Notch induces transcription by stimulating release of paused RNA Pol II without increasing chromatin accessibility.

Notch proteins undergo ligand-induced proteolysis to release a nuclear effector that influences a wide range of cellular processes by regulating transcription. Despite years of study, however, how Notch induces the transcription of its target genes remains unclear. Here, we comprehensively examined the response to human Notch1 across a time course of activation using genomic assays of nascent RNA and chromatin accessibility. These data revealed that Notch induces target gene transcription primarily by releasing paused RNA polymerase II (RNAPII), in contrast to prevailing models suggesting that Notch acts by promoting chromatin accessibility. Indeed, we found that open chromatin is established at Notch-responsive regulatory elements prior to Notch signaling, through SWI/SNF-mediated remodeling. Notch activation, however, elicited no further chromatin opening at these loci. Together, these studies reveal that the nuclear response to Notch signaling is dictated by the pre-existing chromatin state and RNAPII distribution at time of signal activation.

Effect of Autoclave Sterilization on the Number of Uses and Resistance to Cyclic Fatigue of WaveOne Gold and Four Replica-Like Endodontic Instruments.

This study aimed to evaluate the effect of autoclave sterilization on the integrity and instruments' fracture number after multiple uses and cyclic fatigue of the original WaveOne Gold (Dentsply Sirona Endodontics) compared to four replica-like instruments (TF4-Gold, Roll-Wave-Gold, W-File, and Micro-Gold). The instruments were analyzed by scanning electron microscope (SEM) before being used in root canal instrumentation (baseline). One hundred and fifty human molars, freshly extracted for orthodontic reasons or periodontal disease and with severe curvature (between 30° and 60°), were used. Fifty teeth were instrumented with 10 instruments from each group and were evaluated for integrity. After sterilization in an autoclave, the instruments were analyzed by SEM. This procedure was repeated twice more, totaling three rounds of instrumentation, sterilization, and SEM analysis. Ten unused instruments from each group were evaluated for resistance to cyclic fatigue in a static test using a motor and a device simulating a canal with a 60° curvature angle. The instruments were driven by the motor until separation, visually verified, and the time measured in seconds. Data were analyzed by Chi-square, one-way ANOVA, and Tukey analysis, considering a significance level of 5%. It was found that there was no statistically significant difference between the groups tested in the effect of sterilization on the number of uses. The SEM analysis showed distortions in the instruments after the 3rd use. There was a statistically significant difference in the cyclic fatigue test between the results of WaveOne Gold, TF4 Gold, and Roll Wave Gold compared to W File and Micro Gold (P < 0.0001) and a statistically significant difference between the W File and Micro Gold groups (P < 0.0001). In conclusion, this study affirmed that WaveOne Gold, TF4-Gold, and Roll-Wave-Gold instruments exhibit comparable cyclic fatigue resistance. Besides, all examined instruments can be reliably employed for up to two cases.

Analysis of morphology and symmetry of the root canal system of incisors, premolars and mandibular molars using CBCT.

Knowledge of root canal internal anatomy and its variations is important forproper endodontic treatment. It is therefore necessary to investigate morphological aspects among different dental groups in the same patient to define the best protocol for the case. AIM: To evaluate the morphology and symmetry of homologous incisors, premolars and mandibular molars using cone beam computed tomography (CBCT). MATERIALS AND METHOD: Descriptive statistical analysis was performed for the frequency of categorical variables, and a chi-square test or Fisher 's exact test was used to test whether gender and side were associated with number of roots, number of canals, and Vertucci's classification. Forty-five CBCT scans were evaluated, and 444 mandibular teeth were analyzed. The number of roots, number of canals, classification of the canals in each root according to Vertucci and presence of a symmetrical relationship between pairs of posterior teeth were analyzed. RESULTS: The resuls showed that 74% of mandibular central incisors had type I root canal, 26% of mandibular lateral incisors had type I and, with a significant difference in the number of canals between males and females (p < 0.05). In mandibular first premolars, 70.5% had type I; and in mandibular second premolars, 98.5% had type I. Mandibular first molars had two roots in 98% of the cases. Second mandibular molars had two roots in 92.5% of the cases, one root in 6%, and three roots in 1.5%. Symmetry between central incisors was higher in females than in males. CONCLUSIÓN: Teeth of the same group can have different morphologies in the same patient.

