Brain oxidative stress mediates anxiety-like behavior induced by indomethacin in zebrafish: protective effect of alpha-tocopherol.

RATIONALE: Indomethacin (INDO) is a widely utilized non-steroidal anti-inflammatory drug (NSAID) with recognized effect on the central nervous system. Although previous reports demonstrate that prolonged treatment with indomethacin can lead to behavioral alterations such as anxiety disorder, the biochemical effect exerted by this drug on the brain are not fully understood. OBJECTIVES: The aim of present study was to evaluate if anxiety-like behavior elicited by indomethacin is mediated by brains oxidative stress as well as if alpha-tocopherol, a potent antioxidant, is able to prevent the behavioral and biochemical alterations induced by indomethacin treatment. METHODS: Zebrafish were utilized as experimental model and subdivided into control, INDO 1 mg/Kg, INDO 2 mg/Kg, INDO 3 g/Kg, α-TP 2 mg/Kg, α-TP 2 mg/Kg + INDO 1 mg/Kg and α-TP + INDO 2 mg/Kg groups. Vertical distributions elicited by novelty and brain oxidative stress were utilized to determinate behavioral and biochemical alterations elicited by indomethacin treatment, respectively. RESULTS: Our results showed that treatment with indomethacin 3 mg/kg induces animal death. No changes in animal survival were observed in animals treated with lower doses of indomethacin. Indomethacin induced significant anxiogenic-like behavior as well as intense oxidative stress in zebrafish brain. Treatment with alpha-tocopherol was able to prevent anxiety-like behavior and brain oxidative stress induced by indomethacin. CONCLUSIONS: Data presented in current study demonstrated for the first time that indomethacin induces anxiety-like behavior mediated by brain oxidative stress in zebrafish as well as that pre-treatment with alpha-tocopherol is able to prevent these collateral effects.

The impact of long-term storage conditions on the development of experimental dentinal microcracks.

This study evaluated the impact of long-term storage conditions (medium and time) on the development of experimental dentinal microcracks through micro-computed tomography. Sixty freshly extracted premolars were stored in formalin, water, or dry conditions (n = 20) and scanned after 72 h, 30 days, 6 months and 3 years of extraction. The effect of the storage medium and time on the occurrence of dentinal defects was statistically evaluated. A total of 211 000 images were screened revealing the existence of 11 519 slices with dentinal defects. Dry conditions significantly contributed to the development of new defects in all time points. During the 3-year follow-up period, no new defects were detected in the teeth that were stored in water and, in a single tooth, in the formalin group, after 6 months of storage.

Success rate of permanent teeth pulpotomy using bioactive materials: A systematic review and meta-analysis of randomized clinical trials.

BACKGROUND: The results of vital pulp treatments in permanent teeth have been encouraging. Currently, pulpotomy treatment for permanent teeth primarily utilizes mineral trioxide aggregate (MTA) as the dressing material, followed by calcium hydroxide. While other calcium-silicate-based cements have been suggested for pulpotomy, there is a limited number of studies evaluating their long-term effectiveness. OBJECTIVES: The objective of this systematic review and meta-analysis was to evaluate the success rate of pulpotomies performed on permanent teeth, comparing the use of ProRoot MTA with that of calcium hydroxide and other bioceramic materials. METHODS: A comprehensive search was conducted in several electronic databases, including PubMed, Cochrane Library, Scopus, Web of Science, Embase and Science Direct until December 2022. The search was guided by PICOS criteria, including only randomized clinical trials (RCTs) that evaluated the success rate of pulpotomy treatments in permanent teeth using ProRoot MTA in comparison to calcium hydroxide and other bioceramic materials. The quality of the included studies was assessed using the RoB-2 tool to evaluate the risk of bias, and relevant data were extracted and analysed in RevMan software 5.3 using fixed-effect models. The GRADE tool was used to determine the overall quality of evidence. RESULTS: The initial search retrieved 1072 studies and, after eliminating duplicates, 677 studies were screened and 28 studies were considered for eligibility. In the final selection process, 16 studies were included in the systematic review, with 10 being determined as having a high risk of bias. Pulpotomy showed an overall mean success rate of 92% after 1 year. The meta-analysis indicated a significantly higher success rate for pulpotomies utilizing MTA in comparison with calcium hydroxide, while no significant difference was seen between MTA and calcium-enriched mixture (CEM) or Biodentine. The GRADE assessment revealed an overall low level of evidence for the included studies. DISCUSSION: Most randomized controlled trials exhibited a significant absence of control over confounding factors. CONCLUSIONS: This systematic review and meta-analysis demonstrate that pulpotomy is a highly effective treatment for managing permanent teeth. The results indicate that the success rate of pulpotomy using ProRoot MTA is significantly higher than when using calcium hydroxide. However, the certainty of evidence supporting these findings is low, and there is a need for well-designed RCTs to assess the long-term outcomes of pulpotomy using newer bioceramic materials. REGISTRATION: This systematic review was registered in the PROSPERO database (registration number CRD42023393970).

