Novel Antibacterial and Biocompatible Nanostructured Gels Based on One-step Synthesis as a Potential Disinfectant for Endodontic Infection Control.

AIM: The objective of this study was to develop nanostructured gels as biocompatible intracanal disinfectants by one-step microwave radiation-assisted synthesis. METHODS: Polyvinyl alcohol (PVA) and polyvinyl pyrrolidone (PVP) were used as a support network, and polyethylene glycol (PEG) was used as a reducing agent. The gels were characterized by measuring the swelling ratio (SR) and rheological properties and by scanning electron microscopy (SEM), transmission electron microscopy (TEM) and atomic force microscopy (AFM). The antibacterial effects of each gel were evaluated against the endodontic clinical strain Enterococcus faecalis. Then, the viability of the 21-day mature multispecies bacterial biofilm was assessed using confocal microscopy in an ex vivo model, where the biofilm was exposed to the mix of nanogels. The cell proliferation, viability, and morphology of human periodontal ligament (HPDL) cells were quantified using a real-time IncuCyte® S3 Live-Cell System. Viability was measured by confocal microscopy using an ex vivo model exposing a 21-day mature multispecies bacterial biofilm to the mix of nanogels. RESULTS: The antibacterial activity of the gels coincided with the superficial characterization and the solubility of the gel in the growth medium. Gels with higher viscosity (327.85-980.58 Pa s), higher dissolution (42-70%SR), and lower porosity (no porosity and 611.63 nm) showed excellent antibacterial activity against E. faecalis. Despite their physicochemical characteristics, CuNPs gels showed greater effectiveness against E. faecalis.These nanostructured gels with high PVA concentrations promote HPDL cells proliferation while still exerting antibacterial properties. Mix of nanogels showed an increase non-viable cells biomass from at of application. CONCLUSIONS: The use of biocompatible polymers influences the physicochemical, bactericidal, and cytotoxic response, making these materials potential disinfectant agents against resistant bacteria with good biocompatibility and improved HPDL cells proliferation.

Study of the Antibacterial Capacity of a Biomaterial of Zeolites Saturated with Copper Ions (Cu2+) and Supported with Copper Oxide (CuO) Nanoparticles.

In this work, copper (II) ions were saturated and copper oxide nanoparticles (CuO NPs) were supported in natural zeolite from Chile; this was achieved by making the adsorbent material come into contact with a copper ion precursor solution and using mechanical agitation, respectively. The kinetic and physicochemical process of the adsorption of copper ions in the zeolite was studied, as well as the effect of the addition of CuO NPs on the antibacterial properties. The results showed that the saturation of copper (II) ions in the zeolite is an efficient process, obtaining a 27 g L-1 concentration of copper ions in a time of 30 min. The TEM images showed that a good dispersion of the CuO NPs was obtained via mechanical stirring. The material effectively inhibited the growth of Gram-negative and Gram-positive bacteria that have shown resistance to methicillin and carbapenem. Furthermore, the zeolite saturated with copper at the same concentration had a better bactericidal effect than the zeolite supported with CuO NPs. The results suggested that the ease of processing and low cost of copper (II) ion-saturated zeolitic material could potentially be used for dental biomedical applications, either directly or as a bactericidal additive for 3D printing filaments.

Real-time evaluation of the biocompatibility of calcium silicate-based endodontic cements: An in vitro study.

INTRODUCTION: An ideal filling material should hermetically seal the communication pathways between the canal system and surrounding tissues. Therefore, during the last few years, the development of obturation materials and techniques to create optimal conditions for the proper healing of apical tissues has been a focus of interest. The effects of calcium silicate-based cements (CSCs) on periodontal ligament cells have been investigated, and promising results have been obtained. To date, there are no reports in the literature that have evaluated the biocompatibility of CSCs using a real-time live cell system. Therefore, this study aimed to evaluate the real-time biocompatibility of CSCs with human periodontal ligament cells (hPDLCs). METHODOLOGY: hPDLC were cultured with testing media of endodontic cements for 5 days: TotalFill-BC Sealer, BioRoot RCS, Tubli-Seal, AH Plus, MTA ProRoot, Biodentine, and TotalFill-BC RRM Fast Set Putty. Cell proliferation, viability, and morphology were quantified using real-time live cell microscopy with the IncuCyte S3 system. Data were analyzed using the one-way repeated measures (RM) analysis of variance multiple comparison test (p < .05). RESULTS: Compared to the control group, cell proliferation in the presence of all cements was significantly affected at 24 h (p < .05). ProRoot MTA and Biodentine lead to an increase in cell proliferation; there were no significant differences with the control group at 120 h. In contrast, Tubli-Seal and TotalFill-BC Sealer inhibited cell growth in real-time and significantly increased cell death compared to all groups. hPDLC co-cultured with sealer and repair cements showed a spindle-shaped morphology except with cements Tubli-Seal and TotalFill-BC Sealer where smaller and rounder cells were obtained. CONCLUSIONS: The biocompatibility of the endodontic repair cements performed better than the sealer cements, highlighting the cell proliferation of the ProRoot MTA and Biodentine in real-time. However, the calcium silicate-based TotalFill-BC Sealer presented a high percentage of cell death throughout the experiment similar to that obtained.