0 conhecimento da anatomia interna e suas variagoes anatómicas é fator importante para o adequado tratamento endodóntico. Portanto, é necessário investigar esses aspectos morfológicos entre diferentes grupos dentários de um mesmo paciente para definir o melhor protocolo para o caso. OBJETIVO: Avaliar a morfologia e simetria de incisivos, pré-molares e molares inferiores homólogos por meio de tomografia computadorizada de feixe cónico (TCFC). MATERIAIS E MÉTODO: Foi realizada análise estatística descri-tiva para a frequéncia das variáveis categóricas e foi utilizado o teste do qui-quadrado ou teste exato de Fisher para testar a relagao entre sexo e lado em comparagao com número de raízes, número de canais e classificagao de Vertucci. Quarenta e cinco TCFC foram avaliadas e 444 dentes inferiores foram analisados. Foram considerados: o número de raízes, o número de canais, o tipo dos canais acordo com a classificagao de Vertucci e a presenga de relagao simétrica entre pares de dentes posteriores. RESULTADOS: Os resultados mostraram que 74% dos incisivos centrais inferiores tinham um canal radicular tipo 1 e 26% tinham dois canais; 73% dos incisivos laterais inferiores, 26%oeram do tipo I, tinham um canal e 27% tinham dois canais, com diferenga significativa no número de canais entre os grupos masculino e feminino (p < 0,05). Nos primeiros pré-molares inferiores, tipo I, um canal foi detectado em 70,5% e dois canais em 29,5%; nos segundos pré-molares inferiores, tipo I, um único canal foi detectado em 98,5%. O primeiro molar inferior foi observado com duas raízes em 98% e tres raízes em 2%o. O segundo molar inferior tinha duas raízes em 92,5% dos casos, uma raiz em 6% e tres raízes em 1,5%. A simetria foi maior nas mulheres em comparagao aos homens nos incisivos centrais. CONCLUSÃO: Pode-se concluir que dentes de um mesmo grupo podem apresentar morfologias diferentes no mesmo paciente.

Relationship between periapical lesions and sinus changes on multi-slice computed tomography sean.

Differentiating orofacial odontogenic pain/disorders from pain/disorders associated with maxillary sinusitis is important to avoid unnecessary dentalprocedures and to properly refer patients to colleagues/dentists and vice versa. AIM: To analyze the association between apical lesions and sinus changes and to evaluate the agreement between the diagnoses of an endodontist, a radiologist, an oral and maxillofacial surgeon, and an otolaryngologist. MATERIALS AND METHOD: 385 axial, coronal, and sagittal MSCT scans were selected using an image archiving andcommunication system (PACS). The examinations had been performed between 2018 and 2022. RESULTS: Apical lesions were observed in 36.10% of sinusitis cases, 73.8% of unilateralsinusitis cases, 48.7% of sinus floor discontinuity cases, and 67.2% of cases in which endodontic treatment had been performed. Agreement between the diagnoses made by the endodontist and those made by the other investigators was high for most study variables (k > 0.60). The exceptions were mucosal thickening, for which agreement between the endodontist and the other investigators was intermediate (k=0.397), and the presence of periapicallesions (k=0.010), previous endodontic treatment (k=0.013), and mucosal thickness (k=0.024), for which agreement between endodontists and radiologists was low. Conclusions: There was an association between sinus changes and apical lesions.