Volumetric changes in root canal sealers in ex vivo and a novel animal model approach.

AIM: This study aimed to evaluate the volumetric change of root canal sealers through micro-computed tomographic analysis using a novel in vivo model and to compare the results with those obtained using an ex vivo test. METHODOLOGY: Eighteen single-rooted teeth were cut to 5 mm length from the root apex. The root canals were uniformly enlarged and filled with EndoSequence BC Sealer or AH Plus Jet root canal sealers. Samples were stored at 37°C and 95% relative humidity for 24 h and then scanned with a micro-CT device. Twelve samples (n = 6 for each sealer) were implanted in the subcutaneous tissue of Wistar rats, while six samples (n = 3 for each sealer) were immersed in 20 mL of phosphate-buffered saline (PBS) at 37°C at neutral pH. After 7 and 30 days, teeth were removed from subcutaneous tissue or PBS and rescanned. Statistical analysis of volume changes was performed using Shapiro-Wilk's test and independent t-test (p < .05). RESULTS: AH Plus Jet had smaller volume changes (-2.2 to +0.77%) than EndoSequence BC Sealer (-2.0 to +4.0%) (p < .05), in the two tested models. The volume of the root canal sealers decreased over time (p < .05), in vivo. AH Plus Jet results varied between the in vivo and ex vivo results (p < .05), while EndoSequence BC Sealer presented similar volume losses for both experimental models (p > .05). CONCLUSION: EndoSequence BC Sealer lost more volume than AH Plus Jet. The experimental conditions influenced the volumetric change of AH Plus Jet but not the EndoSequence BC Sealer. The ex vivo model should be further explored as a methodological alternative to assess the volumetric changes of root canal sealers without causing harm to animals.

Influence of bioceramic intracanal medication on the bond strength of bioceramic root canal sealer.

To investigate the influence of the remaining volume of a new intracanal medication based on bioceramic compounds on the bond strength (BS) and formation of an adhesive interface between calcium silicate-based and epoxy resin-based root canal sealers. For this purpose, the specimens were distributed according to the intracanal medication (n = 26): Bio-C Temp (BCT) and Ultracal XS (UXS). The roots were scanned in microCT, and after 7 days, the medication was removed. Then a new scan was performed to evaluate the volume of medication remaining. Subsequently, 40 specimens were redistributed into 2 subgroups (n = 10) and filled according to the sealer used: AH Plus (AHP) and Bio-C Sealer (BCS), to assess the bond strength by using the push-out test, and the adhesive interface by confocal laser fluorescence microscopy (CLSM) and scanning electron microscopy (SEM). The t test showed a smaller remainder of BCT (1.77 ± 0.86) compared with UXS (10.47 ± 5.78), irrespective of the root third evaluated. The BS showed that teeth with BCT + BCS had higher bond strength values (3.70 ± 1.22) when compared to the other groups: BCT + AHP (2.15 ± 1.07), UXS + BCS (3.18 ± 1.09) and UXS + AHP (2.11 ± 1.02) (p<0.001). The cervical third had higher BS when compared with the middle and apical thirds (p < 0.001), and higher number of adhesive failures. The adhesive interface in SEM and CLSM images showed better adaptation for the association between BCT + BCS. Intracanal medication and silicate-based endodontic sealer appeared to interact chemically by forming a biomineralizing layer, allowing for an increase in the bond strength and forming an adhesive interface between the materials, with no or less gap formation.

Clinical Accuracy and Precision of 3 Multifrequency Electronic Apex Locators Assessed through Micro-Computed Tomographic Imaging.