Impact of graphene oxide lateral dimensions on the properties of methacrylated gelatin nanocomposite hydrogels.

The size and shape of nanoparticles have a profound effect on the properties of nanocomposites. For instance, the lateral dimensions of graphene oxide (GO) platelets affect several properties, including their antibacterial and pharmacokinetic functions. However, the impact of lateral dimensions has been poorly studied in nanocomposites, and their effect on hydrogels is still unknown. The current study aims to determine the effect of GO lateral dimensions on the mechanical, rheological, thermal, and antibacterial properties of gelatin hydrogels. The hydrogels were fabricated via photopolymerization of methacrylated gelatin and GO derived from the oxidation of commercial graphene. The observations indicate that an increase in GO sheets improves the mechanical strength with an increase in compressive modulus and a low mechanical hysteresis (<10%). Furthermore, low mechanical energy is dissipated even after several deformation cycles. The nanocomposite hydrogels demonstrated bactericidal effects on two clinical strains with an extensively drug-resistant phenotype, primarily through contact. Additionally, an increment in lateral dimensions increased the bactericidal capacity of Gram-negative strains. Thus, the significant effect of the lateral dimensions of GO sheets on the properties of hydrogels is demonstrated.

Bioactivity analysis of calcium silicate-based sealers and repair cements on the phenotype and cytokine secretion profile of CD14+ monocytes: An ex vivo study.

AIM: This study evaluated the immune bioactivity of testing media (TM) obtained from different calcium silicate-based sealers and cements on monocyte morphology, activation, differentiation and cytokine secretion. METHODS: Blood-derived CD14+ monocytes were isolated and cultured for 5 days with 25% TM from the following calcium silicate-based materials: TotalFill BC RRM Fast-Set Putty, Biodentine, TotalFill BC Sealer and BioRoot-Root-Canal-Sealer (RCS). A resin-based endodontic cement was used as a control. The expression of surface markers such as CD86, HLA-DR, CD16, CD309 and CD209, and cytokine secretion were analysed by flow cytometry. Data were analysed using the one-way repeated measures analysis of variance (anova) multiple comparison test and a Holm-Sidak multiple comparison post-hoc test (p < .05). RESULTS: This comparative analysis revealed that monocytes co-cultured with calcium silicate-based materials showed a spindle-shaped morphology compared with the round shape observed in the control. Regarding activation markers, BioRoot-RCS and Biodentine significantly increased CD86 expression compared with the control sample, whereas no significant differences (p > .05) were observed in HLA-DR expression. In addition, no differences were observed among the differentiation markers. When the inflammatory cytokines were analysed, BioRoot-RCS increased the secretion of IL-1ß, IL-6, IL-10 and TNF-α, whereas BioRoot-RCS and Biodentine significantly decreased IL-8 production (p < .05). CONCLUSIONS: These data showed that the calcium silicate-based materials tested changed the morphology of CD14+ monocytes; however, only BioRoot-RCS and Biodentine significantly upregulated CD86. In addition, BioRoot-RCS was the sealer with the highest immunomodulatory properties for cytokine production which means that it can contribute with the in vivo healing process and regeneration of periapical lesions.

In vitro antibacterial activity of endodontic bioceramic materials against dual and multispecies aerobic-anaerobic biofilm models.

The aim of this in vitro study was to evaluate the antibacterial activity of calcium silicate repair cements and sealers against a dual-species planktonic aerobic model with different aging times and the ability to inhibit the formation of a mature 21-day-old multispecies anaerobic biofilm. The antibacterial activity of ProRoot MTA, MTA Angelus, Biodentine, BioRoot RCS and TotalFill BC sealer against a dual-species aerobic planktonic model, as well as measuring how materials were affected by aging, was evaluated using the Modified Direct Contact Test. Subsequently, the ability to inhibit the formation of a mature multispecies anaerobic biofilm was evaluated using scanning electron microscopy complemented with confocal laser scanning microscopy. Biodentine and BioRoot RCS had higher antibacterial action, and Biodentine was able to maintain its antibacterial action after a prolonged aging period in vitro. Calcium silicate repair cement MTA ProRoot and Biodentine had higher antibiofilm action.