Diferenciar a dor/desordens odontogénicas orofaciais da dor/desordens as sociadas á sinusite maxilar é importante para evitar procedimentos odontológicos desnecessários e para encaminhar adequadamente os pacientes aos colegas/dentistas e vice-versa. OBJETIVO: Analisar a associagdo entre lesoes apicais e alteragóes sinusais e avaliar a concordáncia entre os diagnósticos de um endodontista, um radiologista, um cirurgido bucomaxilofacial e um otorrinolaringologista. MATERIAL E MÉTODO: foram avaliadas 385 imagens. RESULTADOS: Aslesoes apicais foram observadas em 36,10% dos casos de sinusite, em 73,8% dos casos de sinusite unilateral, em 48,7% dos casos de descontinuidade do assoalho do seio e em 67,2%odos casos em que o tratamento endodontico havia sido realizado. A concordancia entre os diagnósticos feitos pelo endodontista e os feitos pelos outros pesquisadores foi alta para a maioria das variáveis do estudo (k > 0,60). As excegoes foram o espessamento da mucosa, para o qual a concordáncia entre o endodontista e os outros pesquisadores foi intermediária (k=0,397) e a presenga delesoes periapicais (k=0,010), tratamento endodontico prévio (k=0,013) e espessura da mucosa (k=0,024), para os quais a concordáncia entre endodontistas e radiologistas foi baixa. CONCLUSÕES: Houve uma associagdo entre as alteragóes sinusais e aslesoes apicais.

In vitro study of structural and chemical changes in two reciprocating files after multiple reuses and sterilisation.

To use scanning electron microscopy and energy dispersive X-ray spectroscopy to evaluate the metallurgical-chemical changes of WaveOne Gold (WOG) and R-Motion (RM), after multiple uses. The instruments were divided into groups (n = 8): WOG and RM-control groups, new instruments; WOG1 and RM1; WOG2 and RM2; WOG3 and RM3 after instrumentation of 1, 2 or 3 molars, respectively. Burrs occurred mainly in the control group and after the first use. The RM files were found to have a higher nickel content, which increased during reuse, and a decrease in oxygen content with increasing reuse, in addition to calcium impregnation, which occurred in greater amounts in the corrosion areas in the WOG files. The presence of topographic and chemical changes was demonstrated, indicating that caution should be taken when reusing endodontic instruments to avoid fractures.

Temporal dynamics and stoichiometry in human Notch signaling from Notch synaptic complex formation to nuclear entry of the Notch intracellular domain.

Mammalian Notch signaling occurs when the binding of Delta or Jagged to Notch stimulates the proteolytic release of the Notch intracellular domain (NICD), which enters the nucleus to control target gene expression. To determine the temporal dynamics of events associated with Notch signaling under native conditions, we fluorescently tagged Notch and Delta at their endogenous genomic loci and visualized them upon pairing of receiver (Notch) and sender (Delta) cells as a function of time after cell contact. At contact sites, Notch and Delta immediately accumulated at 1:1 stoichiometry in synapses, which resolved by 15-20 min after contact. Synapse formation preceded the entrance of the Notch extracellular domain into the sender cell and accumulation of NICD in the nucleus of the receiver cell, which approached a maximum after ∼45 min and was prevented by chemical and genetic inhibitors of signaling. These findings directly link Notch-Delta synapse dynamics to NICD production with spatiotemporal precision.

Influence of Passive Ultrasonic Irrigation of the Photosensitizer Used in Photodynamic Therapy on Microbial Reduction in the Root Canal System: An in Vitro Study.

Introduction: The success of endodontic treatment is based on microbial reduction promoted by the interaction of chemical and mechanical procedures. Photodynamic therapy (PDT) is used as an adjunct to conventional treatment, with significant microbial reduction. This study aimed to evaluate the influence of passive ultrasonic irrigation (PUI) of the photosensitizer (PS) used in PDT on microbial reduction in the root canal system. Materials and Methods: Forty-five mesiobuccal root canals from extracted human mandibular molars that were inoculated with standard strains of Enterococcus faecalis, Candida albicans, and Streptococcus mutans for 21 days were selected. The root canals were prepared using the ProTaper Next system and randomly divided into 3 groups (n=15): 1) PDT: PDT alone; 2) PUI+PDT: PUI followed by PDT; and 3) PUI/PS+PDT: PUI of the PS followed by PDT. Microbiological samples were collected from the canals before and after using the protocols described above. The data were analyzed by analysis of variance (Tukey's test) at a significance level of 5%. Results: Microbial counts before PDT, PUI+PDT, and PUI/PS+PDT did not differ significantly (P>0.05), showing methodological standardization in the microbial contamination of the root canal system. There was a significant reduction in microbial counts after PDT (61.05%), PUI+PDT (65.04%), and PUI/PS+PDT (68.58%) (P<0.01), but with no statistically significant difference between the three groups (P>0.05). Conclusion: Based on the present findings, it was concluded that the combination of PUI with PDT or PS activation did not influence microbial reduction achieved by PDT alone.