INTRODUCTION: This study aimed to compare the in vivo accuracy and precision of 3 electronic apex locators (EALs) in determining the position of the major foramen using micro-computed tomographic (micro-CT) technology. METHODS: After access preparation of 23 necrotic or vital teeth from 5 patients, canals were negotiated, and hand files were used to determine the position of the foramen with 3 EALs: Propex Pixi (Dentsply Maillefer, Ballaigues, Switzerland), Woodpex III (Woodpecker Medical Instrument Co, Guilin, China), and Root ZX II (J Morita, Tokyo, Japan). After fixing the silicon stop to the file, teeth were extracted and scanned in a micro-CT device with and without the instrument inserted into the canal. Data sets were coregistered, and the accuracy and precision of the EALs were determined at a tolerance level of ±0.5 mm by measuring the distance from the tip of the instruments to a tangential line crossing the margins of the foramen. Statistical comparisons were performed using Friedman with post hoc related samples sign and Spearman tests (α = 5%). RESULTS: A significant difference was detected comparing the accuracy of Root ZX II (100%), Woodpex III (86.96%), and Propex Pixi (52.17%) (P < .05). There was a lack of significance in the relationship between the pulp status and the accuracy of the tested EALs (P > .05). Propex Pixi was significantly less precise than Root ZX II (P < .05), whereas no difference was found between Woodpex III and Root ZX II or Propex Pixi (P > .05). CONCLUSIONS: EALs presented similar precision, but Woodpex III and Root ZX II showed better accuracy to determine the position of the apical major foramen than Propex Pixi.

Taper 0.06 Versus Taper 0.04: The Impact on the Danger Zone.

INTRODUCTION: This study aimed to evaluate the effects of root canal enlargement on the danger zone (DZ) of mandibular molars. METHODS: Thirty mesial roots of mandibular first molars were scanned in micro-computed tomography (S1). Canals were sequentially enlarged with rotary instruments up to sizes 30/0.04 (S2) and 30/0.06 (S3). The dentin thickness was measured at 0.1-mm intervals after each preparation step (n = 2964 slices). Root level and position of the DZ were also recorded. Data were compared using analysis of variance with Bonferroni pairwise comparison, Cochran's Q method, and Pearson's test (α = 5%). RESULTS: Comparing the specimens before (S1) and after (S2 and S3) preparations showed a significant reduction in the thickness of the DZ (P < .05), as well as between S2 and S3 steps (P < .05). At S1, the DZ was mostly located in the middle third of the root, but after preparation, it shifted toward the coronal direction (P < .05). Both S2 (P = .004, r = 0.508) and S3 (P = .004, r = 0.506) preparation steps showed a positive correlation between canal length and the root level of the DZ. At S1, the DZ was positioned toward the distal and mesial in 73.4% (n = 22) and 26.6% (n = 8) of the specimens, respectively. After S3, the number of specimens with DZ positioned toward the mesial aspect of the root significantly reduced to 3.3% (n = 1), whereas none of the specimens with DZ positioned toward the distal changed its position after root canal enlargements (P > .05). CONCLUSION: Overall, the enlargement of mesial canals of mandibular first molars with final instruments sizes 30/0.04 and 30/0.06 affected the thickness, root level, and position of the DZ.

Regenerative Endodontics by Cell Homing: A Review of Recent Clinical trials.

INTRODUCTION: The conventional treatment for irreversibly inflamed or necrotic teeth is root canal treatment or apexification. Regenerative endodontics aims to regenerate the damaged "pulp-like" tissue, which can preserve the teeth' vitality and sensitivity while avoiding necrosis. The main clinical benefit is root maturation. The "pulp-like" tissue does not refer to regenerated pulp tissue with an odontoblastic layer or the formation of pulp-dentin complexes. The cell homing technique is built on endogenous stem cells and their capacity to regenerate tissue. Cell homing refers to endogenous cells' migration or infiltration into the cite when stimulated by physiochemical or biological stimuli or by passive flow with a blood clot from the apical tissue. Its Regenerative Endodontic Procedures success criteria are defined by the American Association of Endodontists. The purpose of this article is to provide an overview of vital pulp tissue and various strategies to promote regeneration of damaged pulp tissue. The cell homing technique will be reviewed through clinical trials. METHODS: We performed a comprehensive literature review on a total of nine clinical trials of regenerative endodontics using the cell-homing technique based on three databases and duplicate manuscripts were removed. RESULTS: Regenerative endodontics using the cell-homing technique shows promising results that can be translated into clinical practice. However, a favorable result was observed in immature teeth, and the results are contradictory in mature teeth. CONCLUSION: Regeneration therapy is an attractive new alternative to conventional endodontic treatments. Preservation of vitality and continuation of root development in damaged teeth would be a clear advantage.

