Characterization of multidrug-resistant and virulent Klebsiella pneumoniae strains belonging to the high-risk clonal group 258 (CG258) isolated from inpatients in northeastern Brazil.

Multidrug-resistant (MDR) and hypervirulent Klebsiella pneumoniae (hvKp) clones have become a major threat to global public health. The clonal group 258 (CG258) is considered a high-risk CG and the K. pneumoniae strains belonging to it are often multi-resistant and to spread mainly in the hospital environment. This study aimed to characterize the antimicrobial resistance profile, virulence factors, and the clonal relationships among 13 K. pneumoniae strains belonging to CG258 from patients admitted to a tertiary hospital in Teresina, in the state of Piauí, northeastern Brazil. Ten strains were classified as MDR and three as extensively drug-resistant (XDR). Three different ß-lactamase-encoding genes (blaKPC, blaOXA-1-like, and blaCTX-M-Gp1) and six virulence genes (fimH, ycfM, mrkD, entB, ybtS, and kfu) were detected. Moreover, two hypermucoviscous K. pneumoniae strains and one capsular K-type 2 were found. Multilocus sequence typing analysis revealed ten different sequence types (STs) (ST14, ST17, ST20, ST29, ST45, ST101, ST268, ST1800, ST3995, and ST3996) belonging to CG258, being two (ST3995 and ST3996) described for the first time in this study.

Trans-chalcone and quercetin down-regulate fatty acid synthase gene expression and reduce ergosterol content in the human pathogenic dermatophyte Trichophyton rubrum.

BACKGROUND: Fatty acid synthase (FAS) is a promising antifungal target due to its marked structural differences between fungal and mammalian cells. The aim of this study was to evaluate the antifungal activity of flavonoids described in the scientific literature as FAS inhibitors (quercetin, trans-chalcone, ellagic acid, luteolin, galangin, and genistein) against the dermatophyte Trichophyton rubrum and their effects on fatty acid and ergosterol synthesis. METHODS: The antifungal activity of the natural products was tested by the microdilution assay for determination of the minimum inhibitory concentration (MIC). The effect of the compounds on the cell membrane was evaluated using a protoplast regeneration assay. Ergosterol content was quantified by spectrophotometry. Inhibition of FAS by flavonoids was evaluated by an enzymatic assay to determine IC50 values. Quantitative RT-PCR was used to measure transcription levels of the FAS1 and ERG6 genes involved in fatty acid and ergosterol biosynthesis, respectively, during exposure of T. rubrum to the flavonoids tested. RESULTS: The flavonoids quercetin and trans-chalcone were effective against T. rubrum, with MICs of 125 and 7.5 µg/mL for the wild-type strain (MYA3108) and of 63 and 1.9 µg/mL for the ABC transporter mutant strain (ΔTruMDR2), respectively. The MICs of the fluconazole and cerulenin controls were 63 and 125 µg/mL for the wild-type strain and 30 and 15 µg/mL for the mutant strain, respectively. Quercetin and trans-chalcone also reduced ergosterol content in the two strains, indicating that interference with fatty acid and ergosterol synthesis caused cell membrane disruption. The MIC of quercetin reduced the number of regenerated protoplasts by 30.26% (wild-type strain) and by 91.66% (mutant strain). Half the MIC (0.5 MIC) of quercetin did not reduce the number of regenerated wild-type fungal colonies, but caused a 36.19% reduction in the number of mutant strain protoplasts. In contrast, the MIC and 0.5 MIC of trans-chalcone and cerulenin drastically reduced protoplast regeneration in the two strains. The FAS1 gene was repressed in the presence of MICs of quercetin, trans-chalcone, fluconazole and cerulenin. The ERG6 gene was induced in the presence of MICs of fluconazole and cerulenin and was repressed in the presence of MICs of trans-chalcone and quercetin. Trans-chalcone and quercetin inhibited the enzymatic activity of FAS, with IC50 values of 68.23 and 17.1 µg/mL, respectively. CONCLUSION: Trans-chalcone and quercetin showed antifungal activity against T. rubrum, reducing ergosterol levels and modulating the expression of FAS1 and ERG6.

