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Objectives: To test and validate a new protocol for in vitro contamination of dentinal tubules using Fusobacterium nucleatum (F. nucleatum) by confocal laser scanning microscopy (CLSM), in addition to evaluating the effectiveness of conventional endodontic irrigants such as sodium hypochlorite (NaOCl) and chlorhexidine (CLX) on this biofilm. Material and methods: Thirty lower premolars were contaminated with F. nucleatum (ATCC 51190) for 7 days under anaerobic conditions using the proposed new model. The specimens were divided into a control group and experimental groups, according to the irrigants: NaOCl 2.5% and CLX 2%. Then, the samples were submitted for analysis by CLSM and the LIVE/DEAD technique to quantify bacterial viability. Data normality was verified by the Shapiro-Wilk test. Intragroup and intergroup comparisons were performed using the Kruskal-Wallis test, followed by Dunn's post-test. Results: The CLSM images obtained demonstrated the effectiveness of the proposed new contamination protocol, with a high percentage of viable bacteria in relation to the treated groups (p < 0.05). Lower viability values were observed for the 2.5% NaOCl group. Conclusion: The new contamination protocol resulted in a high and homogeneous percentage of viable bacteria in the dentinal tubules in all specimens evaluated. Both irrigating solutions proved to be effective in reducing the intratubular microbiota, especially 2.5% NaOCl.
ABSTRACT
The aim of this study was to evaluate the influence of surface topography of gutta-percha (GP) cones and plasticized disks of GP on the initial adhesion of Enterococcus faecalis (E. faecalis). The GP cones (Tanari and Dentsply brands) were cut 3 mm from the apical portion and fixed on a glass slide. To make the disks, the cones were thermoplasticized in standardized molds. The specimens were divided into groups according to the shape of the GP and the presence or absence of the bacteria. For contamination, the strain of E. faecalis (ATCC 29212) was used. The surface topography was analyzed using an atomic force microscope (AFM). The surface, roughness, and waviness parameters were evaluated by the Kruskal-Wallis and Dunn test. The comparison between disks and cones showed significant differences, where the cones were rougher, with a higher value attributed to the Dentsply cone (DC group). The same was observed for the waviness. After contamination, there was greater bacterial accumulation in cones, especially in their valleys, but both the surface and the topography became more homogeneous and smoother, with no differences between disks and cones of both brands. The topographic surface of the GP, at the micro and nanoscale, influences the initial adhesion of E. faecalis, with a greater tendency for contamination in regions associated with the presence of roughness and waviness. In this context, plasticization of GP is indicated, as it reduces surface irregularities compared to cones, contributing to less retention of bacteria.
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Aim: This study investigated the influence in vitro of different sodium hypochlorite (NaOCl) agitation protocols associated or not with DualRinse (HEDP) on the temperature of the solution. Methods: Forty-eight premolars were instrumented and their apical third sealed to allow a closed irrigation system. The teeth remained immersed in a basin of warm water (37°C). The teeth were divided into the groups: G1 (NaOCl+Passive Ultrasonic Irrigation (PUI)), G2 (NaOCl/HEDP + PUI), G3 (NaOCl + EasyClean (EC)) and G4 (NaOCl/HEDP + EC). The canals were filled with the respective solutions and after 180 seconds the first temperature measurement was taken (T0). Then, the solutions were agitated, following the different protocols, for 60 seconds and a new measurement was performed (T60). The temperature was measured using a digital thermometer for type "K" sensors that was inserted into the middle third of the teeth. At the end of the measurements, the teeth were sectioned and prepared for scanning electron microscopy. The dentinal wall of middle third was graded according to the amount of debris and smear layer remaining on the walls. The results were analyzed using ANOVA test and Tukey's multiple comparisons (p<0.05). Results: G1 and G2 had an average increase in temperature of 1.1°C and 1.65°C, respectively (p>0.05). EasyClean caused a decrease in the temperature of the solutions in both groups, without a significant statistical difference with T0 (p>0.05). Regarding cleaning, it was only possible to observe clean dentinal tubules in the groups with the chelator. PUI discretely increased the temperature of the solution, regardless of the solution. The opposite effect was observed after activation with EasyClean. Conclusion: The association of NaOCl with a chelating agent promoted the cleaning of the dentinal tubules
Subject(s)
Root Canal Irrigants , Sodium Hypochlorite , Temperature , Ultrasonics , Microscopy, Electron, Scanning , Chelating AgentsABSTRACT
The objective of the present study was to investigate the pH, volumetric alteration, antimicrobial action, and effect on biofilm matrix polysaccharides of calcium hydroxide (CH) pastes with different vehicles available in endodontics: CH + propylene glycol (CHP), UltraCal XS®, Metapaste®, and Metapex®. The pH was analyzed at different time intervals using a pH meter. For volumetric alteration, a microtomographic assay was performed before and after immersion in water. Enterococcus faecalis was chosen for microbiological tests. The bacterial viability and extracellular matrix were quantified with direct contact evaluation (dentin blocks) and at the intratubular level (dentin cylinders) using LIVE/DEAD BacLight and Calcofluor White dyes via confocal laser scanning microscopy (CLSM). Kruskal-Wallis and Dunn's tests were used to analyze pH and direct contact assays, while one-way ANOVA and Tukey tests were used to analyze volumetric alteration and intratubular decontamination (α = 0.05). Higher pH values were obtained during the initial days. Volumetric alterations were similar in all groups. Lower bacterial viability was obtained for dentin blocks and cylinders when CH pastes were used. UltraCal XS and Metapex had lower values for the extracellular matrix. The pH of all CH pastes decreased with time and did not promote medium alkalization for up to 30 days. CH paste can reduce bacterial viability through direct contact and at an intratubular level; however, UltraCal XS and Metapex are involved with lower volumes of extracellular matrices.
