On the Causality Between Dentinal Defects and Root Canal Preparation: A Micro-CT Assessment.

This study aimed to evaluate the cause-effect relationship between canal preparation with ProTaper Universal (PTU) system and dentinal defects formation using micro-computed tomography (micro-CT) analysis. Forty mesial canals of mandibular molars with a type II Vertucci's canal configuration were scanned at an isotropic resolution of 14.16 µm. The sample was assigned to an experimental (n = 30) and a control (n = 10) groups, and the mesial canals were prepared with PTU system up to F2 instrument. The specimens from the experimental group were scanned and the cross-section images of the mesial roots, before and after preparation, were screened to identify the presence of dentinal defects. In the control group, the specimens were sectioned perpendicularly to the long axis of the root into 1-mm-thick slices (n = 80) and examined under optical microscope. Once a dentinal defect was detected, the slice was scanned through micro-CT. In the experimental group, dentinal micro-cracks were observed in 4,828 slices (24.04%). In all cross-section images, dentinal defects identified in the postoperative images were already present in the corresponding preoperative image. In the control group, 13 out of 80 slices (16.25%) had at least one dentinal defect visualized under stereomicroscopy, which was identified after a further micro-CT scanning. Micro-CT showed reliability as similar as optical microscopy in detecting dentinal defects, adding the possibility of tracking the dentinal tissue, before and after canal preparation, and providing a clear visualization of micro-cracks. Root canal preparation with PTU system did not induce the formation of new dentinal defects.

On the Causality Between Dentinal Defects and Root Canal Preparation: A Micro-CT Assessment

Abstract This study aimed to evaluate the cause-effect relationship between canal preparation with ProTaper Universal (PTU) system and dentinal defects formation using micro-computed tomography (micro-CT) analysis. Forty mesial canals of mandibular molars with a type II Vertucci's canal configuration were scanned at an isotropic resolution of 14.16 µm. The sample was assigned to an experimental (n = 30) and a control (n = 10) groups, and the mesial canals were prepared with PTU system up to F2 instrument. The specimens from the experimental group were scanned and the cross-section images of the mesial roots, before and after preparation, were screened to identify the presence of dentinal defects. In the control group, the specimens were sectioned perpendicularly to the long axis of the root into 1-mm-thick slices (n = 80) and examined under optical microscope. Once a dentinal defect was detected, the slice was scanned through micro-CT. In the experimental group, dentinal micro-cracks were observed in 4,828 slices (24.04%). In all cross-section images, dentinal defects identified in the postoperative images were already present in the corresponding preoperative image. In the control group, 13 out of 80 slices (16.25%) had at least one dentinal defect visualized under stereomicroscopy, which was identified after a further micro-CT scanning. Micro-CT showed reliability as similar as optical microscopy in detecting dentinal defects, adding the possibility of tracking the dentinal tissue, before and after canal preparation, and providing a clear visualization of micro-cracks. Root canal preparation with PTU system did not induce the formation of new dentinal defects.

Resumo O objetivo deste estudo foi avaliar a relação de causa-efeito entre o preparo do canal radicular com o sistema ProTaper Universal (PTU) e a formação de defeitos dentinários usando a análise por micro-tomografia computadorizada (micro-CT). Quarenta raízes mesiais de molares inferiores com a configuração tipo II de Vertucci foram escaneadas com resolução isotrópica de 14.16 μm. A amostra foi distribuída em um grupo experimental (n = 30) e um grupo controle (n = 10), e os canais mesiais foram preparados com o sistema PTU até a lima F2. As amostras do grupo experimental foram escaneadas e as imagens de secção transversal das raízes mesiais, antes e após o preparo, foram analisadas ​​para identificar a presença de defeitos dentinários. No grupo controle, os dentes foram seccionados perpendicularmente em relação ao longo eixo da raiz em fatias de 1 mm de espessura (n = 80) e examinados ao microscópio óptico. Uma vez detectado um defeito dentinário, a fatia foi escaneada através da micro-CT. No grupo experimental, defeitos dentinários foram observados em 4.828 secções transversais (24,04%). Em todas as imagens de secção transversal, os defeitos dentinários após os procedimentos experimentais já estavam presentes na secção pré-operatória correspondente. No grupo controle, 13 das 80 fatias (16,25%) apresentaram pelo menos um defeito dentinário visualizado através do microscópio, o qual foi identificado em um escaneamento posterior. A micro-CT mostrou confiabilidade com a microscopia óptica para detecção e visualização dos defeitos dentinários, permitindo acompanhar o tecido dentinário antes e depois do preparo do canal. O preparo do canal radicular com o sistema PTU não induziu a formação de novos defeitos dentinários.

