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1.
Rev. Fac. Odontol. (B.Aires) ; 36(84): 21-26, 2021. tab
Article in Spanish | LILACS | ID: biblio-1363852

ABSTRACT

La terapia endodóntica tiene como uno de sus objetivos lograr la completa desinfección del sistema de conductos radiculares. Por esto, se deben seleccionar sustancias irrigantes que tengan la capacidad de eliminar todo el contenido de dicho sistema. La acción antimicrobiana es una de las características más importantes a tener en cuenta en la elección. El hipoclorito de sodio (NaOCl) tiene capacidad bactericida sobre muchos de los microorganismos de la flora endodóntica. El Enterococcus faecalis es una bacteria altamente resistente a antibacterianos que sobrevive en condiciones extremas. El ácido hipocloroso (HOCl) es una molécula derivada del NaOCl que ha demostrado tener alto poder bactericida sobre cepas patogénicas bucales. El objetivo de este trabajo fue evaluar y comparar la efectividad antimicrobiana in vitro del NaOCl 2.5% y el HOCl al 5% frente a Enterococcus faecalis. Una suspensión de Enterococcus faecalis (ATCC29212), de turbidez 0.5 en escala de McFarland, fue inoculada en varios tubos de ensayo, los cuales contenían cada antimicrobiano. Se dejaron actuar durante 1, 5 y 10 minutos para luego neutralizarlos e inclubarlos a 37º C en condiciones de capnofilia durante 48 hs. Todo el procedimiento se realizó por quintuplicado. Los resultados se midieron mediante recuento de UFC/ml. No se evidenció presencia de Enterococcus faecalis en las placas que contenían la solución de NaOCl al 2.5% como tampoco en aquellas que contenían HOCl al 5%. In vitro, el HOCl y el NaOCl en las concentraciones probadas, eliminaron completamente las cepas de Enterococcus faecalis (AU)


Subject(s)
Sodium Hypochlorite/therapeutic use , Enterococcus faecalis/drug effects , Hypochlorous Acid/therapeutic use , Anti-Bacterial Agents/therapeutic use , Root Canal Irrigants/therapeutic use , In Vitro Techniques , Colony Count, Microbial , Culture Media , Dental Pulp Cavity/microbiology
2.
Rev. Asoc. Odontol. Argent ; 108(2): 46-51, mayo-ago. 2020. tab
Article in Spanish | LILACS | ID: biblio-1121108

ABSTRACT

Objetivos: Comparar ex vivo la eficacia del instrumento XP-endo Finisher y del sistema EndoActivator en la reducción/eliminación del biofilm microbiano en conductos radiculares infectados. Materiales y métodos: Se utilizaron 23 premolares inferiores humanos extraídos cuya longitud fue estandarizada en 17 mm. Todos los conductos se prepararon con el sistema WaveOne Gold Medium (#35.06). Los dientes se esterilizaron, se inocularon con Enterococcus faecalis y se separaron en dos grupos experimentales de 10 piezas cada uno. De los 3 dientes remanentes, 1 fue utilizado como control positivo y 2, como controles negativos. En el grupo 1, las soluciones irrigantes se agitaron con XP-endo Finisher. En el grupo 2, se utilizó EndoActivator. Se tomaron muestras antes de la contaminación, luego de esta y después de la agitación de los irrigantes mediante conos de papel estériles. La carga microbiana fue sembrada en agar sangre y los conos se cultivaron en caldo tripteína de soja. La remoción de la carga microbiana se determinó por la presencia o ausencia de turbiedad del medio. Las unidades formadoras de colonias (UFC) remanentes se cuantificaron y los resultados se categorizaron como R1 (≤10 UFC) o R2 (>10 UFC). Los datos fueron analizados mediante la prueba de Fisher. Resultados: No hubo diferencias significativas entre XP-endo Finisher y EndoActivator (P>0,05). El número de usos no influyó sobre la capacidad operativa de ambos instrumentos (AU)


Aim: To compare ex vivo the effectiveness of the XP-endo Finisher and the EndoActivator in biofilm reduction/ removal from infected root canals. Materials and methods: Twenty three extracted human single-rooted lower premolars were selected and standardised to 17 mm in length. All the canals were prepared with WaveOne Gold Medium reciprocating files (#35.06). The teeth were autoclaved and inoculated with Enterococcus faecalis. The infected teeth were then assigned to 2 experimental groups of 10 teeth each according to the final irrigation/agitation protocol. Of the three remaining teeth, one was used as a positive control, and the other two were used as negative controls. In Group 1 the irrigating solutions were agitated with XP-endo Finisher while in Group 2 the EndoActivator was used. All root canals were sampled before and after contamination, and again after irrigant agitation with sterile paper points. The microbial load was spread on blood agar plates and the paper points were cultured in sterile trypticase soy broth. The removal of the microbial load was determined by visual observation of the turbidity of the media and by quantification of the number of colony-forming units (UFC). The results were categorized as R1 (≤10 UFC) or R2 (>10 UFC). Data were analysed by the Fisher's exact test at P<0.05. Results: No significant differences was found between XP-endo Finisher and EndoActivator (P>0.05) regarding their effectiveness in the reduction/removal of the microbial biofilm. The number of uses of both instruments did not affect their operative performance (AU) Conclusion: XPF and EA were both equally effective for microbial biofilm reduction/removal from ex vivo infected root canals (AU)


Subject(s)
Root Canal Irrigants/chemistry , Dental High-Speed Equipment , Biofilms , Dental Instruments , Dental Pulp Cavity/microbiology , In Vitro Techniques , Colony Count, Microbial/methods , Efficacy , Statistical Analysis , Enterococcus faecalis/isolation & purification , Culture Media
3.
Rev. Cient. CRO-RJ (Online) ; 5(2): 69-77, May-Aug. 2020.
Article in English | LILACS, BBO | ID: biblio-1254137