Comparative in vitro analysis of the antifungal activity of different calcium silicate-based endodontic sealers

Aim: This study aimed to perform an in vitro comparative analysis of the antifungal activity of different calcium silicate-based endodontic sealers against three fungal species. Methods: The antifungal properties of three calcium silicate-based sealers were tested: Bio-C Sealer, Cambiar a Sealer Plus BC, and MTA-Fillapex. Two commonly used sealers were used as controls: AH Plus and Endomethasone. An agar diffusion test was performed to analyze the antifungal activity of the sealers against Candida albicans, Candida glabrata, Candida tropicalis, and a mixed microbial culture medium. The results were analyzed using ANOVA (p <0.05). Results: Endomethasone exhibited the highest inhibition against all strains examined, maintaining a consistent level of inhibition throughout 7 days. MTA-Fillapex demonstrated the best performance among the calcium silicate-based sealers for the three fungal species (p < 0.05), maintaining stable values over the 7 days, surpassing that of Endomethasone. Nevertheless, MTA-Fillapex only exhibited antimicrobial effect against the mixed culture for the first 24 hours, and no antimicrobial activity was observed at 48 hours, being surpassed by all tested sealers (p < 0.05). Conclusion: Of all silicate-based sealers tested, only MTA-Fillapex exhibited promising antifungal activity. Nevertheless, care must be taken when extrapolating these results, as MTA-Fillapex exhibited poor antimicrobial activity when tested in mixed microbial cultures

Evaluation of the Ability of 3 Reciprocating Instruments to Remove Obturation Material: A Micro-Computed Tomography Study.

INTRODUCTION: The purpose of this study was to evaluate the ability of the Reciproc, Reciproc Blue, and WaveOne Gold systems to remove filling material during endodontic retreatment of extracted human mandibular premolars. METHODS: Thirty-nine teeth were instrumented with the Protaper Universal System to the F3 file and filled with the Tagger hybrid technique using an F3 gutta-percha cone and AH Plus cement. At the end of this period, the teeth were scanned with micro-computed tomography before and after removal of the filling material from the root canals. The teeth were divided into 3 groups (n = 13) based on the apical volume, depending on the systems used to remove the filling material. Group GR: Reciproc 40/.06; Group GRB: Reciproc Blue 40/.06; and Group GWG: WaveOne Gold 35/.06. The results were statistically analyzed using the tests of Kruskal-Wallis, Duncan, and analysis of variance at a significance level of 5%. RESULTS: The results showed that there were no significant differences between the amounts of filling material removed, either for the apical and middle regions alone or in the overall evaluation for the 3 groups (P = .97). The time evaluation statistically showed that the GR and GWG groups required less time to clean the root canals than the GRB group. CONCLUSIONS: Reciproc R40 files and WaveOne Gold Medium files required less time for endodontic treatment than Reciproc Blue R40 files. There was no difference in the ability to remove obturation material between the 3 instruments. No instrument was able to completely remove the filling material from the root canals.

Effect of obturation technique on dentinal tubule penetration of two tricalcium silicate-based sealers: Ex vivo study.

The aim of this study was to assess by confocal laser microscope the depth of dentinal tubule penetration of two tricalcium silicate-based sealers promoted by two obturation techniques in curved canals compared with AHPlus. One hundred and twenty canals were divided into six groups (n = 20): BCSC-Bio-C Sealer (BC) and single-cone technique (SC); BCCW-BC and continuous condensation wave (CW); TFSC-Total Fill (TF) and SC; TFCW-TF and CW; AHSC-AH Plus (AH) and SC; AHCW-AH and CW. Data were analysed using the three-way ANOVA and Tukey's test (α = 5%). Penetration depth was significantly greater for TFCW than TFSC and greater for AHCW than AHSC (p < 0.05). There was no significant difference between BCCW and BCSC (p > 0.05). The penetration of TF was significantly greater (p < 0.05). The CW technique promoted greater intratubular penetration, except for the BC sealer.