Micro-CT Evaluation of Non-instrumented Canal Areas with Different Enlargements Performed by NiTi Systems

The aim of this study was to compare the percentage of non-instrumented area of root canals prepared with different enlargements using single-file reciprocating systems (Reciproc and WaveOne) and a conventional multi-file rotary (BioRaCe) system by micro-computed tomographic analysis. Thirty mesial roots of mandibular molars with moderate curvature (10° to 20°) presenting a type II Vertucci canal configuration and similar internal volume were chosen and scanned at an isotropic resolution of 14.16 µm. The sample was assigned to 3 groups (n=10) according to the system used for root canal preparation: Reciproc, WaveOne, and BioRaCe groups. Second and third scans were taken after the canals were prepared with instruments sizes 25 and 40, respectively. The recorded images of the surface area voxels of the canals, before and after preparation were examined from the furcation level to the apex to quantify the non-instrumented surface. Statistical data were compared using GLM for repeated-measures with a significance level set at 5%. Instrumentation systems did not influence the percentage of untouched root canal surfaces (p=0.690) whilst a significant reduction in the percentage of static voxels was observed after the enlargement of the root canal (p=0.010) in all groups (p=0.507). None of the systems was able to prepare the entire surface area of the mesial root canal of mandibular molars. The increased final apical size resulted in a significant positive effect on the shaping ability of the tested systems.

Resumo O objetivo deste estudo foi comparar a porcentagem de área não-instrumentada de canais radiculares preparados com diferentes ampliações utilizando sistemas reciprocantes de lima única (Reciproc e WaveOne) e um sistema rotatório convencional de múltiplas limas (BioRaCe) usando a análise da micro tomografia computadorizada. Trinta raízes mesiais de molares inferiores com curvatura moderada (10º a 20º) apresentando a configuração classe II de Vertucci e semelhança de volume do canal foram selecionadas e escaneadas em uma resolução isotrópica de 14,16 µm. A amostra foi dividida em 3 grupos (n=10) de acordo com o sistema utilizado para a preparação do canal radicular: grupo Reciproc, grupo WaveOne e grupo BioRaCe. Segundo e terceiro escaneamentos foram realizados após os canais serem preparados com instrumentos tamanhos 25 e 40, respectivamente. As imagens registradas de voxels da área de superfície dos canais, antes e após o preparo, foram examinadas desde o nível da furca até o ápice para quantificar a superfície não instrumentada. Os dados foram comparados estatisticamente através de GLM para medidas repetidas com um nível de significância de 5%. Os sistemas de instrumentação não influenciaram o percentual de superfícies não tocadas dos canais radiculares (p=0,690), enquanto que uma redução significativa no percentual de voxels estáticos foi observada após o alargamento do canal radicular (p=0,010) em todos os grupos (p=0,507). Nenhum dos sistemas foi capaz de preparar toda a área de superfície do canal mesial dos molares inferiores. O aumento do tamanho final apical resultou num efeito positivo significativo sobre a capacidade de modelar dos sistemas testados.

Accumulated hard tissue debris produced during reciprocating and rotary nickel-titanium canal preparation.

INTRODUCTION: This study compared the amount of hard tissue debris produced after different apical enlargement with single-file reciprocating systems (WaveOne [Dentsply Maillefer, Baillaigues, Switzerland] and Reciproc [VDW, Munich, Germany]) and a conventional multifile rotary system (BioRaCe [FKG Dentaire, La-Chaux-de-Fonds, Switzerland]) using micro-computed tomographic imaging. METHODS: Thirty moderately curved mesial roots of mandibular molars presenting 2 independent root canals were selected and scanned at an isotropic resolution of 14.16 µm. The sample was assigned to 3 groups (n = 10) with respect to the root length and degree of curvature of the mesial root according to the system used for the root canal preparation: Reciproc, WaveOne, and BioRaCe. Second and third scans were taken after the root canals were prepared up to ISO sizes 25 and 40, respectively. The matched images of the mesial canals, before and after preparation, were examined from the furcation level to the apex to evaluate the amount of hard tissue debris (%). Data were statistically compared using a general linear model for repeated-measures with a significance level set at 5%. RESULTS: Instrumentation systems per se did not influence the amount of hard tissue accumulation (P > .05), whereas a significant reduction in the percentage of hard tissue debris was observed after sequential enlargement in all groups (P < .05). CONCLUSIONS: None of the systems yielded root canals completely free from packed hard tissue debris. The increased final apical size resulted in significantly less debris accumulation for both reciprocating and rotary systems.