Influence of bioceramic intracanal medication on the bond strength of bioceramic root canal sealer

Abstract To investigate the influence of the remaining volume of a new intracanal medication based on bioceramic compounds on the bond strength (BS) and formation of an adhesive interface between calcium silicate-based and epoxy resin-based root canal sealers. For this purpose, the specimens were distributed according to the intracanal medication (n = 26): Bio-C Temp (BCT) and Ultracal XS (UXS). The roots were scanned in microCT, and after 7 days, the medication was removed. Then a new scan was performed to evaluate the volume of medication remaining. Subsequently, 40 specimens were redistributed into 2 subgroups (n = 10) and filled according to the sealer used: AH Plus (AHP) and Bio-C Sealer (BCS), to assess the bond strength by using the push-out test, and the adhesive interface by confocal laser fluorescence microscopy (CLSM) and scanning electron microscopy (SEM). The t test showed a smaller remainder of BCT (1.77 ± 0.86) compared with UXS (10.47 ± 5.78), irrespective of the root third evaluated. The BS showed that teeth with BCT + BCS had higher bond strength values (3.70 ± 1.22) when compared to the other groups: BCT + AHP (2.15 ± 1.07), UXS + BCS (3.18 ± 1.09) and UXS + AHP (2.11 ± 1.02) (p<0.001). The cervical third had higher BS when compared with the middle and apical thirds (p < 0.001), and higher number of adhesive failures. The adhesive interface in SEM and CLSM images showed better adaptation for the association between BCT + BCS. Intracanal medication and silicate-based endodontic sealer appeared to interact chemically by forming a biomineralizing layer, allowing for an increase in the bond strength and forming an adhesive interface between the materials, with no or less gap formation.

Mechanical performance of original; yellowish and blueish ProFile instruments: isolating heat-treatment as a variable.

The present study aimed to perform two different heat-treatments in an austenitic NiTi ProFile instrument and to compare the mechanical performance of original and heat-treated instruments. Heat treatment of ProFile (tip size 25 and 0.06 taper) instruments were carried out in a furnace in argon atmosphere using a heating rate of 10° C/min. After reaching the programmed temperatures of 450 ºC or 500 ºC the system remained at a constant temperature for 10 minutes; followed by cooling in water at room temperature. Afterwards; the three groups (n=30 per group) of instruments were compared regarding their cyclic fatigue (n=10 per group); bending (n=10 per group); and buckling resistance (n=10 per group). After cyclic fatigue tests; a scanning electron microscope was used to analyze the fracture surfaces and observe the fracture mode. Statistical analysis was performed using One-way ANOVA and Student-Newman-Keuls test; with an alpha type error set at 0.05. Yellowish and blueish coloration was observed in the ProFile instruments after 450 ºC or 500 ºC heat treatments; respectively. Conventional ProFile instruments showed the lower cyclic fatigue; and the higher bending and buckling resistance (P<0.05). In contrast; yellowish ProFile instruments (heat treated at 500° C) showed the higher cyclic fatigue; and the lower bending and buckling resistance (P>0.05). It can be concluded that the different heat treatments performed on ProFile instruments increased its cyclic fatigue resistance and improved the flexibility and buckling resistance.

O presente estudo teve como objetivo realizar dois tratamentos térmicos diferentes em instrumentos de NiTi austenítico ProFile e comparar o desempenho mecânico de instrumentos originais e tratados termicamente. O tratamento térmico dos instrumentos ProFile (tamanho de ponta 25 e conicidade 0.06) foi realizado em um forno em atmosfera de argônio usando uma taxa de aquecimento de 10°C/min. Após atingir as temperaturas programadas de 450 ºC ou 500 ºC o sistema permaneceu em temperatura constante por 10 minutos; seguido de resfriamento em água à temperatura ambiente. Em seguida; os três grupos de instrumentos (n=30 por grupo) foram comparados quanto à fadiga cíclica (n=10 por grupo); flexão (n=10 por grupo) e resistência à flambagem (n=10 por grupo). Após testes de fadiga cíclica; um microscópio eletrônico de varredura foi utilizado para analisar as superfícies de fratura e observar o modo de fratura. A análise estatística foi realizada por meio dos testes de One-way ANOVA e teste de Student-Newman-Keuls; com erro de tipo alfa fixado em 0.05. Coloração amarelada e azulada foi observada nos instrumentos ProFile após tratamentos térmicos a 450 ºC ou 500 ºC; respectivamente. Os instrumentos ProFile originais apresentaram menor fadiga cíclica e maior resistência à flexão e flambagem (P<0.05). Em contraste; os instrumentos ProFile azulados (tratados termicamente a 500°C) apresentaram maior fadiga cíclica e menor resistência à flexão e flambagem (P>0.05). Pode-se concluir que os diferentes tratamentos térmicos realizados nos instrumentos ProFile aumentaram sua resistência à fadiga cíclica e melhoraram a flexibilidade e resistência à flambagem.

