Effect of ELVAX polymer subgingival implants with echistatin on extracted and reimplanted rats' teeth.

To investigate the effect of ELVAX polymer subgingival implants incorporated with echistatin peptide on incisor reimplanted tooth in rats. Forty-two male Wistars rats were divided into two groups: echistatin-treated rats (E) and control rats (C). The animals had their right maxillary incisors extracted and treated according to the International Association of Dental Traumatology replantation protocol. The extra-alveolar dry period was 30 and 60 min, and the post-surgical experimental periods were 15, 60, and 90 days. The samples were stained with H&E and analyzed for the presence of an inflammatory response, incidence of resorptions, and dental ankylosis. Results were statistically analyzed (p < 0.05). The presence of inflammatory resorption was significantly higher in group C at 30 and 60 min extra-alveolar time, in the 15-day postoperative period as compared with the E group (p < 0.05). Dental ankylosis was significantly more prevalent in group E in 30 min extra-alveolar time and 15 days postoperative period (p < 0.05). However, in 60 min extra-alveolar time and 60 days postoperative period, dental ankylosis was more prevalent in C group (p < 0.05). The use of ELVAX subgingival implants with echistatin demonstrated therapeutic potential in preventing the experimental resorption process after replantation of maxillary incisors in rats.

The antimicrobial effect of new and conventional endodontic irrigants on intra-orally infected dentin.

OBJECTIVES: To evaluate if the incorporation of antimicrobial compounds to chelating agents or the use of chelating agents with antimicrobial activity as 7% maleic acid and peracetic acid show similar disinfection ability in comparison to conventional irrigants as sodium hypochlorite or iodine potassium iodide against biofilms developed on dentin. MATERIALS AND METHODS: The total bio-volume of live cells, the ratio of live cells and the substratum coverage of dentin infected intra-orally and treated with the irrigant solutions: MTAD, Qmix, Smear Clear, 7% maleic acid, 2% iodine potassium iodide, 4% peracetic acid, 2.5% and 5.25% sodium hypochlorite was measured by using confocal microscopy and the live/dead technique. Five samples were used for each irrigant solution. RESULTS: Several endodontic irrigants containing antimicrobials as clorhexidine (Qmix), cetrimide (Smear Clear), maleic acid, iodine compounds or antibiotics (MTAD) lacked an effective antibiofilm activity when the dentin was infected intra-orally. The irrigant solutions 4% peracetic acid and 2.5-5.25% sodium hypochlorite decrease significantly the number of live bacteria in biofilms, providing also cleaner dentin surfaces (p < 0.05). CONCLUSIONS: Several chelating agents containing antimicrobials could not remove nor kill significantly biofilms developed on intra-orally infected dentin, with the exception of sodium hypochlorite and 4% peracetic acid. Dissolution ability is mandatory for an appropriate eradication of biofilms attached to dentin.

Effect of sputter-coating on cracking of root-end surfaces after ultrasonic retrograde preparation--a SEM study of resected root apices and their respective impressions.

The aim of this study was to evaluate in vitro the effect of sputter-coating for SEM analysis on the formation of cracks on root-end surfaces after retrograde cavity preparation with ultrasonic tips. Root-end cavities were prepared with either Satelec S12/90 degrees D diamond-coated or S12/90 degrees non-coated stainless steel retrotips. Impressions were taken before and after retrograde cavity preparation. The resected root apices and their respective impressions were examined using a scanning electron microscope, and the presence, extension and numbers of cracks were recorded after each procedure. The number of cracks observed directly on the ultrasonically prepared root-end surfaces was larger than that observed in their respective impressions taken after root-end cavity preparation, which suggests that cracking was mostly produced by the sputter-coating process required for SEM analysis. Impressions of the root-end cavities prepared with non-coated ultrasonic stainless steel retrotips showed a greater incidence of cracks (3/10 impressions) than those that replicated cavities prepared with diamond-coated retrotips (1/10 impressions). No statistical difference was found between the diamond and stainless steel retrotips.

Analysis of the physicochemical properties, cytotoxicity and volumetric changes of AH Plus, MTA Fillapex and TotalFill BC Sealer.