Micro-CT Evaluation of Non-instrumented Canal Areas with Different Enlargements Performed by NiTi Systems.

The aim of this study was to compare the percentage of non-instrumented area of root canals prepared with different enlargements using single-file reciprocating systems (Reciproc and WaveOne) and a conventional multi-file rotary (BioRaCe) system by micro-computed tomographic analysis. Thirty mesial roots of mandibular molars with moderate curvature (10° to 20°) presenting a type II Vertucci canal configuration and similar internal volume were chosen and scanned at an isotropic resolution of 14.16 µm. The sample was assigned to 3 groups (n=10) according to the system used for root canal preparation: Reciproc, WaveOne, and BioRaCe groups. Second and third scans were taken after the canals were prepared with instruments sizes 25 and 40, respectively. The recorded images of the surface area voxels of the canals, before and after preparation were examined from the furcation level to the apex to quantify the non-instrumented surface. Statistical data were compared using GLM for repeated-measures with a significance level set at 5%. Instrumentation systems did not influence the percentage of untouched root canal surfaces (p=0.690) whilst a significant reduction in the percentage of static voxels was observed after the enlargement of the root canal (p=0.010) in all groups (p=0.507). None of the systems was able to prepare the entire surface area of the mesial root canal of mandibular molars. The increased final apical size resulted in a significant positive effect on the shaping ability of the tested systems.

Lack of causal relationship between dentinal microcracks and root canal preparation with reciprocation systems.

INTRODUCTION: This study aimed to evaluate the frequency of dentinal microcracks observed after root canal preparation with 2 reciprocating and a conventional full-sequence rotary system using micro-computed tomographic analysis. METHODS: Thirty mesial roots of mandibular molars presenting a type II Vertucci canal configuration were scanned at an isotropic resolution of 14.16 µm. The sample was randomly assigned to 3 experimental groups (n = 10) according to the system used for the root canal preparation: group A-Reciproc (VDW, Munich, Germany), group B-WaveOne (Dentsply Maillefer, Baillagues, Switzerland), and group C-BioRaCe (FKG Dentaire, La-Chaux-de-Fonds, Switzerland). Second and third scans were taken after the root canals were prepared with instruments sizes 25 and 40, respectively. Then, pre- and postoperative cross-section images of the roots (N = 65,340) were screened to identify the presence of dentinal defects. RESULTS: Dentinal microcracks were observed in 8.72% (n = 5697), 11.01% (n = 7197), and 7.91% (n = 5169) of the cross-sections from groups A (Reciproc), B (WaveOne), and C (BioRaCe), respectively. All dentinal defects identified in the postoperative cross-sections were also observed in the corresponding preoperative images. CONCLUSIONS: No causal relationship between dentinal microcrack formation and canal preparation procedures with Reciproc, WaveOne, and BioRaCe systems was observed.

Assessing accumulated hard-tissue debris using micro-computed tomography and free software for image processing and analysis.

INTRODUCTION: The accumulation of debris occurs after root canal preparation procedures specifically in fins, isthmus, irregularities, and ramifications. The aim of this study was to present a step-by-step description of a new method used to longitudinally identify, measure, and 3-dimensionally map the accumulation of hard-tissue debris inside the root canal after biomechanical preparation using free software for image processing and analysis. METHODS: Three mandibular molars presenting the mesial root with a large isthmus width and a type II Vertucci's canal configuration were selected and scanned. The specimens were assigned to 1 of 3 experimental approaches: (1) 5.25% sodium hypochlorite + 17% EDTA, (2) bidistilled water, and (3) no irrigation. After root canal preparation, high-resolution scans of the teeth were accomplished, and free software packages were used to register and quantify the amount of accumulated hard-tissue debris in either canal space or isthmus areas. RESULTS: Canal preparation without irrigation resulted in 34.6% of its volume filled with hard-tissue debris, whereas the use of bidistilled water or NaOCl followed by EDTA showed a reduction in the percentage volume of debris to 16% and 11.3%, respectively. The closer the distance to the isthmus area was the larger the amount of accumulated debris regardless of the irrigating protocol used. CONCLUSIONS: Through the present method, it was possible to calculate the volume of hard-tissue debris in the isthmuses and in the root canal space. Free-software packages used for image reconstruction, registering, and analysis have shown to be promising for end-user application.

