements to root dentine after radiation therapy.

AIM: To evaluate the bond strength and adhesive interface between several resin cements and root dentine immediately and 6 months after radiotherapy. METHODOLOGY: Sixty maxillary canines were selected and randomly assigned to two groups (n = 30): one group was not irradiated and the other one was subjected to a cumulative radiation dose of 60 Gy. The teeth were sectioned to obtain roots 16 mm long and the canals were prepared with the Reciproc system (R50) and filled using a lateral condensation technique with an epoxy resin sealer. Each group was divided into three subgroups (n = 10) according to the resin cement used for fibreglass fibre post cementation: RelyX-U200, Panavia-F2.0 and RelyX ARC. The posts were cemented in accordance with the manufacturer's recommendations. Three 1-mm-thick dentine slices were then obtained from each root third. The first two slices in the crown-apex direction of each third were selected for the push-out test. The failure mode after debonding was determined with a stereo microscope. The third slice from each root third was selected for scanning electron microscopy (SEM) analyses to examine the resin cement-dentine interface with 100, 1000, 2000 and 4000× magnification. Bond strength data were analysed by anova and Tukey's test (α = 0.05). RESULTS: Significantly lower bond strength (P < 0.0001) was obtained after irradiation compared to nonirradiated teeth. RelyX-U200 cemented fibre posts had the higher bond strength (15.17 ± 5.89) compared with RelyX ARC (P < 0.001) and Panavia-F2.0 (P < 0.001). The evaluation after 6 months revealed lower bond strength values compared to the immediate values (P < 0.001) for irradiated and nonirradiated teeth. Cohesive failures occurred in the irradiated dentine. SEM revealed fractures, microfractures and fewer collagen fibres in irradiated root dentine. RelyX-U200 and Panavia-F2.0 were associated with a juxtaposed interface of the cement with the radicular dentine in irradiated and nonirradiated teeth, and for RelyX ARC, hybrid layer formation and tags were observed in both irradiated and nonirradiated teeth. CONCLUSION: Radiation was associated with a decrease in the push-out bond strength and with lower resin cement/root dentine interface adaptation. Self-adhesive resin cement was a better alternative for fibre post cementation in teeth subjected to radiation therapy. The bond strength decreased after 6 months.

Wear profile of canal wall surfaces and bond strength of endodontic sealers after in situ acid challenge.

AIM: To evaluate the wear of root wall surfaces, the bond strength of sealers to dentine and the demineralization around root filling materials after canals were exposed to acid challenge in situ. METHODOLOGY: Eighty-seven roots of mandibular incisors were selected. Thirty-two were used in the laboratory bond strength study (n = 8), and 55 in the in situ study (n = 11). Root canals were prepared biomechanically and then filled with gutta-percha and AH Plus, MTA Fillapex, Sealapex or Endofill. For 14 days, 11 participants used intra-oral devices with five sterilized roots (four experimental and one control - only canal prepared). Drops of sucrose were dripped onto roots allowing the accumulation of biofilm on canal surfaces. Roots were removed, sectioned and analysed for the following: bond strength of filling material using a push-out test and also wear profile and dentine demineralization using confocal microscopy. Bond strength (MPa) was evaluated by two-way anova and Tukey test (α = 0.05), and wear profile was assessed by Kruskal-Wallis and t-tests (α = 0.05). RESULTS: AH Plus had the highest bond strength values. Intermediate results were found in roots with MTA Fillapex and Endofill, whilst Sealapex had inferior results (P < 0.05). No significant differences were found amongst root thirds (P > 0.05). For wear profile, samples were associated with degradation of the filling materials after exposure to the oral environment (P < 0.05). Roots had signs of demineralization around the filling material when Sealapex and Endofill were used. CONCLUSIONS: Sealers were not able to prevent degradation of the adhesive interface and dentine. AH Plus and MTA Fillapex had superior bond strength to dentine and less intense demineralization around the root filling.

Effect of ultrasonic and sonic activation of root canal sealers on the push-out bond strength and interfacial adaptation to root canal dentine.