BACKGROUND: To evaluate the physicochemical properties and cytotoxicity of AH Plus, MTA Fillapex and TotalFill BC Sealer. Volumetric changes were also evaluating using micro-computed tomography (micro-CT). MATERIAL AND METHODS: Radiopacity and flow were evaluated in accordance with the ISO 6876, while setting time was evaluated in accordance with the ASTM- C266-08 specifications. The release of Ca2+ ions and pH were measured with spectrophotometer and pH meter, respectively, after different time intervals (1h, 3h, 24h, 72h, 168h, and 360h). Cytotoxicity was evaluated by MTT reduction assay to check 3T3 cells viability at 24, 48 and 72 hours. Volumetric change was evaluated by micro-CT, by using 30 acrylic teeth, filled with gutta-percha cones and the tested root canal sealer. The samples were evaluated after 168h, 360h and 720h of immersion in distilled water. Data were statistically analyzed by one-way ANOVA and Tukey test or by Kruskal-Wallis and Dunn tests (P<0.05). RESULTS: MTA Fillapex and TotalFill BC Sealer showed lower radiopacity than AH Plus (P<0.05). The MTA Fillapex showed the highest flow, while AH Plus showed the lowest flow (P<0.05). The initial and final setting time of AH Plus were lower than MTA Fillapex and TotalFill BC Sealer (P<0.05). In general, TotalFill BC Sealer presented higher Ca2+ ion release and pH than the other tested sealers. TotalFill BC Sealer also showed overall lower cytotoxicity when compared to the other sealers. Volumetric change of AH Plus and TotalFill BC Sealer was lower than MTA Fillapex (P<0.05). CONCLUSIONS: AH Plus, MTA Fillapex and TotalFill BC Sealer showed slight differences in the physicochemical properties and cytotoxicity, but all suitable for an endodontic sealer. However, AH Plus and TotalFill BC Sealer showed low volumetric changes when compared to MTA Fillapex. Key words:Calcium silicate, cytotoxicity, physicochemical properties, micro computed tomography.

CAMPANELLIProgrammed Death 1 (PD-1) and PD-1 Ligand (PD-L1) Expression in Chronic Apical Periodontitis.

OBJECTIVE: This study aimed to examine programmed death protein 1 (PD-1) and programmed death lig-and 1 (PD-L1) expression on leukocytes from chronic apical periodontitis, and to determine the levels of cytokines in the apical periodontitis lesions. METHODS: Leukocytes from healthy gingival tissue (n=16) and chronic apical periodontitis (n=10) were eval-uated using flow cytometry. The PD-1 and PDL-1 expressions were evaluated using flow cytometry. The cy-tokine levels were evaluated by enzyme-linked immunosorbent assay. Data were analyzed using one-way ANOVA. The statistical significance level was set at P<0.05. RESULTS: Results showed that the apical periodontitis lesions are more infiltrated by PD-1+ and PDL1+ lym-phocytes than the control samples. In addition, the PDL-1 expression was detected on macrophages in the apical periodontitis lesions, and was significantly higher compared to leukocytes from healthy gingival tis-sue. The IFN-γ, TGF-ß, IL-10, and TNF-α levels were significantly higher in the apical periodontitis lesions com-pared to control samples. CONCLUSION: The PD-1, PD-L1, and CTLA-4 molecules are evident in apical periodontitis, and can be an impor-tant immune checkpoint in chronic periapical periodontitis.

Evaluation of the temperature of different refrigerant sprays used as a pulpal test.

The temperature of different refrigerant sprays (Endo-Ice, Endo-Frost, Coolermatic and Sprayon Contact and Tuner Cleaner) used as pulpal tests were evaluated in vitro. A thermocouple placed inside the pulp chamber of a maxillary central incisor was used to register the temperature changes when the refrigerant sprays were applied with a cotton swab, for 10 s. Results indicate that Endo-Ice and Endo-Frost presented the lowest temperatures among the refrigerant sprays tested. Temperatures measured inside the pulp chamber, however, were statistically similar in all groups.

Influence of embedding media on the assessment of electronic apex locators.