ABSTRACT

Introduction: Description of the bacterial community before and after chemomechanical preparation (CP) with the removal of a smear layer (SL) in pulpectomized primary teeth has been little reported. Objective: These case reports describe the presence of total microorganisms and Enterococcus faecalis in root canals of primary incisors before and after CP with SL removal. Case Reports: Microbiological samples were collected from the root canals of three children (3.66±0.58 years old) with necrosis (n=2) and irreversible pulpal inflammation (n=1) in maxillary primary incisors. After teeth isolation with a rubber dam and antisepsis of the operative field, the sample collections were performed with sterile absorbent paper cones before and after the CP, which included irrigation with 2.5% sodium hypochlorite followed by 6% citric acid to remove the SL. The collected samples were analysed immediately at the end of the clinical procedures. The plates were incubated anaerobically for 48 hours at 37°C. The results were expressed as colony forming units (CFU)/mL. Results: Two of the three teeth showed total microorganisms before the CP. One incisor had no microorganisms in the initial collection. No CFU was counted in the samples collected after CP. Moreover, E. faecalis was not observed any time, either before or after the CP. Conclusions: E. faecalis was not detected in any sample, yet two of the three root canals had microorganisms before CP. In cases where microorganisms were initially found, 100% elimination was observed after the applied protocol.


Introdução: A descrição da comunidade bacteriana antes e após o preparo químico-mecânico (PQM) com remoção da smear layer (SL) em dentes decíduos pulpectomizados tem sido pouco relatada. Objetivo: Esses relatos de casos descrevem a presença de microrganismos totais e Enterococcus faecalis em canais radiculares de incisivos decíduos antes e após PQM com remoção de SL. Relatos dos Casos: Amostras microbiológicas foram coletadas do canal radicular de três crianças (3,66 ± 0,58 anos) com necrose (n = 2) e inflamação pulpar irreversível (n= 1) em incisivos decíduos superiores. Após o isolamento dos dentes com dique de borracha e antissepsia do campo operatório, as coletas das amostras foram realizadas com cones de papel absorvente estéril antes e após o PQM, que incluiu irrigação com hipoclorito de sódio 2,5% seguido de ácido cítrico 6% para retirada do SL. As amostras coletadas foram analisadas imediatamente ao final dos procedimentos clínicos. As placas foram incubadas em anaerobiose durante 48 horas a 37°C. Os resultados foram expressos em unidades formadoras de colônias (UFC)/mL. Resultados: Dois dos três dentes apresentaram microrganismos totais antes do PQM. Um incisivo não apresentava microrganismos na coleta inicial. Nenhuma UFC foi contada nas amostras coletadas após o PQM. Além disso, o E. faecalis não foi observado nenhum momento, nem antes, nem depois do PQM. Conclusão: Não foi detectado E. faecalis em nenhuma amostra, porém dois dos três canais radiculares apresentavam microrganismos antes do PQM. Nos casos em que foram encontrados microrganismos inicialmente, observou-se 100% de eliminação após o protocolo aplicado.


Subject(s)
Humans , Male , Female , Child, Preschool , Pulpectomy/methods , Smear Layer/microbiology , Dental Pulp Necrosis/microbiology , Dental Pulp Cavity/microbiology
4.
Int. j. odontostomatol. (Print) ; 14(3): 448-456, 2020. tab, graf
Article in English | LILACS | ID: biblio-1114920

ABSTRACT

Enterococci are important nosocomial pathogens due to their intrinsic multiresistance and the acquisition of new antibiotic resistance genes (ARG). Enterococcus faecalis has been shown to be one of the main pathogens in persistent endodontic infections, therefore, the main objective of this study was to evaluate the phenotype and resistance genotype of strains of E. faecalis isolated from teeth with persistent endodontic lesions, to the most commonly prescribed antibiotics in dentistry. Thirteen strains of E. faecalis of different pulsotype were analyzed to evaluate the susceptibility to antibiotics, amoxicillin, amoxicillin/clavulanic acid, tetracycline, erythromycin and metronidazole, using the Epsilometer test (E- test) and the presence of beta-lactamases with nitrocefin test. Finally, the detection of ARG was performed with a molecular polymerase chain reaction (PCR) technique and confirmed by the sequencing of the amplification products. Fisher's exact test was used, using 95 % confidence. Regarding the phenotype of resistance, the evaluated strains, independent of the pulsotype, were totally resistant to the action of metronidazole. Antibiotics with higher minimum inhibitory concentration (MIC) after metronidazole include tetracycline and erythromycin. In contrast, lower MIC are applied to the combination of amoxicillin with clavulanic acid. The nitrocefin test was positive only in one strain. Genotypically, two genetically distant strains isolated from a single patient, presented a genotype of resistance to erythromycin, determined by the presence of the ermB gene. No statistically significant relationship was found between phenotypic resistance and the presence of ARG in relation to erythromycin (p> 0.05). It was concluded that isolates of E. faecalis from persistent endodontic infections showed phenotypes of resistance to several antimicrobial agents, all of which were susceptible to amoxicillin/clavulanic acid. Periodic evaluation of susceptibility to antibiotics is suggested as an important practice for the surveillance of antibiotic resistance in oral strains.