AIM: To evaluate the effect of ultrasonic and sonic activation of two root canal sealers on interfacial adaptation and push-out bond strength (BS) to root canal dentine. METHODOLOGY: The root canals of seventy-eight roots of maxillary canines were prepared with NiTi rotary instruments under irrigation with 2.5% NaOCl and 17% EDTA. Canals were filled by lateral condensation of gutta-percha and AH Plus (n = 39) or MTA Fillapex (n = 39). Three subgroups (n = 13) were formed according to sealer activation: no activation (NA), sonic activation (SA, 20 s) and ultrasonic activation (US, 20 s). In three specimens of each subgroup, 0.1% rhodamine B was added to the sealer. Three 1-mm-thick slices were obtained from each root third. The first and second were used for push-out BS test and the third for a qualitative analysis of interfacial adaptation (gaps) and voids by confocal laser scanning microscopy (CLSM). Intratubular penetration of rhodamine B-labelled sealer was also assessed by CLSM. Data were analysed by two-way anova and Tukey's test (α = 0.05). RESULTS: AH Plus (2.58 ± 1.21) had higher push-out BS than MTA Fillapex (1.45 ± 0.71) (P = 0.000). UA (2.64 ± 1.44) had higher BS (P = 0.000) than NA (1.58 ± 0.73) and SA (1.83 ± 0.85), which did not differ significantly from each other (P = 0.3303). The coronal third had higher BS (2.48 ± 1.49) than the middle (2.08 ± 0.94) (P = 0.0003) and apical (1.48 ± 0.58) (P = 0.00) thirds. For both sealers, UA was associated with a more homogeneous filling material with better interfacial adaptation and greater quantity, density and length of tags in dentinal tubules. When UA was used, the tags had similar quantity, density and length to those formed without activation, but interfacial gaps and voids were observed. CONCLUSION: Ultrasonic activation was associated with higher BS values, deeper intratubular penetration and greater interfacial adaptation to root dentine than sonic activation and no activation techniques, AH Plus had higher BS values than MTA Fillapex irrespective of the type of activation.

Evaluation of bond strength in single-cone fillings of canals with different cross-sections.

AIM: To evaluate the effect of root canal cross-sectional shape on single-cone root filling bond strength, as well as to determine the percentage of gutta-percha-filled areas (PGFA) and sealer-filled areas (PSFA), establishing a relationship between these variables. METHODOLOGY: Distal roots of mandibular molars were selected using microcomputed tomography imaging and allocated into three groups (n = 10) according to canal shape: round, oval and long oval. The canals were prepared with an R40 reciprocating instrument and filled with matching single-cone gutta-percha and AH Plus sealer. Two 1-mm-thick dentine slices were obtained from each third of each root. PGFA and PSFA were calculated in digital images (x25 magnification) of each slice. Next, the slices were subjected to a push-out test and the failure modes (adhesive, cohesive or mixed) were assessed. Data were analysed using parametric tests (P < 0.05). RESULTS: In the coronal (2.17 ± 0.56MPa) and middle thirds (1.78 ± 0.45MPa), the round canals were associated with higher bond strength values (P < 0.01), with no difference between the groups for the apical third (P > 0.05). Adhesive and mixed failures predominated in round canals, whilst cohesive failures were more frequent in oval and long oval canals. Round canals had significantly higher PGFA and lower PSFA than all other groups (P = 0.000). The PGFA and PSFA had a positive (r = 0.521, P = 0.000) and a negative (r = -0.523, P = 0.000) correlation with bond strength, respectively. CONCLUSION: Bond strength values of gutta-percha and sealer were affected by canal shape. Higher percentage of gutta-percha-filled area resulted in higher bond strength to dentine.

Ex vivo evaluation of four final irrigation protocols on the removal of hard-tissue debris from the mesial root canal system of mandibular first molars.