Genetic diversity among genotypes of Uncaria guianensis (Aubl.) J.F. Gmel. maintained in an in vitro germplasm bank.

Phytotherapeutic preparations from Uncaria guianensis (Aubl.) J.F. Gmel. (Rubiaceae) are marketed worldwide and are mainly used for their anti-inflammatory activity. The species has not yet been domesticated and is threatened by deforestation and overexploitation. It is, therefore, important to preserve and manage this genetic resource in germplasm banks, so that the extractive provision of plant material can be replaced by cultivated production. The aim of this study was to evaluate the genetic diversity among 20 genotypes maintained under in vitro conditions using 9 primers start codon targeted (SCoT) polymorphism, and to determine the concentrations of the pentacyclic oxindole alkaloids (POAs); mitraphylline and isomitraphylline in methanolic extracts by high-performance liquid chromatography (HPLC). Plantlets were cultivated on woody plant medium supplemented with 20 g.L-1 sucrose and 4.4 µM benzylaminopurine and incubated under a 16 h photoperiod for 45 days. SCoT analysis separated the genotypes into four divergent clusters and confirmed significant genetic diversity with up to 70% dissimilarity. Moreover, HPLC revealed considerable chemical variability and allowed the separation of the tested genotypes into high, medium and low producers of mitraphylline/isomitraphylline. Genotypes with the highest concentrations of POAs originated from the state of Acre and Amapá, while those with the lowest levels were from the state of Pará. The results demonstrate that the genetic diversity within the in vitro germplasm bank is sufficient to support breeding studies, selection of elite genotypes and the large-scale multiplication of plants that could serve as feedstock for the industrial-scale production of phytomedicines. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s13205-021-03016-y.

A novel hypermucoviscous Klebsiella pneumoniae ST3994-K2 clone belonging to Clonal Group 86.

Emergent hypervirulent Klebsiella pneumoniae has been responsible for severe diseases, representing a serious threat to public health. We report the whole-genome sequencing of a novel ST3994-K2 clone, a single locus variant of ST86 K2, which is considered a worrying hypervirulent clone that emerged in several parts of the world. The strain K. pneumonia Kpi144 was isolated in 2013 from a blood culture of a 69-year-old male patient admitted to a tertiary hospital in Teresina, state of Piauí, northeastern Brazil. The strain was susceptible to 41 antibiotics tested, presented hypermucoviscous phenotype and a virulent behavior was observed in the Galleria mellonella infection model. Moreover, the virulome showed several virulence genes. To the best of our knowledge, this is the first worldwide report of a novel ST3994-K2 K. pneumoniae clone, an SLV of ST86 K2, which is considered a worrying virulent clone that has emerged in several parts of the world, including South America and Brazil.

Green Synthesis, Characterization and Antimicrobial Evaluation of Silver Nanoparticles for an Intracanal Dressing.

Green chemistry has been applied in different areas due to the growing demands for renewable processes and one of them is nanotechnology. The aim of this study was to characterize a formulation containing silver nanoparticles (AgNPs) produced by a green synthesis and to evaluate its antimicrobial activity. The formulation will be used as an intracanal dressing exploiting the AgNPs' antimicrobial properties, which are crucial to prevent infections and bacterial reinfections that can compromise endodontic treatments. In the green synthesis, silver nitrate was employed as the precursor salt, maltose as a reducing agent, and gelatin as a stabilizing agent. The formulation was prepared mixing 50 % of a liquid containing the AgNPs and 50 % of hydroxyethylcellulose gel at 1.5 % with proper evaluation of the process inherent parameters. Techniques such as molecular absorption spectrometry and dynamic light scattering were used in characterization step. The antimicrobial activity of the AgNPs against Escherichia coli ATCC 25922, Enterococcus faecalis NCTC 775, Pseudomonas aeruginosa ATCC 27853, Staphylococcus aureus ATCC 25923 and Streptococcus mutans ATCC 25175 was verified according to National Comittee for Clinical Laboratory Standards (NCCLS) by determining minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC). The obtained results indicated the formulation containing AgNPs produced by a green synthesis was properly characterized by the selected techniques. Furthermore, the formulation assessment proved that it is suitable for the proposal as well as it has potential to be used as an intracanal dressing since presented antimicrobial activity against all bacterial strains evaluated.