Mechanical performance of original; yellowish and blueish ProFile instruments: isolating heat-treatment as a variable

Resumo O presente estudo teve como objetivo realizar dois tratamentos térmicos diferentes em instrumentos de NiTi austenítico ProFile e comparar o desempenho mecânico de instrumentos originais e tratados termicamente. O tratamento térmico dos instrumentos ProFile (tamanho de ponta 25 e conicidade 0.06) foi realizado em um forno em atmosfera de argônio usando uma taxa de aquecimento de 10°C/min. Após atingir as temperaturas programadas de 450 ºC ou 500 ºC o sistema permaneceu em temperatura constante por 10 minutos; seguido de resfriamento em água à temperatura ambiente. Em seguida; os três grupos de instrumentos (n=30 por grupo) foram comparados quanto à fadiga cíclica (n=10 por grupo); flexão (n=10 por grupo) e resistência à flambagem (n=10 por grupo). Após testes de fadiga cíclica; um microscópio eletrônico de varredura foi utilizado para analisar as superfícies de fratura e observar o modo de fratura. A análise estatística foi realizada por meio dos testes de One-way ANOVA e teste de Student-Newman-Keuls; com erro de tipo alfa fixado em 0.05. Coloração amarelada e azulada foi observada nos instrumentos ProFile após tratamentos térmicos a 450 ºC ou 500 ºC; respectivamente. Os instrumentos ProFile originais apresentaram menor fadiga cíclica e maior resistência à flexão e flambagem (P<0.05). Em contraste; os instrumentos ProFile azulados (tratados termicamente a 500°C) apresentaram maior fadiga cíclica e menor resistência à flexão e flambagem (P>0.05). Pode-se concluir que os diferentes tratamentos térmicos realizados nos instrumentos ProFile aumentaram sua resistência à fadiga cíclica e melhoraram a flexibilidade e resistência à flambagem.

Abstract The present study aimed to perform two different heat-treatments in an austenitic NiTi ProFile instrument and to compare the mechanical performance of original and heat-treated instruments. Heat treatment of ProFile (tip size 25 and 0.06 taper) instruments were carried out in a furnace in argon atmosphere using a heating rate of 10° C/min. After reaching the programmed temperatures of 450 ºC or 500 ºC the system remained at a constant temperature for 10 minutes; followed by cooling in water at room temperature. Afterwards; the three groups (n=30 per group) of instruments were compared regarding their cyclic fatigue (n=10 per group); bending (n=10 per group); and buckling resistance (n=10 per group). After cyclic fatigue tests; a scanning electron microscope was used to analyze the fracture surfaces and observe the fracture mode. Statistical analysis was performed using One-way ANOVA and Student-Newman-Keuls test; with an alpha type error set at 0.05. Yellowish and blueish coloration was observed in the ProFile instruments after 450 ºC or 500 ºC heat treatments; respectively. Conventional ProFile instruments showed the lower cyclic fatigue; and the higher bending and buckling resistance (P<0.05). In contrast; yellowish ProFile instruments (heat treated at 500° C) showed the higher cyclic fatigue; and the lower bending and buckling resistance (P>0.05). It can be concluded that the different heat treatments performed on ProFile instruments increased its cyclic fatigue resistance and improved the flexibility and buckling resistance.

Micro-CT Study of the In Vivo Accuracy of a Wireless Electronic Apex Locator.

INTRODUCTION: This study aimed to compare the in vivo accuracy of Wirele-X and RootZX II electronic apex locators (EALs) in determining the position of the major foramen using micro-computed tomography (micro-CT) as the analytical tool. METHODS: Eleven vital teeth planned for extraction from 5 patients were used. After conventional access cavity preparation, root canals were flared and negotiated up to the apical third with sizes 08 and 10 K-files followed by irrigation with 2.5% NaOCl. K-type files were used to determine the working length of the selected canals using Root ZX II and Wirele-X apex locators until their numeric displays read "0.0." After fixing the silicon stop to the file, teeth were extracted and imaged in a micro-CT device using a double-scan protocol. Image stacks, with and without the file in the root canal, were then co-registered and the measurement error calculated as the absolute difference between the tip of the file and the major foramen. Positive and negative values were recorded when the file tip was detected beyond or short of the major foramen, respectively. Accuracy was determined on stable measurements within ± 0.5 mm when the file tip did not extend beyond the major foramen. The χ2 test was applied to compare the ability of the EALs to detect the position of the major foramen, and the t test for dependent variables was used to verify differences in the 2 measurements obtained in each tooth. Significance level was set at 5%. RESULTS: Within a tolerance level of ± 0.5 mm, no significant differences were observed between the tested EALs regarding the absolute distance values (P = .82) or in their ability to detect the position of the major foramen (χ2 = 0.2588; P = .6109). The accuracy of the Root ZX II and the Wirele-X apex locators within ± 0.5 mm were 81.8% and 90.9%, respectively. CONCLUSIONS: Root ZX II and Wirele-X performed similarly regarding the in vivo detection of the major foramen. Using strict criteria, the accuracy of the Root ZX II and the Wirele-X apex locators were 81.8% and 90.9%, respectively.