Third-generation electronic apex locators for root canal length determination are very reliable and are not subject to interference from the contents of the canals. This study compared the effectiveness of different embedding media for in vitro assessment of electronic apex locators. The tooth lengths of 30 extracted human mandibular central incisors were measured by introducing a size 15 K-file fitted with a silicone stop into the canal until its tip appeared through the apical foramen; the distance between the tip and stop was measured. The teeth were placed in cylindrical polyethylene tubes filled with different embedding media (1% agar, gelatin, alginate, saline, and flower sponge soaked in saline), and electronic reading was performed with the Root ZX device. Data were statistically assessed by the analysis of variance (ANOVA) and Tukey tests at a significance level of 5%. Despite the lack of statistically significant difference among the media, the flower sponge was the only medium in which the file surpassed the apex in some measurements.

Evaluation of apical cavity preparation with a new type of ultrasonic diamond tip.

This study evaluated the time, occurrence of fracture, and quality of apical cavity preparation with three different ultrasonic diamond tips: Satelec, Trinity, and a new type, CVD (chemical vapor deposition), using scanning electron microscopy (SEM) analysis. Thirty human single-rooted premolars were selected, submitted to apicectomy, and prepared with ultrasonic tips; impressions were then obtained. The presence of fractures was evaluated on the impressions, and the quality of preparation was evaluated by SEM analysis of teeth and scoring by two examiners. The group prepared with the CVD tips exhibited the shorter preparation time and did not present fractures. There was no statistically significant difference in the quality of preparation for the three tips. The three brands of ultrasonic tips produced adequate grinding without altering the morphology of the apical foramen.

Endodontic treatment of dens in dente associated with a chronic periapical lesion using an apical plug of mineral trioxide aggregate.

Dens in dente is a developmental anomaly that predisposes the tooth to develop caries and pulp periapical pathologies. A chronic lesion can develop without any clinical symptoms of the pathology. The purpose of this case report is to describe the endodontic treatment of dens in dente associated with a periapical lesion using a mineral trioxide aggregate apical plug.

Comparison of two methods of irrigant agitation in the removal of residual filling material in retreatment.

The aim of this study was to compare the efficacy of passive ultrasonic irrigation and EasyClean for removing residual filling material in retreatment. Twenty-two maxillary lateral incisors with apical curvature were instrumented with ProTaper files and filled with Endofill using the lateral compactation technique. Removal of filling material was performed with Reciproc, Mtwo and ProDesign Logic 50/.01 files. The teeth were inserted in a silicone mould, which was placed in a metal muffle, and split to visualize the residual filling material. The samples were divided into two groups (n = 11) according to the irrigation protocol: Passive ultrasonic irrigation (PUI group) with 3 activations of 20 seconds and EasyClean (Easy Equipamentos Odontológicos, Belo Horizonte, Brazil) (EC group) used in continuous rotation with 3 activations of 20 seconds, both using NaOCl and EDTA. Environmental scanning electron microscopic images of the apical, middle, and cervical thirds were taken before and after the irrigant activation. The Kappa test was used to determine interexaminer agreement. Statistical analysis was performed using Kruskal-Wallis, Mann-Whitney, and Wilcoxon tests (p < 0.05). PUI and EC improved the removal of remnant filling material in all root canal thirds (p < 0.05). PUI and EC presented similar performance in the final step of retreatment (p > 0.05). No significant difference was observed in the removal of filling material in the apical, middle, and cervical thirds in both groups (p > 0.05). EasyClean in continuous rotary motion is useful in retreatment and was shown to be as effective as ultrasonic activation in the removal of remnant filling material.

Sealing ability of MTA and radiopaque Portland cement with or without calcium chloride for root-end filling.

The aim of this study was to evaluate the influence of calcium chloride (CaCl(2)) on the sealing ability of three Mineral Trioxide Aggregate (MTA) cements, ProRoot MTA, MTA-Angelus, and radiopaque White Portland cement (WPC), for retrograde root filling. Seventy roots of extracted single-rooted teeth were instrumented and obturated. After sectioning the samples at 2 mm from the apex, they received one layer of Araldite and two coats of nail enamel, except for the apical dentinal surface submitted to apicectomy. Standardized retrograde cavities were prepared, filled with one of the materials, and immersed in 0.2% Rhodamine B solution for 72 hours. Dye leakage was analyzed on a light microscope with ocular micrometer. Kruskal-Wallis and Miller tests were used to compare groups arranged in increasing order of leakage, according to mean rank of scores: WPC+CaCl(2), MTA-Angelus+CaCl(2), ProRoot MTA+CaCl(2), MTA-Angelus, ProRoot MTA, and WPC. CaCl(2) improved the sealing ability of all three MTA cements.