Influence of solvents on the bond strength of resin sealer to intraradicular dentin after retreatment.

This study evaluated the removal of filling material with ProTaper Universal Rotary Retreatment system (PTR) combined with solvents and the influence of solvents on the bond strength (PBS) of sealer to intraradicular dentin after canal reobturation. Roots were endodontically treated and distributed to five groups (n = 12). The control group was not retreated. In the four experimental groups, canals were retreated with PTR alone or in combination with xylol, orange oil, and eucalyptol. After filling material removal, two specimens of each group were analysed by SEM and µCT to verify the presence of filling remnants on root canal walls. The other roots were reobturated and sectioned in 1-mm-thick dentin slices that were subjected to the push-out test. Data were analysed by two-way ANOVA and Tukey's test (α = 0.05). SEM and µCT analysis revealed that all retreatment techniques left filling remnants on canal walls. The control group (3.47 ± 1.21) presented significantly higher (p < 0.05) PBS than the experimental groups. The groups retreated with PTR alone (2.59 ± 0.99) or combined with xylol (2.54 ± 0.77) and orange oil (2.32 ± 0.93) presented similar bond strength (p > 0.05), and differed significantly from the group with eucalyptol (1.89 ± 0.63). The solvents reduced the PBS of the sealer to dentin and no retreatment technique promoted complete removal of filling material.

Effects of various irrigation/aspiration protocols on cleaning of flattened root canals.

This study aimed to evaluate, by means of scanning electron microscopy (SEM), the cleaning of flattened root canals, varying irrigation/aspiration protocols during biomechanical preparation. Thirdy human mandibular incisors were distributed into three groups (n = 10) according to the aspiration/irrigation protocols: conventional, conventional + brush, and apical negative pressure irrigation. Irrigation procedure was performed with 5 mL of 1% NaOCl at each change of instrument; final irrigation was conducted with 17% EDTA for 5 min. After biomechanical preparation, the roots were sectioned and prepared for SEM analysis. The images obtained were evaluated under 35× and 1,000× magnification by three calibrated examiners, following a double-blind design. All data were analyzed using Kruskal-Wallis and Dunn's post hoc tests (α = 0.05). Canals wherein the conventional method and apical negative pressure irrigation were employed revealed less debris, statistically different from the conventional + brush group (p < 0.05). Regarding the presence of smears, apical negative pressure irrigation was more effective in cleaning, showing lowest scores (p < 0.05), compared with the other tested protocols. Comparing each root canal third revealed that the apical portion was difficult to clean as all the tested protocols showed similar high scores (p > 0.05), both for the presence of debris and smear layer. In conclusion, although none of the studied irrigation/aspiration protocols have completely cleaned flattened root canals, apical negative pressure irrigation was more effective in smear layer removal, whereas the conventional + brush protocol was the least effective in removing the debris and smear layer.

Green synthesis, characterization and antimicrobial evaluation of silver nanoparticles for an intracanal dressing