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BACKGROUND: There has been no unanimity concerning the ideal concentration of citric acid for safe use in clinical practice. This study evaluated the cytotoxicity and the antibacterial activity in infected dentinal tubules of 10% and 1% citric acid (CA) solutions. METHODS: The cytotoxicity of CA solutions in DMEM (diluted 1/10, 1/100) was assessed in L-929 fibroblasts. A broth macrodilution method (MIC and MBC) was used to assess CA antibacterial concentration. The antimicrobial activity of CA solutions was also evaluated after their final rinse inside root canals in previously Enterococcus faecalis-contaminated dentinal tubules. Ten infected dentine samples were rinsed for 5 min with 5% NaOCl and subsequently with 1% citric acid for 3 min. Another 10 were rinsed with 5% NaOCl and 10% citric acid for 3 min; the remaining four specimens were utilized as positive controls. Two uncontaminated specimens were used as negative controls. After LIVE/DEAD BacLight staining, the samples were assessed using CLSM to analyze the percentage of residual live and dead cells. RESULTS: Both undiluted and diluted CA solutions showed severe toxicity; no changes from normal morphology were displayed when diluted 1/100. The MIC and MBC of CA were 6.25 mg/mL and 12.50 mg/mL, respectively. CA solutions demonstrated significantly low levels of bacterial counts than the positive control group, reporting a value of 9.3% for the 10% solution versus the 1% solution (35.2%). CONCLUSIONS: Despite its valuable antimicrobial properties, the cytotoxic effects of citric acid should be considered during endodontic treatment.
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To investigate the dispersion and contamination of aerosols generated during coronal access performed by high-speed handpiece and ultrasonic device. To measure the aerosol dispersion, a red dye or an Enterococcus faecalis culture broth inside the bottle of the water system of the dental and ultrasonic unit were used. Bovine extracted teeth were allocated in six groups according to the coronal access: G1: diamond bur in high-speed handpiece (HS) with aspiration (A); G2: ultrasonic (US) inserts with aspiration; G3: combined coronal access with HS and US with aspiration; and G4, G5, and G6 were performed without aspiration (WA). The distance reached by the aerosol with the dye was measured in centimeters, and for environment contamination, agar-plates were arranged at standardized distances for counting colony-forming units (CFU/mL). The ANOVA followed by the Tukey tests were applied (α = 0.05). The coronal access with HS generated higher aerosol dispersion and contamination, even with simultaneous A (P < 0.05), while US generated less aerosol even WA (P < 0.05). The aspiration did not reduce the aerosol statistically. HS is a great source of aerosols in dental clinic during the coronal access and the use of US device should be encouraged.
Subject(s)
COVID-19 , Ultrasonics , Aerosols , Animals , Cattle , Humans , PandemicsABSTRACT
Seeking to increase the efficiency of endodontic irrigation, the association of different solutions as final irrigant has been investigated, such as sodium hypochlorite with chlorhexidine. The literature shows that the combination of these substances leads to the formation of a brownish precipitate, but does not reveal measurements of the intensity of this precipitate and its consequences. Aim: The present study aimed to evaluate the change in dentin color and the obliteration of the dentinal tubules after the association of sodium hypochlorite (NaOCl) with chlorhexidine (CHX) in the final irrigation. Methods: Fifty sterile human lower premolars were prepared with a ProDesign R 35.05 files and divided into 6 groups. Four different NaOCl concentrations (0.5%; 1%, 2.5% and 5.25%) associated with 2% CHX were tested, in addition to 2 control groups, using only 2.5% NaOCl and 2% CHX, respectively. After the final irrigation protocol, the dentin color change was evaluated by spectrophotometry immediately and after 24 hours, and the dentinal tubule obliteration was assessed by scanning electron microscopy. Results: It was possible to verify that regardless the NaOCl concentration used when associated with CHX, a chemical residue was formed, with consequent dentin pigmentation and tubular obstruction. There was a trend towards increased dentin pigmentation and tubular obstruction due to the deposition of the chemical residue formed by this association. Conclusion: It can be concluded that all concentrations of NaOCl associated with CHX caused color changes and tubular obstruction, being proportional to the concentration of NaOCl used