Assessing the cyclic fatigue resistance and sterilization effects on replica-like endodontic instruments compared to Reciproc Blue.

This study aimed to evaluate the effect of the number of uses and autoclave sterilization on the cyclic fatigue resistance of four replica-like instruments RC Blue; Only One File Blue; Recip One Blue; and Micro Blue compared to the original system Reciproc Blue (VDW, Munich, Germany). The instruments were analyzed by scanning electron microscope (SEM) before being used in root canal instrumentation (baseline). Fifty molars were instrumented by ten instruments (n=10). After sterilization in an autoclave, the  instruments were analyzed by SEM. This procedure was repeated twice more using different molars, totaling 3 rounds of instrumentation, sterilization and SEM analysis. Then, ten different instruments from each brand were tested for cyclic fatigue resistance. Number of uses data were analyzed using Chi-squared analysis, and cyclic fatigue data were analyzed by one-way Anova followed by Tukey's test as the data had normal distribution. The fracture times for all systems had no significant difference, but Micro Blue had significantly lower values than the other systems (p < 0.05). The SEM analysis showed distortions in the instruments after the 3rd use. Therefore, all tested instruments except of Micro Blue have similar resistance to cyclic fatigue, and all are reliable for use in up to 2-cases.

Temporal Dynamics and Stoichiometry in Notch Signaling - from Notch Synaptic Complex Formation to NICD Nuclear Entry.

Mammalian Notch signaling occurs when binding of Delta or Jagged to Notch stimulates proteolytic release of the Notch intracellular domain (NICD), which enters the nucleus to regulate target gene expression. To determine the temporal dynamics of events associated with Notch signaling under native conditions, we fluorescently tagged Notch and Delta at their endogenous genomic loci and visualized them upon pairing of receiver (Notch) and sender (Delta) cells as a function of time after cell contact. At contact sites, Notch and Delta immediately accumulated at 1:1 stoichiometry in synapses, which resolved by 15-20 min after contact. Synapse formation preceded entrance of the Notch extracellular domain into the sender cell and accumulation of NICD in the nucleus of the receiver cell, which approached a maximum after ∼45 min and was prevented by chemical and genetic inhibitors of signaling. These findings directly link Notch-Delta synapse dynamics to NICD production with unprecedented spatiotemporal precision.

A spatiotemporal Notch interaction map from plasma membrane to nucleus.

Notch signaling relies on ligand-induced proteolysis of the transmembrane receptor Notch to liberate a nuclear effector that drives cell fate decisions. Upon ligand binding, sequential cleavage of Notch by the transmembrane protease ADAM10 and the intracellular protease γ-secretase releases the Notch intracellular domain (NICD), which translocates to the nucleus and forms a complex that induces target gene transcription. To map the location and timing of the individual steps required for the proteolysis and movement of Notch from the plasma membrane to the nucleus, we used proximity labeling with quantitative, multiplexed mass spectrometry to monitor the interaction partners of endogenous NOTCH2 after ligand stimulation in the presence of a γ-secretase inhibitor and as a function of time after inhibitor removal. Our studies showed that γ-secretase-mediated cleavage of NOTCH2 occurred in an intracellular compartment and that formation of nuclear complexes and recruitment of chromatin-modifying enzymes occurred within 45 min of inhibitor washout. These findings provide a detailed spatiotemporal map tracking the path of Notch from the plasma membrane to the nucleus and identify signaling events that are potential targets for modulating Notch activity.

Analysis of antimicrobial efficacy of sodium hypochlorite and ozonated water against biofilm in oval canals.