Los enterococos son importantes patógenos nosocomiales debido a su multi resistencia intrínseca y la adquisición de nuevos genes de resistencia a los antibióticos (ARG). Enterococcus faecalis es uno de los principales patógenos en infecciones endodónticas persistentes, por lo tanto, el objetivo principal de este estudio fue evaluar el fenotipo y el genotipo de resistencia de cepas de E. faecalis aisladas de dientes con lesiones endodóncicas persistentes, a los antibióticos comúnmente recetados en odontología. Se analizaron 13 cepas de E. faecalis de diferentes pulsotipos para evaluar la susceptibilidad a los antibióticos, amoxicilina, amoxicilina / ácido clavulánico, tetraciclina, eritromicina y metronidazol, utilizando la prueba de Epsilometría (E-test) y la presencia de beta-lactamasas con prueba de nitrocefina. Finalmente, la detección de ARG se realizó con una técnica molecular de reacción en cadena de la polimerasa (PCR) y se confirmó mediante la secuenciación de los productos de amplificación. Se utilizó la prueba exacta de Fisher, con un 95 % de confianza. En cuanto al fenotipo de resistencia, las cepas evaluadas, independientes del pulsotipo, fueron totalmente resistentes a la acción del metronidazol. Los antibióticos con los valores más altos de concentración mínima inibitoria (CMI) después del metronidazol incluyen tetraciclina y eritromicina. En contraste, las CMI mas bajas se aplican a la combinación de amoxicilina con ácido clavulánico. La prueba de nitrocefina fue positiva solo en una cepa. Genotípicamente, dos cepas distantes genéticamente, aisladas de un mismo paciente fueron positivas para el gen ermB. No se encontró una relación estadísticamente significativa entre la resistencia fenotípica y la presencia de ARG en relación con la eritromicina (p> 0,05). Se concluyó que los aislamientos de E. faecalis de infecciones endodónticas persistentes mostraron fenotipos de resistencia a varios agentes antimicrobianos, todos los cuales fueron susceptibles a amoxicilina / ácido clavulánico. Se sugiere una evaluación periódica de la susceptibilidad a los antibióticos como una práctica importante para la vigilancia de la resistencia a los antibióticos en las cepas orales.


Subject(s)
Humans , Enterococcus faecalis/drug effects , Enterococcus faecalis/genetics , Dental Pulp Cavity/microbiology , Anti-Bacterial Agents/pharmacology , Tetracycline , Microbial Sensitivity Tests , Erythromycin , Polymerase Chain Reaction , Clavulanic Acid/pharmacology , Drug Resistance, Bacterial/genetics , Amoxicillin/pharmacology , Metronidazole
5.
J. appl. oral sci ; 28: e20190100, 2020. tab, graf
Article in English | LILACS, BBO | ID: biblio-1056594

ABSTRACT

Abstract Objective: This clinical study sought to evaluate the effectiveness of passive ultrasonic activation (PUA) in eliminating microorganisms in primary endodontic infection (PEI) after instrumentation of root canals using microbiological culture and checkerboard DNA-DNA hybridization. Methodology: Twenty root canals with PEI and apical periodontitis were selected. The root canals were instrumented and then randomly divided into 2 groups, according to the irrigation method: PUA and conventional needle irrigation (CNI). Microbiological samples were collected before instrumentation (S1), after instrumentation (S2) and after irrigation with 17% EDTA (S3). The samples were subjected to anaerobic culture technique and checkerboard DNA-DNA hybridization analysis. Results: A statistically significant difference was found between CNI (23.56%) and PUA (98.37%) regarding the median percentage values for culturable bacteria reduction (p<0.05). In the initial samples, the most frequently detected species was S. constellatus (50%), and after root canal treatment was E. faecalis (50%). Conclusion: Both treatments significantly decreased the number of bacterial species compared with the initial sample. However, no statistical difference in the total microbial load between PUA and CNI groups was detected. The number of cultivable anaerobic bacteria reduced significantly using PUA, and the bacterial composition and number of bacterial species after using either CNI or PUA was similar.


Subject(s)
Humans , Male , Female , Adolescent , Adult , Middle Aged , Periapical Periodontitis/therapy , Root Canal Therapy/instrumentation , Ultrasonic Therapy/instrumentation , Dental Pulp Cavity/microbiology , Root Canal Irrigants/therapeutic use , Root Canal Therapy/methods , Sodium Hypochlorite/therapeutic use , Bacteria/isolation & purification , Ultrasonic Therapy/methods , Colony Count, Microbial , DNA Probes , Linear Models , Analysis of Variance , Treatment Outcome , Cone-Beam Computed Tomography , Bacterial Load , Therapeutic Irrigation/instrumentation , Therapeutic Irrigation/methods
6.
J. appl. oral sci ; 28: e20190699, 2020. graf
Article in English | LILACS, BBO | ID: biblio-1134770

ABSTRACT

Abstract Purpose To evaluate the kinetics of apical periodontitis development in vivo , induced either by contamination of the root canals by microorganisms from the oral cavity or by inoculation of bacterial lipopolysaccharide (LPS) and the regulation of major enzymes and receptors involved in the arachidonic acid metabolism. Methodology Apical periodontitis was induced in C57BL6 mice (n=96), by root canal exposure to oral cavity (n=48 teeth) or inoculation of LPS (10 µL of a suspension of 0.1 µg/µL) from E. coli into the root canals (n= 48 teeth). Healthy teeth were used as control (n=48 teeth). After 7, 14, 21 and 28 days the animals were euthanized and tissues removed for histopathological and qRT-PCR analyses. Histological analysis data were analyzed using two-way ANOVA followed by Sidak's test, and qRT-PCR data using two-way ANOVA followed by Tukey's test (α=0.05). Results Contamination by microorganisms led to the development of apical periodontitis, characterized by the recruitment of inflammatory cells and bone tissue resorption, whereas inoculation of LPS induced inflammatory cells recruitment without bone resorption. Both stimuli induced mRNA expression for cyclooxygenase-2 and 5-lipoxygenase enzymes. Expression of prostaglandin E 2 and leukotriene B 4 cell surface receptors were more stimulated by LPS. Regarding nuclear peroxisome proliferator-activated receptors (PPAR), oral contamination induced the synthesis of mRNA for PPARδ, differently from inoculation of LPS, that induced PPARα and PPARγ expression. Conclusions Contamination of the root canals by microorganisms from oral cavity induced the development of apical periodontitis differently than by inoculation with LPS, characterized by less bone loss than the first model. Regardless of the model used, it was found a local increase in the synthesis of mRNA for the enzymes 5-lipoxygenase and cyclooxygenase-2 of the arachidonic acid metabolism, as well as in the surface and nuclear receptors for the lipid mediators prostaglandin E2 and leukotriene B4.