Efficacy of CM-Wire, M-Wire, and Nickel-Titanium Instruments for Removing Filling Material from Curved Root Canals: A Micro-Computed Tomography Study.

INTRODUCTION: The aim of this ex vivo study was to evaluate the removal of filling material after using CM-wire, M-wire, and nickel-titanium instruments in both reciprocating and rotary motions in curved canals. METHODS: Thirty maxillary lateral incisors were divided into 9 groups according to retreatment procedures: Reciproc R25 followed by Mtwo 40/.04 and ProDesign Logic 50/.01 files; ProDesign R 25/.06 followed by ProDesign Logic 40/.05 and ProDesign Logic 50/.01 files; and Gates-Glidden drills, Hedström files, and K-files up to apical size 30 followed by K-file 40 and K-file 50 up to the working length. Micro-computed tomography scans were performed before and after each reinstrumentation procedure to evaluate root canal filling removal. Statistical analysis was performed with Kruskal-Wallis, Friedman, and Wilcoxon tests (P < .05). RESULTS: No significant differences in filling material removal were found in the 3 groups of teeth. The use of Mtwo and ProDesign Logic 40/.05 rotary files did not enhance filling material removal after the use of reciprocating files. The use of ProDesign Logic 50/.01 files significantly reduced the amount of filling material at the apical levels compared with the use of reciprocating files. CONCLUSIONS: Association of reciprocating and rotary files was capable of removing a large amount of filling material in the retreatment of curved canals, irrespective of the type of alloy of the instruments. The use of a ProDesign Logic 50/.01 file for apical preparation significantly reduced the amount of remnant material in the apical portion when compared with reciprocating instruments.

Prevalence and morphometric analysis of three-rooted mandibular first molars in a Brazilian subpopulation.

OBJECTIVES:: To determine the prevalence of three-rooted mandibular molars in a Brazilian population using cone beam computed tomography (CBCT) and to analyze the anatomy of mandibular first molars with three roots through micro-CT. MATERIAL AND METHODS:: CBCT images of 116 patients were reviewed to determine the prevalence of three-rooted first mandibular molars in a Brazilian subpopulation. Furthermore, with the use of micro-CT, 55 extracted three-rooted mandibular first molars were scanned and reconstructed to assess root length, distance between canal orifices, apical diameter, Vertucci's classification, presence of apical delta, number of foramina and furcations, lateral and accessory canals. The distance between the orifice on the pulp chamber floor and the beginning of the curvature and the angle of canal curvature were analyzed in the distolingual root. Data were compared using the Kruskal-Wallis test (α=0.05). RESULTS:: The prevalence of three-rooted mandibular first molars was of 2.58%. Mesial roots showed complex distribution of the root canal system in comparison to the distal roots. The median of major diameters of mesiobuccal, mesiolingual and single mesial canals were: 0.34, 0.41 and 0.60 mm, respectively. The higher values of major diameters were found in the distobuccal canals (0.56 mm) and the lower diameters in the distolingual canals (0.29 mm). The lowest orifice distance was found between the mesial canals (MB-ML) and the highest distance between the distal root canals (DB-DL). Almost all distal roots had one root canal and one apical foramen with few accessory canals. CONCLUSIONS:: Distolingual root generally has short length, severe curvature and a single root canal with low apical diameter.

Sealing ability of MTA, CPM, and MBPc as root-end filling materials: a bacterial leakage study.

Objectives To evaluate the sealing ability of three root-end filling materials (white MTA, CPM, and MBPc) using an Enterococcus faecalis leakage model. Material and Methods Seventy single-root extracted human teeth were instrumented and root-ends were resected to prepare 3 mm depth cavities. Root-end preparations were filled with white MTA, CPM, and MBPc cements. Enterococcus faecalis was coronally introduced and the apical portion was immersed in BHI culture medium with phenol red indicator. The bacterial leakage was monitored every 24 h for 4 weeks. The statistical analysis was performed using the Wilcoxon-Gehan test (p<0.05). Results All cements showed bacterial leakage after 24 hours, except for the negative control group. The MBPc showed significantly less bacterial leakage compared with the MTA group (p<0.05). No significant differences were found between the CPM and the other groups. Conclusions The epoxy resin-based cement MBPc had lower bacterial leakage compared with the calcium silicate-based cements MTA and CPM.