Micro-CT assessment of gap-containing areas along the gutta-percha-sealer interface in oval-shaped canals.

AIM: To assess the interfacial adaptation (gap-containing areas) of two root canal sealers (EndoSequence BC Sealer and AH Plus) to a conventional gutta-percha (GP) cone (GP-sealer interface) in oval-shaped canals filled using the single cone technique. METHODOLOGY: Sixteen teeth with oval-shaped canals were selected, scanned in a micro-computed tomographic device, and pair-matched according to the volume, aspect ratio and configuration of root canals. Root canals were then sequentially prepared with WaveOne Gold Primary and Large instruments, followed by filling with WaveOne Large GP points associated with either the premixed calcium-silicate EndoSequence BC Sealer or the epoxy resin-based AH Plus sealer (n = 8 per group) using the single cone technique. After 7 days stored in phosphate-buffered saline solution at 37°C, the specimens were rescanned and the reconstructed images segmented in order to differentiate the filling materials (sealer and GP cone) to the dentine. A total of 453 cross-sectional slices were assessed and categorized according to the presence or absence of gaps at the GP-sealer interface. Mann-Whitney U-test verified the differences between groups and were considered significant at alpha = 5%. RESULTS: Gaps were non-homogenously distributed in two-dimensional axial cross-section images and none of the specimens showed completely gap-free areas along the entire GP-sealer interface. Root canals filled with EndoSequence BC Sealer and AH Plus displayed 171 (37.75%) and 136 (30.02%) slices with gaps in the GP-sealer interface and these frequencies were statistically significant (p = .000). CONCLUSIONS: Although none of the specimens had a gap-free area along the entire GP-sealer interface, oval canals filled with AH Plus showed less gaps than the ones filled with EndoSequence BC Sealer.

A critical analysis of research methods and experimental models to study root canal fillings.

Canal filling materials and techniques have been one of the most studied topics in Endodontics. A simple search using the mesh term "root canal filling" in PubMed revealed more than 11 000 articles, an impressive number that is much higher than "root canal disinfection" (5544 articles) or even the popular "root canal preparation" (8527 articles). The overriding importance attributed to root filling procedures is not merely intuitive. It derived from the appealing relevance given by the appearance of the white lines in common radiographs grounded on retrospective clinical data that had identified the quality of a root filling as one of the major causes of treatment failure (lack of healing). Since the publication of the Washington study, impressive efforts have been made for the release of new materials and techniques, as well as, for the development of a plethora of laboratory methods to assess the quality of root filling procedures. This narrative review aims to address and discuss the most relevant laboratory methods to assess the root canal filling. As filling quality improvements have not translated into higher success rates, as reported in longitudinal clinical studies, more than to deliver a simple methodology-based review, this paper aims to present an in-depth critical view on the assessment of laboratory methods used to study the filling materials and techniques. Recent data indicate that the long-term dimensional stability/degradation over time of endodontic sealers plays a central role in the treatment outcome. In this context, laboratory methods should be developed focusing on predicting, at least to some degree, the long-term clinical behaviour of root canal fillings, rather than simply ranking different materials or techniques.

Present status and future directions - Minimal endodontic access cavities.

In the last decades, the move of medicine towards minimally invasive treatments is notorious and scientifically grounded. As dentistry naturally follows in its footsteps, minimal access preparation have also becume a trend topic in the endodontic field. This procedure aims to maximize preservation of dentine tissue backed up by the idea that this is an effective way to reduce the incidence of post-treatment tooth fracture. However, with the assessment of the body of evidence on this topic, it is possible to observe some key points (a) the demand for nomenclature standardization, (b) the requirement of specific tools such as ultra-flexible instruments, visual magnification, superior illumination, and three-dimensional imaging technology, (c) minimally invasive treatment does not seem to affect orifice location and mechanical preparation when using adequate armamentarium, but it (d) may impair adequate canal cleaning, disinfection and filling procedures, and also (e) it displays contradictory results regarding the ability to increase the tooth strengthen compared to the traditional access cavity. In spite of that, it is undeniable that methodological flaws of some benchtop studies using extracted teeth may be responsible for the conflicting data, thus triggering the need for more sophisticated devices/facilities and specifically designed research in an attempt to make clear the role of the access size/design on long-term teeth survival. Moreover, it is inevitable that a clinical approach such as minimal endodontic access cavities that demands complex tools and skilled and experienced operators bring to the fore doubts on its educational impact mainly when confronted with the conflicting scientific output, ultimately provoking a cost-benefit analysis of its implementation as a routine technique. In addition, this review discusses the ongoing scientific and clinical status of minimally invasive access cavities aiming to input an in-depth and unbiased view over the rationale behind them, uncovering not only the related conceptual and scientific flaws but also outlining future directions for research and clinical practices. The conclusions attempt to skip from passionate disputes highlighting the current body of evidence as weak and incomplete to guide decision making, demanding the development of a close-to-in situ laboratory model or a large and well-controlled clinical trial to solve this matter.