Abstract Green chemistry has been applied in different areas due to the growing demands for renewable processes and one of them is nanotechnology. The aim of this study was to characterize a formulation containing silver nanoparticles (AgNPs) produced by a green synthesis and to evaluate its antimicrobial activity. The formulation will be used as an intracanal dressing exploiting the AgNPs' antimicrobial properties, which are crucial to prevent infections and bacterial reinfections that can compromise endodontic treatments. In the green synthesis, silver nitrate was employed as the precursor salt, maltose as a reducing agent, and gelatin as a stabilizing agent. The formulation was prepared mixing 50 % of a liquid containing the AgNPs and 50 % of hydroxyethylcellulose gel at 1.5 % with proper evaluation of the process inherent parameters. Techniques such as molecular absorption spectrometry and dynamic light scattering were used in characterization step. The antimicrobial activity of the AgNPs against Escherichia coli ATCC 25922, Enterococcus faecalis NCTC 775, Pseudomonas aeruginosa ATCC 27853, Staphylococcus aureus ATCC 25923 and Streptococcus mutans ATCC 25175 was verified according to National Comittee for Clinical Laboratory Standards (NCCLS) by determining minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC). The obtained results indicated the formulation containing AgNPs produced by a green synthesis was properly characterized by the selected techniques. Furthermore, the formulation assessment proved that it is suitable for the proposal as well as it has potential to be used as an intracanal dressing since presented antimicrobial activity against all bacterial strains evaluated.

Resumo A química verde tem sido aplicada em diferentes áreas devido à crescente demanda por processos renováveis e uma delas é a nanotecnologia. O objetivo deste estudo foi caracterizar uma formulação contendo nanopartículas de prata (AgNPs) produzidas por meio de síntese verde e avaliar sua atividade antimicrobiana. A formulação será usada como curativo intracanal explorando as propriedades antimicrobianas das AgNPs que são cruciais para prevenir infecções e reinfecções bacterianas que podem comprometer os tratamentos endodônticos. Na síntese verde, nitrato de prata foi empregado como sal precursor, maltose como agente redutor e gelatina como agente estabilizador. A formulação foi preparada misturando-se 50% do líquido contendo as AgNPs e 50% de gel de hidroxietilcelulose a 1,5% com avaliação adequada dos parâmetros inerentes ao processo. Técnicas como espectrometria de absorção molecular e espalhamento dinâmico de luz foram usadas na etapa de caracterização. A atividade antimicrobiana das AgNPs contra Escherichia coli ATCC 25922, Enterococcus faecalis NCTC 775, Pseudomonas aeruginosa ATCC 27853, Staphylococcus aureus ATCC 25923 e Streptococcus mutans ATCC 25175 foi verificada de acordo com o National Comittee for Clinical Laboratory Standards (NCCLS), determinando-se a concentração inibitória mínima (MIC) e a concentração bactericida mínima (MBC). Os resultados obtidos indicaram que a formulação contendo AgNPs produzidas por meio de síntese verde foi devidamente caracterizada pelas técnicas selecionadas. Além disso, a avaliação da formulação provou que ela é adequada para a proposta, bem como tem potencial para ser utilizada como curativo intracanal já que apresentou atividade antimicrobiana contra todas as cepas bacterianas avaliadas.

New methodology to evaluate bond strength of root-end filling materials.

This study evaluated the bond strength of root-end filling materials to root-end cavities using a new methodology. Twenty maxillary central incisors were subjected to biomechanical preparation (#80 hand file) and sectioned transversally 2 mm short of the apex and 4 mm coronally to this point. The root cylinders were embedded in acrylic resin and positioned at 45° to the horizontal plane for preparation of root-end cavities with a diamond ultrasonic retrotip. Two groups (n=10) were formed according to the root-end filling material: MTA and Super EBA. A gutta-percha cone (#80) was tug-backed at the limit between the canal and the root-end cavity. The cavity was filled and the gutta-percha cone was removed after complete setting of the sealer. The specimens were placed in an Instron machine with the root-end filling turned downwards. The push-out shaft was inserted in the space previously occupied by the gutta-percha cone and run at a crosshead speed of 1.0 mm/min for pushing out the root-end filling material. Data were analyzed by ANOVA (α=5%). Super EBA (6.03±1.31) presented higher bond strength (MPa) than MTA (1.81±0.45) (p>0.05). There was a predominance of cohesive failures for Super EBA and mixed for MTA. The protocol of specimen preparation is effective and introduces a specific methodology for assessing bond strength of root-end filling materials to dentin. Among the materials, Super EBA presented the highest bond strength.