Finishing of glass ionomer retroseals.

Various techniques have been suggested for the finishing of retrosealing materials. By measuring marginal leakage and by the use of scanning electronic microscopy, this study evaluated the use of bone files, burnishers, and Shofu points for the finishing of 60 retrosealings done with glass ionomer. With each of the three finishing agents, one group of teeth was polished with movements directed from the palatal toward the buccal, and in another group it was done from the center of the retroseal toward the margins of the cavity. ANOVA showed significant differences among the groups according to the way the finishing was done. The use of instruments from the center to the margins of the cavity rather than in just one direction always caused less leakage. The Shofu stone and the burnisher caused equivalent leakage which was significantly less than that caused by bone files. Scanning electron microscopic photos show that the use of the instrument in only one direction caused the material to shift away from the palatal side. This shift always led to greater leakage of dyes on that side.

Micro-computed tomography analysis of the root canal anatomy and prevalence of oval canals in mandibular incisors.

INTRODUCTION: This study aimed to describe the anatomy of the mandibular incisors by using micro-computed tomography. METHODS: Mandibular incisors (n = 340) were scanned at 19-µm voxel size resolution, and the numbers of canals were classified according to Vertucci classification, as well as the major and minor diameters of the root and root canals, presence of oval canals, and three-dimensional analysis of the apical third were also measured. Data were presented in terms of median and range for each anatomic classification. RESULTS: Overall, the specimens had 1 root canal (N = 257). The second most prevalent anatomy was Vertucci type III (N = 56). These anatomies represent 92% of the sample. The medians of the major diameter at the 1-, 2-, and 3-mm level of the most prevalent anatomies were 0.36, 0.39, and 0.47 mm for type I and 0.41, 0.51, and 0.66 mm for type III, respectively. The apical volume appears to be constant among these anatomies (0.63 and 0.59 mm(3)). Oval canals were found at the 1-mm apical level, with a prevalence of 16.7% for Vertucci type I and 37.5% for Vertucci type III. The presence of oval canals increased at the 3-mm apical level to 32.4% and 76.2% for Vertucci type I and III classifications, respectively. CONCLUSIONS: Type I and III configurations represent 92% of the mandibular incisors studied. Within these anatomic configurations, oval-shaped canals in the apical third were not uncommon and more prevalent in the type III anatomy.

Biocompatibility and setting time of CPM-MTA and white Portland cement clinker with or without calcium sulfate.

OBJECTIVE: To evaluate the biocompatibility and the setting time of Portland cement clinker with or without 2% or 5% calcium sulfate and MTA-CPM. MATERIAL AND METHODS: Twenty-four mice (Rattus norvegicus) received subcutaneously polyethylene tubes filled with Portland cement clinker with or without 2% or 5% calcium sulfate and MTA. After 15, 30 and 60 days of implantation, the animals were killed and specimens were prepared for microscopic analysis. For evaluation of the setting time, each material was analyzed using Gilmore needles weighing 113.5 g and 456.5 g, according to the ASTM specification Number C266-08 guideline. Data were analyzed by ANOVA and Tukey's test for setting time and Kruskal-Wallis and Dunn test for biocompatibility at 5% significance level. RESULTS: Histologic observation showed no statistically significant difference of biocompatibility (p>0.05) among the materials in the subcutaneous tissues. For the setting time, clinker without calcium sulfate showed the shortest initial and final setting times (6.18 s/21.48 s), followed by clinker with 2% calcium sulfate (9.22 s/25.33 s), clinker with 5% calcium sulfate (10.06 s/42.46 s) and MTA (15.01 s/42.46 s). CONCLUSIONS: All the tested materials showed biocompatibility and the calcium sulfate absence shortened the initial and final setting times of the white Portland cement clinker.

Antimicrobial activity of triantibiotic paste, 2% chlorhexidine gel, and calcium hydroxide on an intraoral-infected dentin biofilm model.