An in vitro study on the inhibition and ultrastructural alterations of Candida albicans biofilm by zinc oxide nanowires in a PMMA matrix.

OBJECTIVES: The purpose of this study was (i) to investigate whether nanocomposite poly(methyl-methacrylate)-zinc oxide nanowires (PMMA-ZnO-NWs) have C. albicans antibiofilm activity; (ii) to evaluate the interaction between components of the nanocomposites based on PMMA-ZnO-NWs by Raman spectroscopy; and (iii) to assess ultrastructural alterations. DESIGN: Sixty-eight rectangles (17 PMMA (control) and 51 PMMA-ZnO-NWs (250, 500, 1000 ppm ZnO nanowires) were fabricated. C. albicans ATCC 10231 and a C. albicans clinical strain were tested. Adherence, biofilm formation and ultrastructural alterations were assessed by transmission electron microscopy. Raman mapping images and spectra were analyzed using main component analysis. RESULTS: Nanocomposite PMMA-ZnO-NWs inhibited the formation of C. albicans biofilms 94% at 1000 ppm and 80% at 500 ppm against both C. albicans strains. PMMA-ZnO-NWs induced ultrastructural alterations, including cell wall damage and disorganization of the cytoplasmic membrane, resulting in cell lysis. Raman spectroscopy showed new vibrational modes (300-365-485-600 cm-1) for PMMA and ZnO-NW interactions. CONCLUSIONS: PMMA-ZnO-NWs inhibited C. albicans dose-dependent biofilm formation and led to changes in the structures and cell membrane. Raman spectroscopy showed chemical interactions between ZnO-NWs and PMMA, as suggested by the appearance of new bands at 301 and 485 cm-1.

Antimicrobial Effect of Leukocyte- and Platelet-Rich Fibrin Against Aggregatibacter actinomycetemcomitans Mono-Multiserotypes Model

ABSTRACT: Leukocyte- and platelet-rich fibrin is a widely used platelet concentrate for periodontal surgery procedures. Many benefits are described regarding its use, such as antimicrobial properties. The objective of this study was to evaluate the antimicrobial effects of the different zones of this platelet concentrated against the most prevalent serotypes of Aggregatibacter actinomycetemcomitans in an in vitro mono-multiserotype model. Three patients who were treated at a School of Dentistry in the city where the researchers reside, were included. Modified direct contact method tests and results were analyzed using multivariate logistic regression analysis. In the modified direct contact method test, a decrease in bacterial count was found at time 1, but at time 2, the count increased for all serotypes and zones of L-PRF. It can be noted that the areas with more cellular content in leukocytes and platelet-rich fibrin are the areas with the most antimicrobial powe r. This platelet concentrate would have better results with serotype c. At time point 2, it is likely to act as a growth promoter of A. actinomycetemcomitans.

RESUMEN: La Fibrina rica en Leucocitos y Plaquetas es un concentrado plaquetario ampliamente utilizado en procedimientos quirúrgicos periodontales. Muchos beneficios se describen con respecto a su uso, tales como propiedades antimicrobianas. El objetivo de este estudio fue evaluar los efectos antimicrobianos de las diferentes zonas de este concentrado plaquetario frente a los serotipos más prevalentes de Aggregatibacter actinomycetemcomitans en un modelo mono-multi serotipo in vitro. Se incluyeron tres pacientes que fueron tratados en la Facultad de Odontología de la ciudad donde residen los investigadores. Se utilizó para su análisis una prueba de contacto directo modificado. En la prueba de contacto directo modificado, se encontró una disminución en el recuento bacteriano en el tiempo 1, pero en el tiempo 2, el recuento aumentó para todos los serotipos y zonas de L-PRF. Se puede observar que las áreas con mayor contenido celular en la Fibrina rica en Leucocitos y Plaquetas son las áreas con mayor poder antimicrobiano. Este concentrado de plaquetas tendría mejores resultados con el serotipo c. En el tiempo 2, es probable que actúe como un promotor del crecimiento de A. actinomycetemcomitans.

Antibacterial Activity of Copper Nanoparticles (CuNPs) against a Resistant Calcium Hydroxide Multispecies Endodontic Biofilm.