Protection against T1DM-Induced Bone Loss by Zinc Supplementation: Biomechanical, Histomorphometric, and Molecular Analyses in STZ-Induced Diabetic Rats.

Several studies have established an association between diabetes and alterations in bone metabolism; however, the underlying mechanism is not well established. Although zinc is recognized as a potential preventive agent against diabetes-induced bone loss, there is no evidence demonstrating its effect in chronic diabetic conditions. This study evaluated the effects of zinc supplementation in a chronic (90 days) type 1 diabetes-induced bone-loss model. Male Wistar rats were distributed in three groups: control, type 1 diabetes mellitus (T1DM), and T1DM plus zinc supplementation (T1DMS). Serum biochemical analysis; tibia histomorphometric, biomechanical, and collagen-content analyses; and femur mRNA expression were evaluated. Relative to T1DM, the zinc-supplemented group showed increased histomorphometric parameters such as TbWi and BAr and decreased TbSp, increased biomechanical parameters (maximum load, stiffness, ultimate strain, and Young's modulus), and increased type I collagen content. Interestingly, similar values for these parameters were observed between the T1DMS and control groups. These results demonstrate the protective effect of zinc on the maintenance of bone strength and flexibility. In addition, downregulation of OPG, COL1A, and MMP-9 genes was observed in T1DMS, and the anabolic effects of zinc were evidenced by increased OC expression and serum ALP activity, both related to osteoblastogenesis, demonstrating a positive effect on bone formation. In contrast, T1DM showed excessive bone loss, observed through reduced histomorphometric and biomechanical parameters, characterizing diabetes-associated bone loss. The bone loss was also observed through upregulation of OPG, COL1A, and MMP-9 genes. In conclusion, zinc showed a positive effect on the maintenance of bone architecture and biomechanical parameters. Indeed, OC upregulation and control of expression of OPG, COL1A, and MMP-9 mRNAs, even in chronic hyperglycemia, support an anabolic and protective effect of zinc under chronic diabetic conditions. Furthermore, these results indicate that zinc supplementation could act as a complementary therapy in chronic T1DM.

Bidens pilosa - picão preto: influência da adubação orgânica e da luminosidade na produtividade e no teor de flavonoides / Bidens pilosa - hairy beggartick: influence of organic fertilization and luminosity on productivity and flavonoid content

Espécies vegetais podem ser consideradas úteis, ou não, dependendo do modo que são caracterizadas pelos humanos. Algumas passam de daninhas a alimentícias, cosméticas ou medicinais, tornando interessante economicamente seu cultivo. Bidens pilosa L. (picão preto) é uma espécie, neste contexto, que foi submetida a ensaio agronômico visando avaliar sua produtividade (massa fresca e massa seca) e o teor de flavonoides. Ela foi cultivada com diferentes níveis de adubação orgânica (esterco bovino curtido - 50 t ha-1) e de sombreamento. O delineamento experimental foi de blocos casualizados com 3 repetições, em esquema fatorial de 22, constituído de 4 tratamentos (1. Com adubação + sombreamento 0%; 2. Com adubação + sombreamento 50%; 3. Sem adubação + sombreamento 50% e 4. Sem adubação + sombreamento 0%). Os resultados mostraram que o cultivo usando adubação orgânica, ao sol pleno produziu 63% mais droga vegetal que os tratamentos sem adubação orgânica, usando ou não sombreamento. Em relação ao teor de flavonoides, igualdade estatística entre todos os tratamentos foi encontrada, porém o maior resultado numérico (0,096% m/m) para este parâmetro foi obtido sem uso de adubação e sem restrição de iluminação e o menor (0,058% m/m), com uso de adubação ao sol pleno.

Isolation and characterization of a Pseudomonas aeruginosa from a virgin Brazilian Amazon region with potential to degrade atrazine.