INTRODUCTION: The purpose of this study was to evaluate the antimicrobial activity of calcium hydroxide, 2% chlorhexidine gel, and triantibiotic paste (ie, metronidazole, minocycline, and ciprofloxacin) by using an intraorally infected dentin biofilm model. METHODS: Forty bovine dentin specimens were infected intraorally using a removable orthodontic device in order to induce the biofilm colonization of the dentin. Then, the samples were treated with the medications for 7 days. Saline solution was used as the control. Two evaluations were performed: immediately after the elimination of the medication and after incubation in brain-heart infusion medium for 24 hours. The Live/Dead technique (Invitrogen, Eugene, OR) and a confocal microscope were used to obtain the percentage of live cells. Nonparametric statistical tests were performed to show differences in the percentage of live cells among the groups (P < .05). RESULTS: Calcium hydroxide and 2% chlorhexidine gel did not show statistical differences in the immediate evaluation. However, after application of the brain-heart infusion medium for 24 hours, 2% gel chlorhexidine showed a statistically lesser percentage of live cells in comparison with calcium hydroxide. The triantibiotic paste significantly showed a lower percentage of live cells in comparison with the 2% chlorhexidine gel and calcium hydroxide groups in the immediate and secondary (after 24 hours) evaluations. CONCLUSIONS: The triantibiotic paste was most effective at killing the bacteria in the biofilms on the intraorally infected dentin model in comparison with 2% chlorhexidine gel and calcium hydroxide.

Antimicrobial activity of calcium hydroxide and chlorhexidine on intratubular Candida albicans.

This study investigated the efficacy of calcium hydroxide and chlorhexidine gel for the elimination of intratubular Candida albicans (C. albicans). Human single-rooted teeth contaminated with C. albicans were treated with calcium hydroxide, 2% chlorhexidine gel, calcium hydroxide plus 2% chlorhexidine gel, or saline (0.9% sodium chloride) as a positive control. The samples obtained at depths of 0-100 and 100-200 µm from the root canal system were analyzed for C. albicans load by counting the number of colony forming units and for the percentage of viable C. albicans using fluorescence microscopy. First, the antimicrobial activity of calcium hydroxide and the 2% chlorhexidine gel was evaluated by counting the number of colony forming units. After 14 days of intracanal medication, there was a significant decrease in the number of C. albicans colony forming units at a depth of 0-100 µm with chlorhexidine treatment either with or without calcium hydroxide compared with the calcium hydroxide only treatment. However, there were no differences in the number of colony forming units at the 100-200 µm depth for any of the medications investigated. C. albicans viability was also evaluated by vital staining techniques and fluorescence microscopy analysis. Antifungal activity against C. albicans significantly increased at both depths in the chlorhexidine groups with and without calcium hydroxide compared with the groups treated with calcium hydroxide only. Treatments with only chlorhexidine or chlorhexidine in combination with calcium hydroxide were effective for elimination of C. albicans.

Influence of root canal sealer on the radiographic appearance of filling voids in maxillary single-rooted teeth.

OBJECTIVE: This study compared the influence of three epoxy resin-based sealers with distinct radiopacities on the observers' ability to detect root canal filling voids during radiographic analysis. MATERIAL AND METHODS: The root canals of 48 extracted maxillary canines were prepared and divided into three groups. Each group was laterally condensed with one sealer (AH Plus®, Acroseal® or a non-radiopaque sealer), and a longitudinal void was simulated in half of the specimens from each group (n=8). Buccolingual radiographs were obtained and randomly interpreted for voids by a radiologist and an endodontist in a blinded fashion. Teeth were cut and inspected under a microscope to confirm the position of void. Differences in sensitivity and specificity between groups and examiners were compared using the Fisher's Exact and McNemar tests, respectively (α=0.05). RESULTS: Significantly lower sensitivity levels (p<0.05) were observed in the coronal portion of fillings performed with both radiopaque sealers. Specificity values for Acroseal® were significantly higher (p<0.05) in the coronal and apical portions of fillings. CONCLUSIONS: The type of root canal sealer can affect the observers' ability to detect root canal filling voids during radiographic analysis of upper single-rooted teeth.