In vitro genotoxicity and cytotoxicity in murine fibroblasts exposed to EDTA, NaOCl, MTAD and citric acid

The aim of the present study was to evaluate the capacity of some root canal irrigants to induce genetic damage and/or cellular death in vitro. Murine fibroblast cells were exposed to ethylenediaminetetraacetic acid (EDTA), sodium hypochlorite (NaOCl), MTAD™ and citric acid in increasing concentrations for 3 h at 37ºC. The negative control group was treated with vehicle control (phosphate buffer solution - PBS) for 3 h at 37°C, and the positive control group was treated with methylmetanesulfonate, 1 μM. for 3 h at 37°C. Cytotoxicity was assessed by the trypan blue test and genotoxicity was evaluated by the single cell gel (comet) assay. The results showed that exposure to 2.5% and 5% NaOCl and 8.5% citric acid resulted in a significant cytotoxic effect. NaOCl, EDTA and citric acid did not produce genotoxic effects with respect to the comet assay data for all evaluated concentrations. Although MTAD was not a cytotoxic agent, it showed significant genotoxic effects at all tested concentrations (ANOVA and Tukey's test; p<0.05). NaOCl, EDTA and citric acid were found to be cytotoxic in a dose-dependent manner, but they were not genotoxic. MTAD did not cause cell death, but presented genotoxic effects.

O objetivo do presente estudo foi avaliar a capacidade de alguns irrigantes endodônticos em induzir danos genéticos e/ou morte celular in vitro. Células de fibroblastos murinos foram expostas ao ácido etilenodiaminotetracético (EDTA), hipoclorito de sódio (NaOCl), MTAD™ e ácido cítrico em concentrações crescentes durante 3 h a 37°C. O grupo controle negativo foi tratado com solução tampão fosfato - PBS por 3 h a 37° C e o grupo controle positivo foi tratado com metilmetanesulfonato a 1 μM por 3 h a 37° C. A citotoxicidade foi testada pelo azul de tripan e a genotoxicidade foi avaliada pelo teste do cometa. Os resultados apontaram que a exposição ao NaOCl a 2,5% e 5%, e ácido cítrico a 21% resultou em efeitos citotóxicos significativos. O NaOCl, EDTA e o ácido cítrico não produziram efeitos genotóxicos no que diz respeito aos dados obtidos pelo ensaio do Cometa em todas as concentrações testadas. Embora o MTAD não tenha sido um agente citotóxico, mostrou efeitos genotóxicos significativos em todas as concentrações testadas (ANOVA e teste de Tuckey; p<0,05). O NaOCl, o EDTA e o ácido cítrico mostraram-se citotóxicos de maneira dose-dependente, mas não genotóxicos. Por outro lado, apesar do MTAD não ter causado a morte celular, foi genotóxico em todas as concentrações testadas.

Mapping large extensions of flat dentin through digital microscopy: introduction to the method and possible applications.

PURPOSE: To introduce a mapping method to characterize large dentin surfaces using digital microscopy and to discuss the advantages and possible applications of the method. MATERIALS AND METHODS: Twenty unerupted third molars were sectioned transversally exposing coronal dentin surfaces. The microscopic mosaic method was used to generate a large field image with the resolution necessary to measure characteristics of dentin tubules. The AxioVision 4.7 software was used to control a motorized optical microscope and the process of acquiring approximately 400 small images to generate each dentin mosaic. An image analysis routine measured the number of tubules (NT) and the ratio between the total area of tubules and the area of the mosaic - the area fraction (AF) - of each mosaic. An automatic procedure transformed the mosaic image into a color map, providing a direct visual representation of tubule density through colors. The dentin maps were used for a comparative qualitative analysis of tubule density distribution of each sample. RESULTS: The results for NT (92450 to 196029 tubules/sample) and AF (4.12% to 11.10%) demonstrated a wide variation among dentin samples. The maps confirmed the microstructure variety, also revealing strong local variations in tubule density within each sample. CONCLUSION: The mapping method was able to perform dentin morphology characterization and is a valuable tool for producing a baseline for dentin adhesion studies. The method could be also useful in determining the real contribution of dentin structures to the final adhesion quality.