This in vitro study compared the antimicrobial efficacy of 2.5% sodium hypochlorite (NaOCl) and 8 µg/mL ozonated water agitated by passive ultrasonic irrigation (PUI) or PUI combined with EndoActivator (EA) against mature multispecies biofilm. One hundred and five oval-shaped mandibular premolars were instrumented, sterilized, and inoculated with Enterococcus faecalis, Candida albicans, and Staphylococcus aureus, divided into: control group - saline; O3 group - ozonated water; O3 PUI group - ozonated water with PUI agitation; O3 PUI+EA group - ozonated water with PUI+EA agitation; NaOCl group - NaOCl; NaOCl PUI group - NaOCl with PUI agitation; and NaOCl PUI+EA group - NaOCl with PUI+EA agitation. Microbiological samples were collected before (S1) and after (S2) the disinfection procedures and the data were statistically analyzed using the Kruskal-Wallis test. In the culture method, there was significant disinfection in the O3 PUI+EA, NaOCl, NaOCl PUI, and NaOCl PUI+EA groups (p˂0.05). The combination of NaOCl with PUI+EA reduced microbial counts to zero (p˂0.05). In the qPCR method, there was a significant reduction in the total count of viable microorganisms in the O3 PUI, O3 PUI+EA, NaOCl, NaOCl PUI, and NaOCl PUI+EA groups (p˂0.05). It can be concluded that 2.5% NaOCl with and without agitation, as well as 8 µg/mL ozonated water with its action enhanced by the agitation techniques, were effective in root canal disinfection, and their antimicrobial efficacy is related to the microorganisms present in the biofilm.

Temporo-basal sulcal connections: a manual annotation protocol and an investigation of sexual dimorphism and heritability.

The temporo-basal region of the human brain is composed of the collateral, the occipito-temporal, and the rhinal sulci. We manually rated (using a novel protocol) the connections between rhinal/collateral (RS-CS), collateral/occipito-temporal (CS-OTS) and rhinal/occipito-temporal (RS-OTS) sulci, using the MRI of nearly 3400 individuals including around 1000 twins. We reported both the associations between sulcal polymorphisms as well with a wide range of demographics (e.g. age, sex, handedness). Finally, we also estimated the heritability, and the genetic correlation between sulcal connections. We reported the frequency of the sulcal connections in the general population, which were hemisphere dependent. We found a sexual dimorphism of the connections, especially marked in the right hemisphere, with a CS-OTS connection more frequent in females (approximately 35-40% versus 20-25% in males) and an RS-CS connection more common in males (approximately 40-45% versus 25-30% in females). We confirmed associations between sulcal connections and characteristics of incomplete hippocampal inversion (IHI). We estimated the broad sense heritability to be 0.28-0.45 for RS-CS and CS-OTS connections, with hints of dominant contribution for the RS-CS connection. The connections appeared to share some of their genetic causing factors as indicated by strong genetic correlations. Heritability appeared much smaller for the (rarer) RS-OTS connection.

Analysis of antimicrobial efficacy of sodium hypochlorite and ozonated water against biofilm in oval canals

Abstract This in vitro study compared the antimicrobial efficacy of 2.5% sodium hypochlorite (NaOCl) and 8 µg/mL ozonated water agitated by passive ultrasonic irrigation (PUI) or PUI combined with EndoActivator (EA) against mature multispecies biofilm. One hundred and five oval-shaped mandibular premolars were instrumented, sterilized, and inoculated with Enterococcus faecalis, Candida albicans, and Staphylococcus aureus, divided into: control group - saline; O3 group - ozonated water; O3 PUI group - ozonated water with PUI agitation; O3 PUI+EA group - ozonated water with PUI+EA agitation; NaOCl group - NaOCl; NaOCl PUI group - NaOCl with PUI agitation; and NaOCl PUI+EA group - NaOCl with PUI+EA agitation. Microbiological samples were collected before (S1) and after (S2) the disinfection procedures and the data were statistically analyzed using the Kruskal-Wallis test. In the culture method, there was significant disinfection in the O3 PUI+EA, NaOCl, NaOCl PUI, and NaOCl PUI+EA groups (p˂0.05). The combination of NaOCl with PUI+EA reduced microbial counts to zero (p˂0.05). In the qPCR method, there was a significant reduction in the total count of viable microorganisms in the O3 PUI, O3 PUI+EA, NaOCl, NaOCl PUI, and NaOCl PUI+EA groups (p˂0.05). It can be concluded that 2.5% NaOCl with and without agitation, as well as 8 µg/mL ozonated water with its action enhanced by the agitation techniques, were effective in root canal disinfection, and their antimicrobial efficacy is related to the microorganisms present in the biofilm.