AIM: To evaluate the efficacy of four final irrigation protocols on the reduction of hard-tissue debris accumulated within the mesial root canal system of mandibular first molars using micro-CT analysis. METHODOLOGY: Forty mesial roots of mandibular molars with a single and continuous isthmus connecting the mesiobuccal and mesiolingual canals (Vertucci's Type I configuration) were selected and scanned at a resolution of 8.6 µm. Canals were enlarged sequentially using WaveOne Small and Primary instruments activated in reciprocating motion without intracanal irrigation to allow debris to accumulate within the mesial root canal system. Then, specimens were anatomically matched and distributed into four groups (n = 10), according to the final irrigation protocol: apical positive pressure (APP), passive ultrasonic irrigation (PUI), Self-adjusting File (SAF) and XP-endo Finisher (XPF). The final irrigation procedures were performed over 2 min using a total of 5.5 mL of 2.5% NaOCl per canal. Reconstructed data sets were coregistered, and the mean percentage reduction of accumulated hard-tissue debris after the final irrigation procedures was compared statistically between groups using the anovapost hoc Tukey test with a significance level set at 5%. RESULTS: Reduction of accumulated hard-tissue debris was observed in all groups after the final irrigation protocol. Overall, PUI and XPF groups had higher mean percentage reductions of accumulated hard-tissue debris (94.1% and 89.7%, respectively) than APP and SAF groups (45.7% and 41.3%, respectively) (P < 0.05). No significant differences were found when comparing the results of PUI and XPF groups (P > 0.05) or APP and SAF groups (P > 0.05). CONCLUSIONS: The PUI technique and XP-endo Finisher instrument were associated with significantly lower levels of AHTD compared with conventional irrigation and the modified SAF system protocol in mesial root canals of mandibular molars.

Effects of different lasers on organic/inorganic ratio of radicular dentin.

The aim of this study was to evaluate the chemical stability of endodontic-treated root dentin after different laser irradiations through Raman spectroscopy. Fifty maxillary canines were selected and prepared with K3 system. Roots were randomly distributed into five groups (n = 10) according to the surface treatment: GI (water), GII (NaOCl + EDTA), GIII (NaOCl + EDTA + 980 nm Diode laser), GIV (NaOCl + EDTA+ 1064 nm Nd: YAG laser) and GV (NaOCl + EDTA+ 2780 nm Er,Cr: YSGG laser). Lasers were applied for 20 s. Samples were bisected, and the organic and inorganic content of dentin was analyzed by Raman spectroscopy. Data were submitted to ANOVA and Tukey tests (p < 0.05). None of the surface treatments alter the inorganic content (cts) (p = 0.183). Roots irradiated with Er,Cr: YSGG laser had a reduced collagen content (GV-290.7 ± 41.7) compared with the water-treated roots (GI-328.3 ± 63.5) and those treated with NaOCl + EDTA (GII-333.9 ± 55.8). Roots irradiated with Er,Cr: YSGG laser also showed a higher inorganic/organic ratio (GV-9.5 ± 1.1) than roots treated with water (GI-7.7 ± 1.5), NaOCl + EDTA (GII-8.0 ± 1.4) and diode laser (GIII-8.2 ± 1.6). Both organic and inorganic contents increased from cervical to apical thirds in all groups. None of the surface treatments were able to promote changes in the inorganic content of the root dentin; treatment with NaOCl + EDTA combined with Er,Cr: YSGG altered collagen.

Micro-CT evaluation of the efficacy of hard-tissue removal from the root canal and isthmus area by positive and negative pressure irrigation systems.

AIM: To evaluate the removal of accumulated hard-tissue debris (AHTD) from the root canal system of mandibular molars by positive and negative pressure irrigation systems, using micro-CT imaging analysis. METHODOLOGY: Mandibular molars with a single canal in the distal root and 2 canals connected by an isthmus in the mesial root were matched based on similar morphological dimensions using micro-CT evaluation and assigned to 2 experimental groups (n = 20 mesial and 10 distal canals), according to the irrigation protocol: apical positive (conventional irrigation) or negative (EndoVac system) pressure. Changes in root canal volume and surface area as well as percentage of uninstrumented canal wall surface and accumulated hard-tissue debris (AHTD) after canal preparation were compared statistically using the independent sample t-test and Mann-Whitney U-test, with the significance level set at 5%. RESULTS: Volume, surface area and percentage of static voxels in either mesial or distal root canal systems were not significantly different between groups before or after root canal preparation (P > 0.05). After preparation, AHTD was not observed in the distal canal of both groups. However, in the mesial root canal system, the conventional irrigation group was associated with a significantly higher median percentage of AHTD (11.48%; IQR: 5.9-22.6; range: 1.86-41.98) than the EndoVac group (3.40%; IQR: 1.5-7.3; range: 0.82-12.84) (P < 0.05). CONCLUSIONS: Neither irrigation protocol succeeded in rendering the mesial canal system free of AHTD; however, apical negative pressure irrigation resulted in lower levels of AHTD than conventional irrigation.