Influence of access cavity preparation on the dentine thickness of mesial canals of mandibular molars prepared with reciprocating instruments.

AIM: To evaluate the influence of traditional and conservative access cavity preparations on the remaining dentine thickness in the coronal third of mesial canals of extracted mandibular molars prepared with reciprocating instruments using micro-computed tomography as the analytical tool. METHODOLOGY: Seventy extracted mandibular molars were scanned at a pixel size of 19 µm. From this initial sample, 20 teeth were selected, pair-matched and distributed into two groups (n = 10) according to the access cavity preparation: traditional (TradAC) or conservative (ConsAC). The root canals were sequentially enlarged with Reciproc Blue R25 (size 25, 0.08v taper) and R40 (size 40, 0.06v taper) instruments. A new scan was performed and the postoperative stacks were coregistered with their respective preoperative datasets. A colour-coded cross sections of the roots were created and used to identify and measure the smallest dentine thickness related to both MB and ML canals at 1.0-mm intervals from the furcation level of up to 5 mm in the apical direction, in both mesial and distal aspects of the roots, before and after preparation. The statistical analyses were performed with paired-samples t-test, independent-samples Student T-test and Chi-Square test with a significance level of 5%. RESULTS: At all levels of both groups, dentine thickness before preparation was greater than after preparation (p < .05). No difference in the percentage of dentine reduction was observed between TradAC and ConsAC groups (p > .05), but a significantly greater reduction was observed to the distal aspect of the roots (p < .05). After root canal preparation, dentine thinner than 0.5 mm was observed mostly along the distal aspect of the root (10% to 15%) of the MB and ML canals, with no influence of the access cavity type on its incidence to either mesial (X2  = 1.66; p = .2) or distal (X2  = 0.40; p = .5) directions. In the TradAC group, dentine thickness in most slices was greater than 1.0 mm after preparation (n = 124) whilst, in the ConsAC, it ranged from 0.5 to 1.0 mm (n = 136). CONCLUSION: Traditional or conservative access cavity preparation in extracted mandibular molars did not influence the remaining dentine thickness in the coronal third of mesial canals enlarged with thermomechanically treated nickel-titanium reciprocating instruments.

A critical analysis of research methods and experimental models to study dentinal microcracks.

The purpose of this narrative review was to discuss the scientific milestones that led to the current understanding of the root dentinal microcrack phenomenon based on the interplay between the usage of micro-computed tomography (micro-CT) as an analytical tool alongside a close-to-mouth experimental model. In 2009, reports on the development of dentinal microcracks in extracted teeth after root canal preparation triggered an awareness of the potential for vertical root fractures (VRFs) of endodontically treated teeth could be developed from defects created by the mechanical stress of nickel-titanium preparation systems on dentine. This assumption was taken for granted, even though no cause-effect relationship had been scientifically demonstrated. Since then, several studies using the sectioning method with extracted teeth have been published and the large discrepancy amongst their outcomes soon become evident. Moreover, the high frequency of reported dentinal microcracks largely contrasted with the clinical incidence of VRFs, raising doubts on their methodological reliability. Using micro-CT technology, it was demonstrated by several studies that, in extracted teeth, dentinal defects already existed before the endodontic procedures, indicating that the initial reports framed a non-existing cause-effect relationship between canal preparation and dentinal microcracks. Although these new findings contributed to a better comprehension of this phenomenon, the misconception that microcracks were the starting point for VRFs was only surpassed with a new in situ approach using fresh cadavers. Surprisingly, microcracks were not identified in sound teeth. As a conclusion, dentinal microcracks in extracted teeth can be considered a non-natural occurrence observed only in a laboratory set-up as a consequence of dehydration and storage conditions. Thus, dentinal microcracks shall not be considered as the starting point for VRFs as they do not manifest in non-extracted teeth. Identifying dentinal microcracks as a laboratory phenomenon highlights the impact of recent scientific developments to disclaim the clinical relevance of laboratory-obtained outcomes.

An ultrasonic-based experimental model to evaluate the setting time of endodontic sealers.