Subject(s)
Animals , Male , Periapical Periodontitis/microbiology , Dinoprostone/metabolism , Lipopolysaccharides/metabolism , Leukotriene B4/metabolism , Dental Pulp Cavity/microbiology , Periapical Periodontitis/metabolism , Periapical Periodontitis/pathology , Time Factors , Bone Resorption/metabolism , Bone Resorption/microbiology , Arachidonate 5-Lipoxygenase/analysis , Arachidonate 5-Lipoxygenase/metabolism , RNA, Messenger/analysis , RNA, Messenger/metabolism , Dinoprostone/analysis , Random Allocation , Gene Expression , Leukotriene B4/analysis , Reverse Transcriptase Polymerase Chain Reaction , Dental Pulp Cavity/metabolism , Dental Pulp Cavity/pathology , Cyclooxygenase 2/analysis , Cyclooxygenase 2/metabolism , Mice, Inbred C57BL
7.
J. appl. oral sci ; 27: e20180256, 2019. tab
Article in English | LILACS, BBO | ID: biblio-1012514

ABSTRACT

Abstract Objective The rDNA-based method is unable to distinguish between alive and dead cells. Alternatively, bacterial viability can be assessed by molecular methods based on ribosomal RNA (rRNA). Therefore, this study aimed to detect viable streptococci in root canal samples using rRNA-based reverse transcription polymerase chain reaction (RT-PCR), compared to an rDNA-based PCR assay. Methodology Microbiological root canal samples were obtained from 32 teeth with primary endodontic infections before (S1) and after chemomechanical preparation (S2), and after removal of intracanal medication (S3). RNA and DNA were extracted, and complementary DNA (cDNA) was synthesized from RNA using RT reaction. cDNA and genomic DNA were subjected to PCR with primers complementary to the 16S rRNA sequences of Streptococcus spp. McNemar's test was used to compare the detection rate of both assays (P<0.05). Results Streptococci were detected in 28.12% (9/32) and 37.5% (12/32) of S1 samples using rRNA- and rDNA-based PCR assays, respectively. In contrast, they were detected in only 6.25% (2/32) of S2 samples using rRNA-based RT-PCR, compared to 15.62% (5/32) using rDNA-based PCR. Finally, in S3 samples, streptococci were not detected by rRNA, whereas rDNA-based PCR still detected the bacteria in 12.5% (4/32) of the samples. The total number of PCR-positive reactions in the rDNA-based PCR was higher than in the rRNA-based assay (P<0.05). Conclusions The rRNA-based RT-PCR showed a lower detection rate of streptococci when compared to the rDNA-based PCR, suggesting that the latter may have detected dead cells of streptococci in root canal samples.


Subject(s)
Humans , Streptococcus/isolation & purification , DNA, Ribosomal/isolation & purification , RNA, Ribosomal/isolation & purification , Polymerase Chain Reaction/methods , Reverse Transcriptase Polymerase Chain Reaction/methods , Dental Pulp Cavity/microbiology , Root Canal Therapy/methods , Streptococcus/genetics , DNA, Bacterial/isolation & purification , DNA, Bacterial/genetics , DNA, Ribosomal/genetics , RNA, Bacterial/isolation & purification , RNA, Bacterial/genetics , RNA, Ribosomal/genetics , Reproducibility of Results
8.
J. appl. oral sci ; 27: e20180699, 2019. graf
Article in English | LILACS, BBO | ID: biblio-1012504

ABSTRACT

Abstract Objective This study investigated the role of extracellular deoxyribonucleic acid (eDNA) on Enterococcus faecalis ( E. faecalis ) biofilm and the susceptibility of E. faecalis to sodium hypochlorite (NaOCl). Methodology E. faecalis biofilm was formed in bovine tooth specimens and the biofilm was cultured with or without deoxyribonuclease (DNase), an inhibitor of eDNA. Then, the role of eDNA in E. faecalis growth and biofilm formation was investigated using colony forming unit (CFUs) counting, eDNA level assay, crystal violet staining, confocal laser scanning microscopy, and scanning electron microscopy. The susceptibility of E. faecalis biofilm to low (0.5%) or high (5%) NaOCl concentrations was also analyzed by CFU counting. Results CFUs and biofilm formation decreased significantly with DNase treatment (p<0.05). The microstructure of DNase-treated biofilms exhibited less structured features when compared to the control. The volume of exopolysaccharides in the DNase-treated biofilm was significantly lower than that of control (p<0.05). Moreover, the CFUs, eDNA level, biofilm formation, and exopolysaccharides volume were lower when the biofilm was treated with DNase de novo when compared to when DNase was applied to matured biofilm (p<0.05). E. faecalis in the biofilm was more susceptible to NaOCl when it was cultured with DNase (p<0.05). Furthermore, 0.5% NaOCl combined with DNase treatment was as efficient as 5% NaOCl alone regarding susceptibility (p>0.05). Conclusions Inhibition of eDNA leads to decrease of E. faecalis biofilm formation and increase of susceptibility of E. faecalis to NaOCl even at low concentrations. Therefore, our results suggest that inhibition of eDNA would be beneficial in facilitating the efficacy of NaOCl and reducing its concentration.


Subject(s)
Animals , Cattle , Sodium Hypochlorite/pharmacology , DNA, Bacterial/pharmacology , Enterococcus faecalis/growth & development , Enterococcus faecalis/drug effects , Biofilms/growth & development , Biofilms/drug effects , Deoxyribonucleases/pharmacology , Polysaccharides, Bacterial/isolation & purification , Time Factors , Microscopy, Electron, Scanning , Colony Count, Microbial , Microbial Sensitivity Tests , Reproducibility of Results , Microscopy, Confocal , Dental Pulp Cavity/microbiology
9.
Braz. oral res. (Online) ; 33: e021, 2019. tab, graf
Article in English | LILACS | ID: biblio-1001593