Influence of therapeutic cancer radiation on the bond strength of an epoxy- or an MTA-based sealer to root dentine.

AIM: To evaluate the influence of radiation on root canal sealer push-out bond strength to dentine and sealer/dentine interface in teeth filled with AH Plus (Dentsply Ind. Com. Ltda, Petrópolis, RJ, Brazil) and MTA Fillapex (Angelus Ind. Prod. Odontológicos S/A, Londrina, PR, Brazil). METHODOLOGY: Thirty-two maxillary canines were selected and randomly assigned to 2 groups (n = 16): one group was not irradiated, and the other was subjected to a cumulative radiation dose of 60 Gy. Root canals were prepared with the Reciproc system (VDW GmbH, Munich, Germany), and each group was divided into 2 subgroups (n = 8) according to the sealer - AH Plus or MTA Fillapex - using the single-cone filling technique. Then, 1-mm-thick dentine slices were obtained from each root third for the push-out test to evaluate sealer bond strength to dentine and for scanning electron microscopy (SEM) to examine the sealer/dentine interface. Failure mode after debonding was determined with a stereomicroscope at ×25 magnification. Bond strength data were analysed by two-way anova with a split-plot design and post hoc Tukey's test (α = 0.05). RESULTS: Significantly lower bond strength (P < 0.0001) was obtained after irradiation (0.71 ± 0.20 versus 0.97 ± 0.29 MPa) and in specimens filled with MTA Fillapex (0.70 ± 0.18 MPa) compared with AH Plus (1.00 ± 0.27 MPa). Percentage of adhesive failures increased after radiation in all root thirds in the teeth filled with AH Plus. SEM revealed more gap-containing regions and fewer tags at the sealer/dentine interface in irradiated specimens, with more tag formation and fewer gaps with AH Plus sealer. CONCLUSIONS: Radiation was associated with a decrease in the push-out bond strength of sealers to intraradicular dentine and formation of more gaps and fewer tags at the sealer/dentine interface regardless of the sealer.

Use of a dissolved oxygen microsensor for assessing the viability and thickness of microbial biofilm on root surfaces.

AIM: To evaluate the use of a dissolved oxygen microsensor (DOMS) for assessing the viability and thickness of microbial biofilms on the apical external surface of contaminated human tooth roots. METHODOLOGY: Apical biofilm formation was evaluated in 15 roots contaminated in vitro with a polymicrobial mixture of Staphylococcus aureus, Enterococcus faecalis, Pseudomonas aeruginosa and Candida albicans for 7, 21 and 60 days and in three freshly extracted roots with associated radiographically visible periapical lesions. In each root, the thickness and viability (measured by the amount of dissolved oxygen) of biofilm formed on the apical 2 mm were examined with the DOMS. Scanning electron microscopy (SEM) was used as an auxiliary analysis to confirm the existence of the biofilms detected by the DOMS. RESULTS: The DOMS detected dissolved oxygen on the biofilms formed on the three residual roots up to thickness of 375 µm, 480 µm and 1650 µm. In the 15 roots contaminated in vitro, the DOMS detected dissolved oxygen in six specimens up to thicknesses from 75 to 250 µm, and the intensity of the metabolic activity (biofilm thickness) was directly proportional to the contamination time. SEM confirmed the presence of biofilm in all roots. CONCLUSION: The dissolved oxygen microsensor allowed the measurement of the amount of dissolved oxygen in the biofilm, which is indicative of the intensity of the microbial metabolic activity (viability), correlating the results with biofilm thickness. The DOMS was effective in freshly extracted roots, but had limitations in roots contaminated in vitro after short periods (7 and 21 days) of contamination.

Shaping ability of single-file reciprocating and heat-treated multifile rotary systems: a micro-CT study.