AIM: To evaluate an ultrasonic-based experimental model (US), to assess the setting time of AH Plus® , Bio-C Sealer® and MTA Fillapex® , and compare the results obtained with ANSI/ADA specifications (2012). METHODS: To determine the setting time according to the ANSI/ADA specification, moulds measuring 10 mm (diameter) × 2 mm (thickness) were used, and for the US tests a transducer (1 MHz) and an oscilloscope were used. Fourier-transform infrared spectroscopy (FTIR) was used to evaluate the changes on chemical structure at the different setting times found by the US and ANSI/ADA methods. After checking the normal distribution, setting time data were analysed using unilateral analysis of variance with Tukey-HSD post-test to compare the methods. RESULTS: AH Plus® and Bio-C Sealer® had longer mean setting time values for the US method than for the ANSI/ADA (p < .05), whilst for MTA Fillapex® no significant difference was found between the methods (p < .05). FTIR spectra demonstrated that at the setting time determined by the US method, AH Plus® and Bio-C Sealer® were in a more advanced stage of polymerization than at the ANSI/ADA, whilst there was no significant difference for MTA Fillapex® . CONCLUSIONS: The use of US was effective for setting time determination and measured longer values than ANSI/ADA specification for AH Plus® and Bio-C Sealer® , identifying the entire (and not only superficial) setting of the endodontic sealers.

Methodological proposal for evaluation of adhesion of root canal sealers to gutta-percha.

AIM: To compare the bond strength of an epoxy resin-based sealer and two calcium silicate-based sealers (CSS) to gutta-percha discs using a new method. METHODOLOGY: Round discs of gutta-percha (n = 60), measuring 10 mm in diameter and 2 mm in thickness, were placed on a glass plate and a drop of each sealer (AH Plus, EndoSequence BC Sealer and EndoSeal MTA) was placed on their surface. Another identical disc was placed onto the first one and a standardized weight (0.0981 N) applied over them using a specially developed apparatus. Ten samples prepared for each sealer were submitted to a microshear bond strength test accomplished by a specially designed set-up coupled to a universal testing machine. The Kruskal-Wallis test followed by a post hoc procedure was used to compare groups considering the preliminary analysis of the raw data had indicated the nonadherence to a Gaussian distribution (Shapiro-Wilk, p < .05). Alpha error was set at 5%. RESULTS: Overall, no premature failure occurred. All sealers had some degree of adhesiveness to gutta-percha discs but with a significant difference amongst them (Kruskal-Wallis, p = .019). The epoxy resin-based sealer (AH Plus) had significantly higher median shear bond strength values (1.43 MPa; 1.40-1.83) compared to EndoSeal MTA (0.53 MPa; 0.46-0.73) (p = .021) and EndoSequence BC Sealer (0.45 MPa; 0.34-0.46) (p = .023), whilst the lowest median value was observed with EndoSequence BC Sealer (0.45 MPa; 0.34-0.46) (p < .05). CONCLUSIONS: CSS sealers had weaker bonding to gutta-percha when compared to the epoxy resin-based AH Plus sealer. The proposed methodology is an innovative and reproducible method for testing the bond strength of root canal sealers to gutta-percha.

Is canal overinstrumentation able to produce apical root dentinal microcracks in extracted teeth?

AIM: To assess the impact of mechanical overinstrumentation of root canals in extracted teeth on the development of dentinal microcracks in the apical portion of the root by means of micro-computed tomographic (micro-CT) analysis. Root canal preparation short of the canal terminus and at the apical foramen (AF) were used as controls. METHODOLOGY: Twenty 2-rooted maxillary premolars with two canals were selected, scanned in a micro-CT device and the root canals prepared sequentially using Reciproc M-Wire R25 instruments to working lengths: 1 mm short of the AF (AF - 1 mm), at the AF (AF), and overinstrumentation (AF + 1 mm). A micro-CT scan of each specimen was performed after each time point. After reconstruction and co-registration procedures, the images were screened from the cementoenamel junction to the apex (n = 55 352) to identify the presence of dentinal microcracks in the apical third of the root. RESULTS: After the analyses of 55 352 slices, dentinal microcracks were visualized in 12.45% of the images (6892 slices), with 5.73% (3174 slices) in the cervical, 3.57% (1976 slices) in the middle and 3.15% (1740 slices) in the apical third. All dentinal microcracks observed at all time points (AF - 1 mm, AF and AF + 1 mm) were already present in the corresponding images before canal instrumentation. Therefore, no new microcracks were detected, regardless of the working length used for canal instrumentation. CONCLUSION: Reciprocating root canal preparation either short, at or beyond the AF (overinstrumentation) did not create dentinal microcracks in the roots of extracted 2-rooted maxillary premolars.