ABSTRACT

Abstract: This study investigated the effectiveness of XP-Endo Finisher (XPF) associated with XP-Endo Shaper (XPS) or Reciproc Blue (RB) files in reducing bacterial load in oval-shaped root canals (RC) during chemomechanical preparation (CMP) using 0.9% saline solution (NaCl) or 2.5% sodium hypochlorite (NaOCl). Eighty mandibular incisors with single oval-shaped RC were contaminated with Enterococcus faecalis. The teeth were randomly assigned to eight experimental groups (n = 10) according to the CMP, as follows: G1: XPS, G2: XPS + XPF, G3: RB, and G4: RB + XPF. CMP was performed with NaCl or NaOCl. The reduction of bacterial load was assessed by colony-forming unit count before (S1) and after (S2) CMP. Data normality was verified by using Shapiro-Wilk test. ANOVA, Tukey's test, and Bonferroni post-hoc test were used at a 5% significance level. Culturable bacteria were present in all S1 samples (p>0.05). All instrumentation techniques were effective in reducing bacterial load, irrespective of the irrigating solution (p < 0.05). With the use of NaCl, RB was more effective than XPS (p = 0.035). With the use of NaOCl, XPS and RB presented similar effectiveness (p = 0.779). XPF enhanced the bacterial reduction of both systems tested (p < 0.05). The use of NaOCl improved the CMP, irrespective of the instrumentation technique used (p < 0.05). In conclusion, XPS and RB files are effective in reducing bacterial levels in oval-shaped RC. The use of XPF as a method of agitation of the irrigating solution improved the cleaning efficiency of both file systems tested. Mechanical preparation performed with saline solution decreased culturable bacteria from the root canal, but antimicrobial substances such as NaOCl should be used to achieve a significantly better disinfection.


Subject(s)
Humans , Root Canal Preparation/instrumentation , Dental Instruments , Dental Pulp Cavity/anatomy & histology , Bacterial Load , Sodium Hypochlorite/therapeutic use , Materials Testing , Gram-Positive Bacterial Infections , Enterococcus faecalis/isolation & purification , Dental Pulp Cavity/microbiology , Disinfectants/therapeutic use , Saline Solution/therapeutic use , Incisor
10.
Braz. oral res. (Online) ; 33: e039, 2019. tab, graf
Article in English | LILACS | ID: biblio-1001610

ABSTRACT

Abstract: This clinical study compared the effectiveness of two rotary systems: HyFlex CM (Coltene-Whaledent, Altstetten, Switzerland) and ProTaper Next (Dentsply Sirona, Ballaigues, Switzerland) on the removal of cultivable bacteria and endotoxins from primarily infected root canals. This study was designed as a randomized single-blinded, 2-arm, clinical trial. Twenty-four primarily infected root canals were selected and randomly divided into 2 groups: HyFlex CM (n = 12); and ProTaper Next (n = 12). Samples were collected before and after the biomechanical preparation and inoculated in specific flasks. Irrigation was performed using 2.5% sodium hypochlorite. A kinetic turbidimetric lysate assay of limulus amoebocytes was used to quantify endotoxins. Microbiological culture technique was used to determine the count of bacterial colony forming units (CFU/mL). Data collected were statistically analyzed using SigmaPlot 12.0 for Windows. The Two-Way ANOVA statistical test was performed and the level of significance was 5%. In the samples before the biomechanical preparation, cultivable bacteria and endotoxins were evidenced in 100% of the cases. The culture analysis revealed that there was no statistically significant difference in the bacterial reduction between the two instrumentation systems. Endotoxins were present in 100% of the canals after instrumentation and there was no statistical difference between the two systems in endotoxin reduction. Thus, it was concluded that both instrumentation systems were effective in reducing root canal bacteria and endotoxins with primary endodontic infection and that there was no statistical difference between them. However, no system was able to eliminate 100% of the bacteria and their by-products.


Subject(s)
Humans , Male , Female , Adult , Bacteria/isolation & purification , Lipopolysaccharides/analysis , Dental Instruments , Dental Pulp Cavity/microbiology , Materials Testing , Bacteriological Techniques , Treatment Outcome , Root Canal Preparation/instrumentation , Endotoxins/analysis , Bacterial Load , Middle Aged
11.
Braz. dent. j ; 29(5): 459-464, Sept.-Oct. 2018. tab
Article in English | LILACS | ID: biblio-974175

ABSTRACT

Abstract The aim of this study was to compare the efficacy of grape seed extract (GSE), calcium hypochlorite [Ca(ClO)2], and sodium hypochlorite (NaOCl) irrigant solutions with rotary or reciprocating instrumentation for disinfection of root canals inoculated with Enterococcus faecalis. The mesiobuccal root canals of mandibular molars were prepared and inoculated with Enterococcus faecalis for 21 days. The roots were then randomly divided into the following eight experimental groups (n=11) according to the instrumentation technique and disinfection protocol: ProTaper Next or Reciproc R25 with sodium chloride (control group), 6% NaOCl, 6% Ca(ClO)2, or 50% GSE used for irrigation during instrumentation. The antimicrobial activity was determined on the basis of a reduction in colony-forming units (CFUs) counted on bacterial samples collected before and after root canal instrumentation and expressed as a percentage of reduction. Data were evaluated by two-way ANOVA followed by Tukey HSD post-hoc tests (p<0.05). No significant differences were observed in bacterial reduction between the ProTaper Next and Reciproc R25 systems (p>0.05), regardless of the irrigant solution used. Furthermore, all active solutions (6% NaOCl, 50% GSE, and 6% Ca(ClO)2) showed similar potential to reduce bacterial counts (p>0.05) and were significantly more effective than sodium chloride (control) (p<0.05). The results suggest that the GSE and Ca(ClO)2 have potential clinical application as irrigant solutions in endodontic therapy since they present bactericidal efficacy against Enterococcus faecalis.