The use of pesticides to increase agricultural production can result in the contamination of the environment, causing changes in the genetic structure of organisms and in the loss of biodiversity. This practice is also inducing changes in the rainforest ecosystem. In this work, a Pseudomonas aeruginosa isolated from a preservation soil area of the Brazilian Amazon Forest, without usage of any pesticide, was evaluated for its potential to degrade atrazine. This isolate presented all responsible genes (atzA, atzB, atzC, atzD, atzE, and atzF) for atrazine mineralization and demonstrated capacity to use atrazine as a nitrogen source, having achieved a reduction of 44 % of the initial concentration of atrazine after 24 h. These results confirm gene dispersion and/or a possible contamination of the area with the herbicide, which reinforces global concern of the increase and intensive use of pesticides worldwide.

Development of intracanal formulation containing silver nanoparticles.

This study aimed to synthetize, characterize and evaluate the antimicrobial properties of silver nanoparticles to be used in the development of a root intracanal formulation. Silver nanoparticles (AgNPs) were obtained by reduction of silver nitrate with sodium borohydride and characterized by UV-Visible spectrophotometry, scanning electron microscopy (SEM) and dynamic light scattering (DLS). The antimicrobial activity of nanoparticle formulation was evaluated by determinations of the minimum inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) against different bacterial species by the microdilution method, according to recommendations of the Clinical and Laboratory Standards Institute (CLSI). Three potential vehicles, hydroxyethylcellulose, Carbomer and polyethylene glycol were tested as carriers for formulations containing AgNPs. The efficiency of the synthesis method chosen to produce AgNPs was demonstrated by four characterization techniques. The nanoparticles showed antibacterial activity against all species tested. Incorporation of AgNPs into all experimental vehicles produced stable formulations but the one in hydroxyethylcellulose presented better physical proprieties. The results indicate that silver nanoparticles are potential antiseptic agents to be used in root canals and incorporation in adequate vehicles may favor a broader application.

Quantitative assessment of root canal roughness with calcium-based hypochlorite irrigants by 3D CLSM.

Chemical solutions play important roles in endodontic treatment and promote ultrastructural changes in dentin surface. The aim of this study was to quantify root canal roughness at different concentrations of calcium hypochlorite (Ca(OCl)2) and sodium hypochlorite (NaOCl) by confocal laser scanning microscopy (CLSM). Fifty-two human mandibular premolars were sectioned and randomly organized into thirteen groups (n=8): saline (control); 1%, 2.5% and 5% NaOCl; 1%, 2.5% and 5% Ca(OCl)2; the hypochlorite groups were further divided into with or without EDTA. The chlorine concentrations of the different solutions were measured by iodine titration (%). The superficial roughness (Sa) was quantified by CLSM. Ca(OCl)2 presented substantial decrease in chlorine concentration that differed from the package indication, but without compromising the dentin ultrastructure changes. There were no significant differences in dentin roughness between Ca(OCl)2 or NaOCl at all studied concentrations. The combination with EDTA provided similar roughness values among the solutions (p>0.05). The 5% Ca(OCl)2 and NaOCl solutions significantly increased dentin roughness and did not differ from the EDTA association (p>0.05). Ca(OCl)2 promoted similar dentin roughness as the NaOCl at the same concentrations and combined with EDTA. It may be concluded that Ca(OCl)2 modified the root canal dentin roughness similarly to NaOCl, at the same concentrations and EDTA combinations used in this study. Ca(OCl)2 and NaOCl, both at 5%, significantly altered dentin roughness, overcoming EDTA association, thus Ca(OCl)2 concentrations ranging from 1% to 2.5% may be suitable solutions for root canal irrigation protocols.

Physicochemical properties of endodontic sealers of different bases.