Endodontic treatment reduces the amount of bacteria by using antimicrobial agents to favor healing. However, disinfecting all of the canal system is difficult due to its anatomical complexity and may result in endodontic failure. Copper nanoparticles have antimicrobial activity against diverse microorganisms, especially to resistant strains, and offer a potential alternative for disinfection during endodontic therapy. This study evaluated the antibacterial action of copper nanoparticles (CuNPs) on an ex vivo multispecies biofilm using plaque count compared to the antibacterial activity of calcium hydroxide Ca(OH)2. There were significant differences between the Ca(OH)2 and CuNPs groups as an intracanal dressing compared with the CuNPs groups as an irrigation solution (p < 0.0001). An increase in the count of the group exposed to 7 days of Ca(OH)2 was observed compared to the group exposed to Ca(OH)2 for 1 day. These findings differ from what was observed with CuNPs in the same period of time. Antibacterial activity of CuNPs was observed on a multispecies biofilm, detecting an immediate action and over-time effect, gradually reaching their highest efficacy on day 7 after application. The latter raises the possibility of the emergence of Ca(OH)2-resistant strains and supports the use of CuNPs as alternative intracanal medication.

Antibacterial Activity of Zinc Oxide Nanoparticles in Self- Curing Acrylic Resin Against Streptococcus mutans

ABSTRACT: Antibacterial activity of zinc oxide nanoparticles in self-curing acrylic resin against Streptococcus mutans. The main objective of this study was to investigate whether nanoparticles of zinc oxide (ZnO) in self-curing acrylic resin, hav e antimicrobial properties against Streptococcus mutans, one of the microorganisms involved in the development of caries. Self- cured acrylic resin samples were prepared by incorporating ZnO nanoparticles at different concentrations based on the minimum inhibitory concentration (MIC) for Streptococcus mutans ATCC 25175. Antibacterial activity against a biofilm was evaluated in samples that were aged in artificial saliva for different times using spectral confocal laser microscopy and scanning electron microscopy. Kruskal-Wallis test using IBM SPSS Statistics version 23.0 software (SPSS Inc. ®, Chicago, IL, United States) were used, establishing the value of p <0.05 for statistical significance. The volume of the total biomass that formed in the samples aged for one day was significantly lower than the volume of the total biomass that was formed in those aged for additional days (p <0.001). Electron microscopy analysis revealed high porosity surfaces in all samples. Bacterial clusters wer e located next to large pores and irregular surfaces, while smooth surfaces had defined and linear organization cocci or simple chains. Considering the limitations of this study, the results suggest that the antibacterial activity of ZnO nanoparticles add ed to self-curing acrylic (ALIKE) is effective, mainly in fresh 1-day samples, independent of their concentration, and in samples with 16 MIC aged for 14 days, indicating it does not lose its antibacterial activity despite setting for more days. In addition, the ZnO nanoparticles added to ALIKE have the ability to inhibit the formation of biofilms, although they do not minimize the number of viable bacteria.

RESUMEN: El objetivo principal de este estudio fue investigar si nanopartículas de óxido de zinc (ZnO), incorporadas a acrílico acrilico de autocurado, tienen propiedades antimicrobianas contra Streptococcus mutans, uno de los microorganismos implicados en el desarrollo de caries. Se prepararon muestras de resina acrílica autopolimerizada mediante la incorporación de nanopartículas de ZnO a diferentes concentraciones basadas en la concentración mínima inhibitoria (MIC) para Streptococcus mutans ATCC 25175. Se evaluó la actividad antibacteriana contra una biopelícula en muestras envejecidas en saliva artificial para diferentes tiempos utilizando espectros microscopía láser confocal y microscopía electrónica de barrido. Se utilizó la prueba de Kruskal-Wallis utilizando el software IBM SPSS Statistics versión 23.0 (SPSS Inc. ®, Chicago, IL, Estados Unidos), estableciendo el valor de p <0,05 para la significancia estadística. El volumen de la biomasa total que se formó en las muestras envejecidas durante un día fue significativamente menor que el volumen de la biomasa total que se formó en las envejecidas durante días adicionales (p <0,001). El análisis de microscopía electrónica reveló superficies de alta porosidad en todas las muestras. Los cúmulos bacterianos se ubicaron junto a poros grandes y superficies irregulares, mientras que las superficies lisas tenían cocos o cadenas simples de organización lineal y definida. Considerando las limitaciones de este estudio, los resultados sugieren que la actividad antibacteriana de las nanopartículas de ZnO agregadas al acrílico autopolimerizable (ALIKE) es efectiva, principalmente en muestras frescas de 1 día, independientemente de su concentración, y en muestras con 16 MIC envejecidas para 14 días, lo que indica que no pierde su actividad antibacteriana a pesar de estar fraguada durante más días. Además, las nanopartículas de ZnO añadidas a ALIKE tienen la capacidad de inhibir la formación de biopelículas, aunque no minimizan el número de bacterias viables.