AIM: To investigate changes in three-dimensional geometry, in various cross-sectional morphological parameters and in the centring ability of root canals prepared with different preparation systems using microcomputed tomographic imaging technology. METHODOLOGY: Sixty-four mesial canals of mandibular molars were matched based on similar morphological dimensions using micro-CT evaluation and assigned to four experimental groups (n = 16), according to the canal preparation technique: Reciproc, WaveOne, Twisted File and HyFlex CM systems. Changes in several 2D (area, perimeter, form factor, roundness, minor and major diameter) and 3D [volume, surface area, structure model index (SMI)] morphological parameters, as well as canal transportation, were compared with preoperative values using Kruskal-Wallis and anovapost hoc Tukey's tests with the significance level set at 5%. RESULTS: Preparation significantly increased all tested parameters in the experimental groups. No significant differences were observed between groups regarding changes in volume, surface area, SMI, form factor and roundness of the root canal after preparation (P > 0.05). In the apical third, the Reciproc group had significantly greater changes in canal area, perimeter, major and minor diameters than the other groups (P < 0.05). Overall, the Twisted File and HyFlex CM systems were associated with significantly less transportation than the reciprocating instruments, Reciproc and WaveOne (P < 0.05). CONCLUSIONS: Shaping procedures led to the enlargement of the root canal space with no evidence of significant preparation errors. Changes in 3D parameters were not different between groups whilst, in the apical third, Reciproc was associated with significantly greater changes in several 2D parameters compared to the other groups. Twisted File and HyFlex CM systems were able to maintain the original canal anatomy with less canal transportation than Reciproc and WaveOne; however, these differences are unlikely to be of clinical significance.

Root canal morphology of primary molars: a micro-computed tomography study.

AIM: This was to investigate the root canal morphology of primary molar teeth using micro-computed tomography. METHODS: Primary maxillary (n = 20) and mandibular (n = 20) molars were scanned at a resolution of 16.7 µm and analysed regarding the number, location, volume, area, structured model index (SMI), area, roundness, diameters, and length of canals, as well as the thickness of dentine in the apical third. Data were statistically compared by using paired-sample t test, independent sample t test, and one-way analysis of variance with significance level set as 5%. RESULTS: Overall, no statistical differences were found between the canals with respect to length, SMI, dentine thickness, area, roundness, and diameter (p > 0.05). A double canal system was observed in the mesial and mesio-buccal roots of the mandibular and maxillary molars, respectively. The thickness in the internal aspect of the roots was lower than in the external aspect. Cross-sectional evaluation of the roots in the apical third showed flat-shaped canals in the mandibular molars and ribbon- and oval-shaped canals in the maxillary molars. CONCLUSIONS: External and internal anatomy of the primary first molars closely resemble the primary second molars. The reported data may help clinicians to obtain a thorough understanding of the morphological variations of root canals in primary molars to overcome problems related to shaping and cleaning procedures, allowing appropriate management strategies for root canal treatment.

Fracture resistance and stress distribution of simulated immature teeth after apexification with mineral trioxide aggregate.

AIM: To evaluate the effect of adhesive restorations on fracture resistance and stress distribution in teeth with simulated immature apices and apical plugs of mineral trioxide aggregate (MTA). METHODOLOGY: Sixty bovine incisors were sectioned 8 mm above and 12 mm below the cemento-enamel junction (CEJ). The root canal was enlarged using a diamond bur, resulting in remaining root canal walls with 0.1-0.2 mm of thickness. A 5-mm apical plug of MTA was placed and the teeth were restored according to the following groups: GP--the root canal was filled with gutta-percha and endodontic sealer; CR--the root canal was filled with light-cured composite resin inserted incrementally; FP--a fibre post was cemented into the root canal; and RFP--the fibre post was relined with composite resin prior to the cementation into the root canal. A load was applied on the crown of all teeth at 135° to their long axis until fracture. Data was analysed by one-way anova and SNK tests (α = 0.05), whilst the fracture pattern was evaluated according to the position of the fracture. Stress distributions in the restored teeth were verified by finite element analysis. RESULTS: Teeth restored with fibre posts and relined fibre posts were associated with the highest fracture resistance, whilst the GP group had the lowest values. GP and RC groups had similar fracture resistance values (P = 0.109). All fractures types involved the cervical and middle thirds of roots. The GP model had high levels of stress concentration in the cervical and middle thirds of roots. No difference was found amongst the stress concentration in the RC, FP and RFP models. CONCLUSION: Restorative protocols alter the fracture resistance and stress distribution of immature teeth after placement of MTA apical plugs.

Confocal microscopy assessment of filling material remaining on root canal walls after retreatment.