Resumo O objetivo deste estudo foi comparar a eficácia do extrato de semente de uva (ESU), hipoclorito de cálcio [Ca(ClO)2] e hipoclorito de sódio (NaOCl) como soluções irrigadores quando utilizadas com instrumentos reciprocantes e rotatórios para desinfecção de canais radiculares infectados com Enterococcus faecalis. Raízes mesio-vestibulares de molares inferiores foram preparados e inoculados com E. faecalis por 21 dias. As raízes foram aleatoriamente divididas em 8 grupos (n=11) de acordo com a técnica de instrumentação e protocolo de irrigação: ProTaper Next ou Reciproc R25 associados com soro fisiológico (grupo controle), Ca(ClO)2 6%, NaOCl 6% ou ESU 50%. A atividade antimicrobiana foi determinada pela redução do número de Unidades Formadoras de Colonias (UFCs) coletadas antes e após a instrumentação e expressas em porcentagens de redução. Os dados foram analisados estatisticamente pelos testes ANOVA seguido pelo teste complementar de Tukey HSD (p<0,05). Não foi encontrado diferença estatisticamente significante na redução bacteriana entre os sistemas ProTaper Next e Reciproc R25 (p>0.05), independente da solução irrigadora usada. Além disso, todas as soluções ativas (NaOCl, ESU e Ca(ClO)2) mostraram similar potencial em reduzir a quantidade de bactérias (p>0.05) e foram significativamente mais efetivas que o soro fisiológico (p<0.05). Pode-se concluir que o ESU e o Ca(ClO)2 apresentam potencial para aplicação clínica como irrigantes endodônticos uma vez que apresentaram efetividade antimicrobiana contra o E. faecalis.


Subject(s)
Humans , Root Canal Irrigants/pharmacology , Disinfection/methods , Enterococcus faecalis/drug effects , Anti-Bacterial Agents/pharmacology , Sodium Hypochlorite/pharmacology , Stem Cells , In Vitro Techniques , Calcium Compounds/pharmacology , Composite Resins/chemistry , Dental Instruments , Dental Pulp Cavity/microbiology , Grape Seed Extract/pharmacology , Molar
12.
Braz. dent. j ; 29(3): 249-253, May-June 2018. tab
Article in English | LILACS | ID: biblio-951547

ABSTRACT

Abstract The aim of this study was to evaluate the WaveOne Gold and One Shape New Generation systems regarding the bacterial removal from root canals infected with Enterococcus faecalis by comparing them to the conventional WaveOne and One Shape systems. Forty-eight distobuccal root canals of maxillary molars sterilized with ethylene oxide were infected with E. faecalis for 21 days, and then root canal initial bacterial sample was collected with paper cones and plated on M-enterococcus agar. The specimens were randomly divided into 4 groups according to the instrumentation: WaveOne Gold, One Shape New Generation, WaveOne and One Shape. After instrumentation, samples were collected with use of scraping and paper cones at immediate and 7 days after instrumentation. The bacterial reduction was calculated and then made intragroup analysis by Friedman test and intergroup analysis by Kruskal-Wallis with Dunn's post-hoc test, all at 5% significance. All techniques significantly reduced the number of bacteria in the root canal (p<0.05). WaveOne Gold and One Shape New Generation promoted higher bacterial reduction than WaveOne and One Shape systems (p<0.05), but no significant difference was found between WaveOne Gold and One Shape New Generation or between WaveOne and One Shape (p>0.05). Novel single-file systems promote better bacterial removal than the conventional single-file systems.


Resumo A proposta deste estudo foi avaliar os sistemas WaveOne Gold e One Shape New Generation em relação à remoção bacteriana de canais infectados com Enterococcus faecalis, comparando-os com seus sistemas convencionais WaveOne e One Shape. Quarenta e oito canais disto vestibulares de molares superiores esterilizados em óxido de etileno foram contaminados com E. faecalis por 21 dias, e então acoleta bacteriana inicial foi feita com cone de papel e plaqueadas em M-enterococcus agar. Os espécimes foram aleatoriamente divididos em quarto grupos de acordo com a instrumentação: WaveOne Gold, One Shape New Generation, WaveOne e One Shape. Após instrumentação, amostras foram coletadas utilizando limagem e cones de papel imediatamente e 7 dias após o preparo. A redução bacteriana foi calculada e então feita análise intra grupos com teste de Friedman, e entre grupos utilizando Kruskal-Wallis e teste de Dunn, todos a 5% de significância. Todas as técnicas reduziram significantemente o número de bactérias do canal radicular (p<0.05). WaveOne Gold e One Shape New Generation promoveram maior redução bacteriana que WaveOne e One Shape (p<0.05), mas nenhuma diferença significante foi encontrada entre WaveOne Gold e One Shape New Generation ou entre WaveOne e One Shape (p>0.05). Novos sistemas de lima-única promovem melhor remoção bacteriana que seus sistemas convencionais.


Subject(s)
Humans , Enterococcus faecalis/isolation & purification , Root Canal Preparation/instrumentation , Dental Pulp Cavity/microbiology , In Vitro Techniques , Pilot Projects , Dental Instruments/standards , Equipment Design , Bacterial Load , Maxilla , Molar/surgery
13.
Int. j. odontostomatol. (Print) ; 12(1): 113-119, Mar. 2018. tab, graf
Article in English | LILACS | ID: biblio-893310

ABSTRACT

ABSTRACT: Molecular techniques that provide valuable information about the epidemiology of oral strains. The purpose of this study was to determine the genetic relatedness of 83 Enterococcus faecalis strains isolated from treated root canals. These strains were obtained from patients who were treated for persistent endodontic infections. E. faecalis isolates were molecular typed by Pulsed Field Gel Electrophoresis using Smal. Ten clonal groups and 13 pulse types with 38.7 % similarity for the less related strains were identified. Genetic heterogeneity among strains from different patients and a high level of genetic homogeneity among intrapatient strains were observed. Therefore, restriction endonuclease fingerprinting of genomic DNA from E. faecalis strains confirmed the polyclonality of the isolates obtained from the root canals of patients diagnosed with persistent endodontic infections, compared with other reports. These results provide additional data for a better understanding of the epidemiological aspects of root canal infections by E. faecalis.