Resumo Este estudo in vitro comparou a desinfecção do hipoclorito de sódio 2,5% (NaOCl) e da água ozonizada 8 µg/mL agitados pela irrigação ultrassônica passiva (PUI) e por associação da PUI com EndoActivator (EA) na redução de biofilme misto maduro. Cento e cinco pré-molares inferiores ovalados foram instrumentados, esterilizados e inoculados com Enterococcus faecalis, Candida albicans e Staphylococcus aureus, divididos em: Grupo controle: soro; Grupo O3: água ozonizada; Grupo O3 PUI: água ozonizada agitada por PUI; Grupo O3 PUI + EA: água ozonizada agitada por PUI e EA: Grupo NaOCl: hipoclorito de sódio; Grupo NaOCl PUI: hipoclorito de sódio agitado por PUI; Grupo NaOCl PUI + EA: hipoclorito de sódio agitado por PUI e EA. Amostras microbiológicas foram coletadas antes (S1) e após (S2) os procedimentos de desinfecção e os dados foram analisados estatisticamente pelo teste de Kruskal-Wallis. No método de cultura, houve desinfecção significativa nos grupos O3 PUI + EA, NaOCl, NaOCl PUI e NaOCl PUI + EA (p˂0.05), sendo que no grupo NaOCl PUI + EA não houve crescimento de microrganismo (p˂0.05). No método de qPCR, nas contagens dos microrganismos antes e após os protocolos de desinfecção, houve redução microbiana nos grupos O3 PUI, O3 PUI + EA, NaOCl, NaOCl PUI, NaOCl PUI + EA (p˂0.05). Concluiu-se que o NaOCl 2,5% com e sem agitação foi eficiente, assim como a água ozonizada 8 µg/mL potencializada pelos métodos de agitação na desinfecção e que a mesma está relacionada com os microrganismos presentes no biofilme.

Ex Vivo Evaluation of the Efficacy of Photodynamic Therapy in Eliminating Enterococcus faecalis from Dentinal Tubules by Confocal Laser Scanning Microscopy.

INTRODUCTION: The aim of the present study was to investigate ex vivo by confocal laser scanning microscopy the antibacterial effect of photodynamic therapy (PDT) on dentinal tubules in the apical 5 mm of human mandibular premolars contaminated with Enterococcus faecalis. METHODS: Thirty-four teeth were standardized to 20 mm and foraminal anatomic diameters using a #20 K-file (Dentsply Maillefer). Samples were contaminated for 21 days and divided into the following 3 experimental groups (n = 10): the PDT group (instrumented canals and PDT), the passive ultrasonic irrigation (PUI) group (instrumented canals and PUI), and the PUI-PDT group (instrumented canals, PUI, and PDT), along with a control group (n = 4) (noninstrumented canals). The canals in the experimental groups were instrumented with ProTaper Next (Dentsply Maillefer) up to X3 and rinsed with EDTA and sodium hypochlorite. The photosensitizer used was 0.01% methylene blue with a preirradiation time of 5 minutes and a diode laser with 4 J energy and a 660-nm wavelength. Cross sections were made 5 mm from the apex of all samples, which were analyzed using confocal laser scanning microscopy. The results were analyzed using the Shapiro-Wilk and Kruskal-Wallis (Dunn) tests. RESULTS: There was a lower percentage of live bacteria in the PUI-PDT group, with a statistical difference compared with the control and PDT groups (P < .05). There was no statistical difference in the percentage of live bacteria between PUI-PDT and PUI (P > .05). CONCLUSIONS: It was concluded that the PUI-PDT association was most effective in disinfecting root canals compared with the control group and PDT.

Micro- CT evaluation of sealers removal by reciprocal instrumentation followed by continuous ultrasonic irrigation in teeth with oval root canals.