AIM: The percentage of Endofill remaining on canal walls after retreatment with different techniques was evaluated using confocal microscopy and qualitative analysis of the interface between the filling material/dentine. METHODOLOGY: Sixty-four root canals of incisors were prepared with ProTaper, filled with gutta-percha and Endofill mixed with 0.1% rhodamine B. The roots were thermocycled and distributed into groups according to the method of evaluation: GI - direct viewing (DV) and GII - operating microscope (OM) and according to the removal technique: A) ProTaper retreatment (PR), B) PR/xylol, C) ultrasound and D) ultrasound/xylol. The root canals were then refilled with gutta-percha and AH Plus with 0.1% fluorescein and sectioned at 2, 4 and 6 mm from the apex. The percentage of remaining Endofill was analysed by confocal microscopy. Additionally, 16 roots were prepared with a ProTaper F5 instrument and were filled with Endofill + 0.1% rhodamine B/gutta-percha (negative control group) (n = 8), and the positive control group (n = 8) were filled with AH Plus with 0.1% fluorescein/gutta-percha. RESULTS: Three-way anova demonstrated differences in the method of evaluation, removal techniques and their interaction (P < 0.05). OM (26.15 ± 12.16%) had a smaller percentage of remaining sealer than DV (32.77 ± 14.47%). The Tukey's test revealed that ultrasound/xylol (15.77 ± 7.15%) led to lower percentages of remaining sealer, significantly different from the PR group (35.25 ± 13.63%), PR/xylol (33.03 ± 11.64%) and ultrasound (33.79 ± 11.71%), which were similar (P > 0.05). Qualitative analysis detected that ultrasound had lower remaining Endofill than PR, particularly when combined with xylol. Operating microscope resulted in lower residual sealer, regardless of the removal technique. CONCLUSIONS: None of the protocols was associated with complete removal of the filling material; however, the use of ultrasound/xylol under an OM provided better results.

Impact of remaining zinc oxide-eugenol-based sealer on the bond strength of a resinous sealer to dentine after root canal retreatment.

AIM: To evaluate the bond strength of a resin-based sealer (AH Plus) to root canal dentine after the removal of a zinc oxide-eugenol-based sealer (Endofill), using different retreatment techniques. METHODOLOGY: The root canals of sixty-four maxillary incisors were prepared with ProTaper and filled with Endofill/gutta-percha. After thermocycling, the roots were randomly assigned according to the retreatment technique (n = 16): (A) ProTaper, (B) ProTaper/xylol, (C) ultrasound and (D) ultrasound/xylol and according to whether an operating microscope (OM) was used or not (n = 8): GI- direct vision (DV) and GII- indirect through the OM. Eight additional roots were instrumented and filled with AH Plus/gutta-percha (control). Roots were sectioned to obtain 1-mm-thick slices. Slices were used in the push-out test and in confocal microscopy. Data were submitted to anova and Tukey test (α = 0.05). RESULTS: The specimens filled under the OM (2.50 ± 0.90) had higher bond strength values of sealer to root canal walls (P < 0.05). The control group had superior mean adhesion values (4.75 ± 1.47) that were significantly different from the others (P < 0.05). Ultrasound/xylol (2.53 ± 1.00) was superior to ultrasound (2.23 ± 0.72) (P < 0.05), ProTaper (2.20 ± 0.69), ProTaper/Xylol (2.12 ± 0.65), which were similar (P > 0.05). Bond strength was different in all thirds (P < 0.05) and decreased in the following sequence: cervical (3.19 ± 1.18), middle (2.58 ± 1.21) and apical (1.87 ± 0.68). Confocal analysis revealed residues of Endofill in dentinal tubules when the removal protocol was carried out with ProTaper, but not from ultrasound combined with xylol. CONCLUSIONS: The OM was associated with higher bond strength values for filling material to root canals. The zinc oxide-eugenol-based sealer negatively affected the bond strength of AH Plus to root canal walls, regardless of the retreatment technique.

Ex vivo evaluation of artifacts mimicking fracture lines on cone-beam computed tomography produced by different root canal sealers.