OBJECTIVE: To assess the setting time (ST), flow (FL), radiopacity (RD), solubility (SB) and dimensional change following setting (DC) of different sealers (AH Plus®, Polifil, Apexit Plus®, Sealapex®, Endométhasone® and Endofill®) according to American National Standards Institute/American Dental Association (ANSI/ADA) Specification 57. MATERIAL AND METHODS: Five samples of each material were used for each test. For ST, cast rings were filled with sealers and tested with a Gilmore needle. For FL, the sealer was placed on a glass plate. After 180 s, another plate with 20 g and a load of 100 g were applied on the material, and the diameters of the discs formed were measured. In RD, circular molds were filled with the sealers, radiographed and analyzed using Digora software. For SB, circular molds were filled with the sealers, a nylon thread was placed inside the material and another glass plate was positioned on the set, pressed and stored at 37°C. Samples were weighed, placed in water, dried and reweighed. The water used for SB was analyzed by atomic absorption spectrometry. For DC, circular molds were filled with the sealers, covered by glass plates and stored at 37°C. Samples were measured and stored in water for 30 days. After this period, they were dryed and measured again. RESULTS: Regarding ST, AH Plus®, Apexit® and Endofil® sealers are in accordance with ANSI/ADA standards. Endométhasone's manufacturer did not mention the ST; Polifil is an experimental sealer and Sealapex® did not set. Considering RD, SB and DC, all sealers were in accordance with ANSI/ADA. The spectrometric analysis showed that a significant amount of K+ and Zn2+ ions was released from Apexit Plus® and Endofill®, respectively. CONCLUSION: Except for DC, all other physicochemical properties of the tested sealers conformed to ANSI/ADA requirements.

Influence of solvents on the bond strength of resin sealer to intraradicular dentin after retreatment

Abstract This study evaluated the removal of filling material with ProTaper Universal Rotary Retreatment system (PTR) combined with solvents and the influence of solvents on the bond strength (PBS) of sealer to intraradicular dentin after canal reobturation. Roots were endodontically treated and distributed to five groups (n = 12). The control group was not retreated. In the four experimental groups, canals were retreated with PTR alone or in combination with xylol, orange oil, and eucalyptol. After filling material removal, two specimens of each group were analysed by SEM and µCT to verify the presence of filling remnants on root canal walls. The other roots were reobturated and sectioned in 1-mm-thick dentin slices that were subjected to the push-out test. Data were analysed by two-way ANOVA and Tukey’s test (α = 0.05). SEM and µCT analysis revealed that all retreatment techniques left filling remnants on canal walls. The control group (3.47 ± 1.21) presented significantly higher (p < 0.05) PBS than the experimental groups. The groups retreated with PTR alone (2.59 ± 0.99) or combined with xylol (2.54 ± 0.77) and orange oil (2.32 ± 0.93) presented similar bond strength (p > 0.05), and differed significantly from the group with eucalyptol (1.89 ± 0.63). The solvents reduced the PBS of the sealer to dentin and no retreatment technique promoted complete removal of filling material.

Radiopacity evaluation of Portland and MTA-based cements by digital radiographic system.

OBJECTIVE: The aim of the present study was to evaluate the radiopacity of Portland and MTA-based cements using the Digora TM digital radiographic system. MATERIAL AND METHODS: The performed tests followed specification number 57 from the American National Standard Institute/American Dental Association (2000) for endodontic sealing materials. The materials were placed in 5 acrylic plates, especially designed for this experiment, along with a graduated aluminum stepwedge varying from 1 to 10 mm in thickness. The set was radiographed at a 30 cm focus-object distance and with 0.2 s exposure time. After the radiographs were taken, the optical laser readings of radiographs were performed by Digora TM system. Five radiographic density readings were performed for each studied material and for each step of the aluminum scale. RESULTS: White ProRoot MTA (155.99±8.04), gray ProRoot MTA (155.96±16.30) and MTA BIO (143.13±16.94) presented higher radiopacity values (p<0.05), while white non-structural Portland (119.76±22.34), gray Portland (109.71±4.90) and white structural Portland (99.59±12.88) presented lower radiopacity values (p<0.05). CONCLUSIONS: It was concluded that MTA-based cements were the only materials presenting radiopacity within the ANSI/ADA specifications.

Influence of apical enlargement in cleaning and extrusion in canals with mild and moderate curvatures.