Background: The ability of the Reciproc system (R40) followed by continuous ultrasonic irrigation (CUI) to remove filling material from oval root canals of mandibular premolars filled with gutta-percha and AH Plus or Bio-C Sealer was evaluated by microtomography (micro-CT). Material and Methods: The straight and oval root canals of 42 mandibular premolars were prepared with the ProDesign R 35.05 reciprocal file and randomly divided into two groups according to the material used to fill the canals (n=21): Group AH - Master Cone and AH Plus; Group BC - Master Cone and Bio-C Sealer. After filling and provisional sealing, the teeth were stored at 100% relative humidity and a temperature of 37°C for 30 days. The filling material was then removed with an R40 file. The material was considered completely removed when the R40 file reached the working length (WL), and no remaining filling material was visible on the canal walls. CUI was then performed. The teeth were scanned by micro- CT before and after removal of the filling material. The remaining filling material was measured in mm in the last apical 5 mm. The data were analyzed with the nonparametric Friedman test and subsequently with the Dunn test. The Mann-Whitney U test was also performed. Statistical significance was accepted at the 5% level. Results: After instrumentation with the Reciproc R40, the volume of residual filling material was significantly greater in the BC group than in the AH group (P = 0.035). After CUI, there was no difference in the volume of residual material between the two groups (P = 0.705). Conclusions: Bio-C sealer was more difficult to remove with the Reciproc file than AH Plus. CUI improved the removal of residual filling material regardless of sealer type. However, no technique was able to completely clear the canals of filling material. Key words:Bioceramic cement, CUI, micro-CT, reciproc, retreatment.

Microtomographic Assessment of the Shaping Ability of the Hyflex CM and XP-endo Shaper Systems in Curved Root Canals.

OBJECTIVE: This study compared the shaping ability of the Hyflex CM and XP-endo Shaper rotary file systems in curved mesial canals of mandibular molars using micro-computed tomography. MATERIAL AND METHODS: Seventeen mesial roots of extracted first mandibular molars with two independent mesial canals were scanned before and after root canal preparation with the tested rotatory file systems. Each mesial canal from the same specimen was prepared with one of the two systems. The parameters analyzed were canal centering (transportation) for the cervical, middle, and apical segments, as well as for the entire canal (0-10 mm from the apex); and canal volume increase, canal surface area increase, and unprepared canal walls for two segments, 0 to 4 mm and 0 to 10 mm from the apex. RESULTS: There was no significant difference between both systems regarding canal centering (transportation), volume increase, and unprepared canal walls for the 0 to 10 mm segment (p> 0.05); however, a significant difference was observed for the 0 to 4 mm segment (p <0.01), where the Hyflex CM left 28.46% of unprepared walls and XP-endo Shaper left 13.26%. CONCLUSIONS: The shaping ability of the two tested rotatory file systems in mesial roots of first mandibular molars was similar for all parameters in all the segments evaluated, except for the 0 to 4 mm segment, where XP-endo Shaper left a smaller area of unprepared canal walls than Hyflex CM.

Effect of different activation methods on the intratubular penetration of CaOH2 paste: Ex-vivo analysis by confocal laser scanning microscopy.

The aim of this study was to evaluate the maximum penetration depth (MPD) and penetration area (PA) of CaOH2 paste agitated by different methods and to measure the pH after performing these methods. Fifty-five mandibular premolars were divided into control, GL and 4 experimental groups, GEC, GXP, GEDDY and GI (n = 11), where the paste was agitated with Easy Clean, XP-endo Finisher, EDDY and Irrisonic, respectively. The canals were instrumented and irrigated with NaOCl. The paste was manipulated with propylene glycol and rhodamine B dye. Root sections were examined using confocal laser scanning microscopy. There was no difference between groups regarding PA (p > 0.05). At 5 mm, MPD was greater in GEC than in GL or GEDDY, and greater in GI than in GL (p < 0.05). At 2 mm, MPD was greater in GEC than in the other groups (p < 0.05). In conclusion, both Easy Clean and Irrisonic promoted deeper paste penetration and were associated with higher pH levels.