AIM: To evaluate streaking artifacts produced by several root canal sealers on CBCT images with variations in voxel resolution. METHODOLOGY: Single-rooted premolars were selected and root filled using gutta-percha cones and one of the following sealers: Endofill, Sealer 26, Fillapex or AH Plus. Root canals filled only with gutta-percha were used as control. Scans of teeth using a small-volume CBCT were performed, and the images acquired were evaluated by two calibrated observers (k > 0.75). Artifacts were identified as streaks and dark bands on root dentine in axial slices with 2 voxel sizes (0.076 and 0.2 mm). Fisher's exact test (P < 0.05) was used to compare the presence/absence ratio of artifacts considering the factors 'sealer' and 'voxel size'. RESULTS: For the 0.20 mm voxel size, Sealer 26 had significantly more artifacts than Fillapex (P = 0.006) or AH Plus (P = 0.017). No differences existed between the sealers when the 0.07 mm voxel size was used. Sealer 26 (P = 0.042) and Endofill (P = 0.035) had significantly more artifacts in the 0.20 mm voxel size than with the 0.076 mm voxel size, whilst no difference was noted for other filling conditions. CONCLUSIONS: The material used for root filling and the voxel resolution can influence the presence of streaking artifacts observed in CBCT images.

Enamel pearls in permanent dentition: case report and micro-CT evaluation.

OBJECTIVES: To investigate the frequency, position, number and morphology of enamel pearls (EPs) using micro-CT (µCT) and to report a case of an EP mimicking an endodontic-periodontic lesion. METHODS: Cone beam CT (CBCT) was performed in a patient to evaluate a radio-opaque nodule observed on the left maxillary first molar during the radiographic examination. Additionally, 23 EPs were evaluated regarding frequency, position, number and morphology by means of µCT. The results were statistically compared using the Student's t-test for independent samples. RESULTS: 1 pearl was presented in 13 specimens, while 5 specimens presented 2 pearls. The most frequent location of the EPs was the furcation between the disto-buccal and the palatal roots of the maxillary molars. Overall, the mean major diameter, volume and surface area were 1.98 ± 0.85 mm, 1.76 ± 1.36 mm³ and 11.40 ± 7.59 mm², respectively, with no statistical difference between maxillary second and third molars (p > 0.05). In the case report, CBCT revealed an EP between the disto-buccal and the palatal roots of the maxillary first left molar associated with advanced localized periodontitis. The tooth was referred for extraction. CONCLUSIONS: EPs, located generally in the furcation area, were observed in 0.74% of the sample. The majority was an enamel-dentin pearl type and no difference was found in maxillary second and third molars regarding diameter, volume and surface area of the pearls. In this report, the EP mimicked an endodontic-periodontic lesion and was a secondary aetiological factor in the periodontal breakdown.

Microcomputed tomography analysis of the root canal morphology of single-rooted mandibular canines.

AIM: To investigate the anatomy of single-rooted mandibular canine teeth using microcomputed tomography (µCT). METHODOLOGY: One hundred straight single-rooted human mandibular canines were selected from a pool of extracted teeth and evaluated using µCT. The anatomy of each tooth (length of the roots, presence of accessory canals and apical deltas, position and major diameter of the apical foramen and distance between anatomical landmarks) as well as the two- and three-dimensional morphological aspects of the canal (area, perimeter, form factor, roundness, major and minor diameter, volume, surface area and structure model index) were evaluated. The results of the morphological analysis in each canal third were compared statistically using Friedman's test (α = 0.05). RESULTS: The length of the roots ranged from 12.53 to 18.08 mm. Thirty-one specimens had no accessory canals. The location of the apical foramen varied considerably. The mean distance from the root apex to the major apical foramen was 0.27 ± 0.25 mm, and the major diameter of the major apical foramen ranged from 0.16 to 0.72 mm. Mean major and minor diameters of the canal 1 mm short of the foramen were 0.43 and 0.31 mm, respectively. Overall, the mean area, perimeter, form factor, roundness, major and minor diameters, volume, surface area and structure model index (SMI) were 0.85 ± 0.31 mm(2) , 3.69 ± 0.88 mm, 0.70 ± 0.09, 0.59 ± 0.11, 1.36 ± 0.36 mm and 0.72 ± 0.14 mm, 13.33 ± 4.98 mm(3) , 63.5 ± 16.4 mm(2) and 3.35 ± 0.64, respectively, with significant statistical difference between thirds (P < 0.05). CONCLUSIONS: The anatomy and morphology of the root canal of single-rooted canines varied widely in different levels of the root.