Optical microscopy and morphometric analysis were used in this study to evaluate, in vitro, the cleaning of the apical region in root canals with mild or moderate curvatures subjected to biomechanical preparation with a rotary system, as well as to assess the amount of extruded material to the periapical area. Lateral incisors (n = 32), 16 with curvature angles smaller or equal to 10º (GI) and 16 between 11º and 25º angles (GII) were submitted to Hero 642 rotary instrumentation with different surgical diameters: (A) 30.02 and (B) 45.02. Irrigation was performed at each change of instrument with 5 mL of ultrapure Milli-Q water and the extruded material through the apical foramen was collected. Root cross-sections were subjected to histological analysis by optical microscopy (×40) and the images were evaluated morphometrically using the Image Tool software. Quantification of the extruded material was performed by weighing after liquid evaporation. ANOVA showed no statistically significant differences (p>0.05) among the groups with respect to the procedures used to clean the apical region. Considering the amount of extruded material, the Tukey's HSD showed that canals with mild curvature prepared with the 45.02 surgical diameter showed significantly higher values (p<0.05) that those of the other groups, which were similar between themselves (p>0.05). In conclusion, the effect of cleaning the apical region did not differ in the groups, considering root curvature and the surgical diameter of instruments used for apical preparation. The amount of extruded material was greater in canals with mild curvature that were prepared with the 45.02 surgical instrument diameter.

Evaluation of physico-chemical properties of Portland cements and MTA.

The purpose of this study was to evaluate the hydrogenionic potential and electrical conductivity of Portland cements and MTA, as well as the amount of arsenic and calcium released from these materials. In Teflon molds, samples of each material were agitated and added to plastic flasks containing distilled water for 3, 24, 72 and 168 h. The results were analyzed with a Kruskal-Wallis non-parametric test for global comparisons and a Dunn-Tukey test for pairwise comparisons. The results revealed no significant differences in the pH of the materials (p > 0.05). The electrical conductivity of the cements were not statistically different (p > 0.05). White non-structural cement and MTA BIO released the largest amount of calcium ions into solution (p < 0.05), while arsenic release was insignificant in all of the materials (p > 0.05). The results indicated that the physico-chemical properties of Portland cements and MTA were similar. Furthermore, all materials produced an alkaline environment and can be considered safe for clinical use because arsenic was not released. The electrical conductivity and the amount of calcium ions released into solution increased over time.

Physicochemical properties of methacrylate resin-based root canal sealers.

INTRODUCTION: This study assessed in vitro the physicochemical properties of 2 methacrylate resin-based sealers (Epiphany SE and Hybrid Root SEAL), comparing the results with a well-established epoxy resin-based sealer (AH Plus). METHODS: Five samples of each material were used for each test (setting time, flow, radiopacity, dimensional change after setting, and solubility) according to American National Standards Institute/American Dental Association (ANSI/ADA) Specification 57. The samples were assigned to 3 groups: I, AH Plus; II, Epiphany SE; and III, Hybrid Root SEAL. The distilled and deionized water used at the solubility test was submitted to atomic absorption spectrometry to observe the presence of Ca2+, K+, Ni2+, and Zn2+ ions. In addition, the surface morphology of the specimens was analyzed by means of scanning electron microscopy (SEM). Statistical analysis was performed by using one-way analysis of variance and Tukey-Kramer test (P < .05). RESULTS: Flow, radiopacity, and solubility of all sealers were in accordance with ANSI/ADA. The setting time of Hybrid Root SEAL did not agree with ANSI/ADA requirements. The dimensional change of all sealers was greater than the values considered acceptable by ANSI/ADA. The spectrometry analysis showed significant Ca(2+) ions release for AH Plus. In SEM analysis, Hybrid Root SEAL presented spherical monomers with inferior size than AH Plus and Epiphany SE. CONCLUSIONS: It might be concluded that physicochemical properties of the tested sealers conformed to ANSI/ADA (2000) standardization, except for the setting time of Hybrid Root SEAL and the dimensional change of all sealers, which did not fulfill the ANSI/ADA requirements.