Self-adjusting file cleaning-shaping-irrigation system optimizes the filling of oval-shaped canals with thermoplasticized gutta-percha.

INTRODUCTION: This study aimed to compare the filling ability of carrier-based thermoplasticized gutta-percha in flat-oval canals prepared using either rotary ProTaper files (Dentsply Maillefer, Ballaigues, Switzerland) or the Self-Adjusting File system (SAF) (ReDent-Nova, Ra'anana, Israel). METHODS: Thirteen pairs of mandibular incisors were selected from a random collection. The teeth in each pair had single root canals with a flat-oval cross-section and similar sizes and dimensions. Teeth from each pair were randomly assigned to 1 of 2 experimental groups. One group was instrumented using the ProTaper NiTi system, whereas the SAF system was used in the other. Root filling was performed with Thermafil obturators (Dentsply Tulsa Dental Products, Tulsa, OK), and teeth were sectioned at 6, 5, 4, and 3 mm from the apex; the cut surface was subjected to morphometric measurement to establish the percent gutta-percha-filled area (PGFA) for each section. The Wilcoxon matched-pairs signed rank test was used to assess the effect of the 2 preparation methods on the PGFA. RESULTS: The median PGFA in the ProTaper group was 77.5%, whereas the median PGFA was 90.5% in the SAF group (P < .05). In the SAF-instrumented group, 17.8% of the specimens had a PGFA ≥95% compared with only 5.8% of the ProTaper-instrumented specimens (P < .05). CONCLUSIONS: Instrumentation of the flat-oval canals with the SAF system led to a significantly higher PGFA compared with ProTaper instrumentation with syringe and needle irrigation.

In vitro genotoxicity and cytotoxicity in murine fibroblasts exposed to EDTA, NaOCl, MTAD and citric acid.

The aim of the present study was to evaluate the capacity of some root canal irrigants to induce genetic damage and/or cellular death in vitro. Murine fibroblast cells were exposed to ethylenediaminetetraacetic acid (EDTA), sodium hypochlorite (NaOCl), MTAD™ and citric acid in increasing concentrations for 3 h at 37ºC. The negative control group was treated with vehicle control (phosphate buffer solution - PBS) for 3 h at 37°C, and the positive control group was treated with methylmetanesulfonate, 1 µM. for 3 h at 37°C. Cytotoxicity was assessed by the trypan blue test and genotoxicity was evaluated by the single cell gel (comet) assay. The results showed that exposure to 2.5% and 5% NaOCl and 8.5% citric acid resulted in a significant cytotoxic effect. NaOCl, EDTA and citric acid did not produce genotoxic effects with respect to the comet assay data for all evaluated concentrations. Although MTAD was not a cytotoxic agent, it showed significant genotoxic effects at all tested concentrations (ANOVA and Tukey's test; p<0.05). NaOCl, EDTA and citric acid were found to be cytotoxic in a dose-dependent manner, but they were not genotoxic. MTAD did not cause cell death, but presented genotoxic effects.

Biopure MTAD Induces DNA Damage but Not Cellular Death: An In Vitro Study.

OBJECTIVES: The purpose of this study was to evaluate the capacity of BioPure MTAD to induce genetic damage in vitro. Genotoxicity was assessed by the single cell gel (comet) assay. METHODS: Chinese hamster ovary (CHO) cells or murine fibroblasts cells were exposed to increasing final concentrations ranging from 0.1 a 10%. All treatments were performed for 1 hour at 37 degrees C. The negative control group was treated with vehicle control (phosphate buffer solution - PBS) for 1 hour at 37 degrees C and the positive control group was treated with methylmetanesulfonate (at 1 muM) for 1 hour at 37 degrees C. RESULTS: Present results showed that the BioPure MTAD was able to promote DNA breakage in CHO cells only at the highest concentration tested as well as to induce significant increase in tail moment at all tested concentrations in murine fibroblasts. CONCLUSIONS: In summary, our results indicate that BioPure MTAD is a genotoxic agent as depicted by the single cell gel (comet) assay. (Eur J Dent 2009;3:285-289).