RESUMEN: Las técnicas moleculares proporcionan información valiosa sobre la epidemiología de aislados orales. El propósito de este estudio fue determinar la relación genética de 83 cepas de Enterococcus faecalis aisladas de conductos radiculares tratados. Estas cepas se obtuvieron de pacientes que fueron tratados por infecciones endodónticas persistentes. Los aislados de E. faecalis se tipificaron molecularmente por electroforesis en gel de campo pulsado usando Smal. Se identificaron diez grupos clonales y 13 pulsotipos con un 38,7 % de similitud para las cepas menos relacionadas. Se observó heterogeneidad genética entre las cepas de diferentes pacientes y un alto nivel de homogeneidad genética entre las cepas intrapacientes. Por lo tanto, la toma de huellas dactilares a traves de restricción de ADN genómico de cepas de E. faecalis confirmó la policlonalidad de los aislados obtenidos de los conductos radiculares de pacientes diagnosticados con infecciones endodónticas persistentes, en comparación con otros informes. Estos resultados proporcionan datos adicionales para una mejor comprensión de los aspectos epidemiológicos de las infecciones del conducto radicular por E. faecalis.


Subject(s)
Humans , Periapical Periodontitis/microbiology , Enterococcus faecalis/isolation & purification , Tooth Apex/microbiology , DNA, Bacterial/analysis , Bacterial Typing Techniques , Gram-Positive Bacterial Infections/microbiology , Electrophoresis, Gel, Pulsed-Field , Dental Pulp Cavity/microbiology
15.
Braz. j. microbiol ; 49(1): 184-188, Jan.-Mar. 2018. tab, graf
Article in English | LILACS | ID: biblio-889206

ABSTRACT

ABSTRACT Removal of bacterial biofilm from the root canal system is essential for the management of endodontic disease. Here we evaluated the antibacterial effect of N-acetylcysteine (NAC), a potent antioxidant and mucolytic agent, against mature multispecies endodontic biofilms consisting of Actinomyces naeslundii, Lactobacillus salivarius, Streptococcus mutans and Enterococcus faecalis on sterile human dentin blocks. The biofilms were exposed to NAC (25, 50 and 100 mg/mL), saturated calcium hydroxide or 2% chlorhexidine solution for 7 days, then examined by scanning electron microscopy. The biofilm viability was measured by viable cell counts and ATP-bioluminescence assay. NAC showed greater efficacy in biofilm cell removal and killing than the other root canal medicaments. Furthermore, 100 mg/mL NAC disrupted the mature multispecies endodontic biofilms completely. These results demonstrate the potential use of NAC in root canal treatment.


Subject(s)
Humans , Acetylcysteine/pharmacology , Streptococcus mutans/drug effects , Actinomyces/drug effects , Enterococcus faecalis/drug effects , Biofilms/drug effects , Dental Pulp Diseases/microbiology , Lactobacillus salivarius/drug effects , Anti-Bacterial Agents/pharmacology , Streptococcus mutans/physiology , Actinomyces/physiology , Calcium Hydroxide/pharmacology , Chlorhexidine/pharmacology , Enterococcus faecalis/physiology , Dental Pulp Cavity/microbiology , Microbial Viability/drug effects , Lactobacillus salivarius/physiology
16.
J. oral res. (Impresa) ; 7(1): 24-29, ene. 22, 2018. ilus, graf, tab
Article in English | LILACS | ID: biblio-1119249

ABSTRACT

The aim of this study was to observe the penetration of an aqueous solution into the root canal dentin under sonic activation and ultrasonic activation. Materials and Method: this study consisted of experimental in vitro research. In order to achieve a closed system, the apex of 45 single-rooted teeth was sealed with wax. The step-back technique was manually performed using a K50 apical master file and 3 groups were organized according to the protocol of the final irrigant activation: group I: non-activated chinese ink for 30 seconds, group II: chinese ink sonically activated with EndoActivator for 30 seconds, and group III: chinese ink ultrasonically activated with Varios 350 equipment for 30 seconds. Teeth were sectioned longitudinally, and the samples obtained were observed under a stereomicroscope at 1X magnification in order to be photographed and scanned to calculate the penetration area using the Image J software. The tinted radicular area was evaluated in relation to the total area of the root dentin. The tukey's post-hoc test and ANOVA were used for the statistical analysis (p<0.05). Results: group I and II obtained 9.13 percent and 9.42 percent penetration respectively, while in group III the highest degree of dye infiltration was achieved (13.9 percent), being statistically significant (p<0.001). Conclusions: ultrasonic activation produced a significantly higher penetration of the dye when compared to conventional activation and sonic activation.


Subject(s)
Humans , Root Canal Irrigants/administration & dosage , Root Canal Preparation/methods , Dental Pulp Cavity/microbiology , Sonication , Ultrasonics , Epidemiology, Experimental , Root Canal Preparation/instrumentation , Therapeutic Irrigation/instrumentation , Therapeutic Irrigation/methods
17.
J. appl. oral sci ; 26: e20170374, 2018. tab
Article in English | LILACS, BBO | ID: biblio-893735

ABSTRACT

Abstract Objectives To determine the concentration of calcium, iron, manganese and zinc ions after the application of chelator to Enterococcus faecalis biofilms. Material and Methods Fifty bovine maxillary central incisors were prepared and inoculated with E. faecalis for 60 days. The following were used as irrigation solutions: 17% EDTA (pH 3, 7 and 10), 2.5% sodium hypochlorite (NaOCl) combined with 17% EDTA (pH 3, 7 and 10), distilled water (pH 3, 7 and 10), and 2.5% NaOCl. Each solution was kept in the root canal for five minutes. Fifteen uncontaminated root canals were irrigated with 17% EDTA (pH 3, 7 and 10). Six teeth were used as bacterial control. The number of calcium, iron, manganese and zinc ions was determined using flame atomic absorption spectrometry. Mean ± standard deviation (SD) values were used for descriptive statistics. Results Calcium chelation using 17% EDTA at pH 7 was higher than at pH 3 and 10, regardless of whether bacterial biofilm was present. The highest concentration of iron occurred at pH 3 in the presence of bacterial biofilm. The highest concentration of manganese found was 2.5% NaOCl and 17% EDTA at pH 7 in the presence of bacterial biofilm. Zinc levels were not detectable. Conclusions The pH of chelating agents affected the removal of calcium, iron, and manganese ions. The concentration of iron ions in root canals with bacterial biofilm was higher after the use of 17% EDTA at pH 3 than after the use of the other solutions at all pH levels.


Subject(s)
Animals , Cattle , Chelating Agents/pharmacology , Enterococcus faecalis/drug effects , Biofilms/drug effects , Dental Pulp Cavity/drug effects , Dental Pulp Cavity/microbiology , Root Canal Irrigants/pharmacology , Root Canal Irrigants/chemistry , Sodium Hypochlorite/pharmacology , Sodium Hypochlorite/chemistry , Spectrophotometry, Atomic , Materials Testing , Water/chemistry , Chelating Agents/chemistry , Calcium/analysis , Edetic Acid/pharmacology , Edetic Acid/chemistry , Enterococcus faecalis/chemistry , Dental Pulp Cavity/chemistry , Hydrogen-Ion Concentration , Ions , Iron/analysis , Manganese/analysis
18.
Braz. oral res. (Online) ; 32: e120, 2018. tab, graf
Article in English | LILACS | ID: biblio-974436

ABSTRACT

Abstract The present study aims to evaluate the longitudinal effects of induced experimental infections in gnotoxenic animals on the expression of inflammatory chemokines and their receptors in periradicular tissues. The null hypothesis tested was that Enterococcus faecalis and Fusobacterium nucleatum had no effect on CCR5, CCL5, CXCL10, CCL2/MCP-1, CXCR2 and CCR1 expression. Two groups of five animals (n = 5) aged between 8 and 12 weeks were used in this study. The animals were anaesthetized, and coronary access was performed in the first molar on the right and left sides. Microorganisms were inoculated into the left molar, and the right molar was sealed without contamination to function as a control. Animals were sacrificed 7 and 14 days after infection, and periapical tissues were collected. The cytokine mRNA expression levels were assessed using real-time PCR. The chemokine mRNA expression levels demonstrated that the experimental infection was capable of inducing increased chemokine expression on day 7 compared to that on day 14, except for CCR5 and CCL5, which showed no changes. The gnotoxenic animal model proved to be effective and allowed evaluation of the immune response against a known infection. Additionally, this study demonstrates that gene expression of chemokines and their receptors against the experimental infection preferentially prevailed during the initial phase of induction of the periradicular alteration (i.e., on day 7 post-infection).


Subject(s)
Animals , Mice , Gram-Positive Bacterial Infections/immunology , Chemokines/analysis , Receptors, Chemokine/analysis , Dental Pulp Cavity/immunology , Dental Pulp Diseases/immunology , Fusobacterium Infections/immunology , Germ-Free Life , Periapical Diseases/immunology , Periapical Diseases/microbiology , Reference Values , Time Factors , Gene Expression , Chemokines/genetics , Receptors, Chemokine/genetics , Dental Pulp Cavity/microbiology , Dental Pulp Diseases/microbiology , Real-Time Polymerase Chain Reaction
19.
Braz. oral res. (Online) ; 32(supl.1): e65, 2018. tab, graf
Article in English | LILACS | ID: biblio-974475

ABSTRACT

Abstract: Chemomechanical preparation is intended to clean, disinfect, and shape the root canal. This step is of utmost importance during treatment of infected teeth with apical periodontitis, because treatment outcome depends on how effectively the clinician eliminates bacteria, their products, and necrotic tissue that would serve as substrate for bacterial regrowth. Nonetheless, curvatures and complex internal anatomical variations of the root canal system can pose a high degree of difficulty in reaching these goals. In infected teeth, bacteria may persist not only in difficult-to-reach areas such as isthmuses, ramifications, dentinal tubules, and recesses from C-shaped or oval/flattened canals, but also in areas of the main canal wall that remain untouched by instruments. If bacteria withstand chemomechanical procedures, there is an augmented risk for post-treatment apical periodontitis. This article discloses the reasons why some areas remain unprepared by instruments and discusses strategies to circumvent this issue and enhance infection control during endodontic treatment/retreatment of teeth with apical periodontitis.


Subject(s)
Humans , Root Canal Preparation/methods , Dental Pulp Cavity/microbiology , Periapical Periodontitis/therapy , Root Canal Irrigants/therapeutic use , Treatment Outcome , Root Canal Preparation/instrumentation , Dental Instruments , Dental Pulp Cavity/anatomy & histology , Dental Pulp Cavity/diagnostic imaging , X-Ray Microtomography/methods
20.
Braz. oral res. (Online) ; 32(supl.1): e69, 2018. tab, graf
Article in English | LILACS | ID: biblio-974470

ABSTRACT

Abstract: Evidence shows the polymicrobial etiology of endodontic infections, in which bacteria and their products are the main agents for the development, progression, and dissemination of apical periodontitis. Microbial factors in necrotic root canals (e.g., endotoxin) may spread into apical tissue, evoking and supporting a chronic inflammatory load. Thus, apical periodontitis is the result of the complex interplay between microbial factors and host defense against invasion of periradicular tissues. This review of the literature aims to discuss the complex network between endodontic infectious content and host immune response in apical periodontitis. A better understanding of the relationship of microbial factors with clinical symptomatology is important to establish appropriate therapeutic procedures for a more predictable outcome of endodontic treatment.


Subject(s)
Humans , Periapical Periodontitis/microbiology , Dental Pulp Cavity/microbiology , Dental Pulp Diseases/complications , Dental Pulp Diseases/microbiology , Periapical Periodontitis/pathology , Bacterial Infections/complications , Bacterial Infections/microbiology , Lipopolysaccharides/physiology , Cytokines/analysis , Cytokines/physiology , Matrix Metalloproteinases/analysis , Matrix Metalloproteinases/physiology , Dental Pulp Cavity/pathology , Dental Pulp Diseases/pathology , Endotoxins/physiology
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