Molecular characterization and antibacterial activity of oral antibiotics and copper nanoparticles against endodontic pathogens commonly related to health care-associated infections.

OBJECTIVE: To carry out molecular characterization and determine the antibacterial activity of oral antibiotics and copper nanoparticles (Cu-NPs) against endodontic strains isolated from persistent infections. MATERIALS AND METHODS: Root canal samples from 24 teeth in different patients with persistent endodontic infections were obtained. The isolated strains were identified by biochemical tests and 16S rDNA sequencing. Genotyping was achieved by molecular methods. The antibacterial activity of antibiotics and copper nanostructures was determined by using minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) values. Furthermore, a time-kill kinetics assay was evaluated. Nonparametric tests (Kruskal-Wallis ANOVA) were performed (p value <0.05). RESULTS: Twenty-one isolated strains were identified. Six isolates of Enterococcus faecalis were grouped into two clusters of three isolates each, two of which were clones. All were clarithromycin-resistant and erythromycin. Eight Pseudomonas putida presented two clusters, two Pseudomonas spp. were not clonal, and all were resistant to the tested antibiotics except tetracycline. Two of five strains of Cutibacterium acnes were clonal, and all were resistant only to metronidazole. The lowest MIC and MBC values were obtained with Cu-NPs. Time-kill kinetics using Cu-NPs showed a significant decrease in all tested species within 4 h and reached 100% in 2 h for C. acnes. CONCLUSION: In this study, in relation to health care-associated infections, endodontic strains of each species isolated at least in one patient were polyclonal. In Pseudomonas spp., at least one clone was shared between patients. E. faecalis and C. acnes strains were susceptible to low Cu-NP concentrations, while Pseudomonas spp. strains were resistant. CLINICAL RELEVANCE: Assessing and keeping track of the susceptibility of clinical strains to antimicrobial compounds is important for the clinical outcome. Based on our results, Cu-NPs could be an alternative for endodontic treatment, in order to avoid selection of resistant strains.

Antibacterial and cytotoxic evaluation of copper and zinc oxide nanoparticles as a potential disinfectant material of connections in implant provisional abutments: An in-vitro study.

OBJECTIVE: This study evaluates the antibacterial activity against mono and multispecies bacterial models and the cytotoxic effects of zinc oxide and copper nanoparticles(ZnO-NPs/Cu-NPs) in cell cultures of human gingival fibroblasts(HGFs). DESIGN: The antibacterial activities of ZnO-NPs and Cu-NPs against 4 bacteria species were tested according to their minimum inhibitory concentrations(MICs) and against mature multispecies anaerobic model by spectral confocal laser scanning microscopy. The viabilities and cytotoxic effects of ZnO-NPs and Cu-NPs to HGFs cell cultures were tested by MTT, LDH assays, production of ROS, and the activation of caspase-3. The results were analyzed using one-way ANOVA followed by Tukey tests, considering p < 0.05 as statistically significant. RESULTS: For all strains, MICs of ZnO-NPs and Cu-NPs were in the range of 78.3 µg/mL-3906 µg/mL and 125 µg/mL-625 ug/mL, respectively. In a multispecies model, a significant decrease in the total biomass volume(µ3) was observed in response to exposure to 125 µg/mL of each NPs for which there was bactericidal activity. Significant differences were found between the volumes of viable and nonviable biomass exposed to nanostructures with Cu-NPs compared to ZnO-NPs. Both NPs induced mitochondrial dose-dependent cytotoxicity, ZnO-NPs increases LDH release and intracellular ROS generation. Cu-NPs at a concentration of 50 µg/mL induced production of cleaved caspase-3, activating the apoptotic pathway early and at low doses. CONCLUSIONS: After 24 h, ZnO-NPs are biocompatible between 78-100 µg/mL and Cu-NPs below 50 µg/mL. Antibacterial activity in a monospecies model is strain dependent, and in a multispecies model was a lower doses after 10 min of exposure.

Antibiotic resistance of Enterococcus faecalis from persistent endodontic infections / Resistencia antibiótica de Enterococcus faecalis provenientes de infecciones endodónticas persistentes

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.

Molecular Typing of Enterococcus faecalis from Persistent Endodontic Infections / Tipificación Molecular de Enterococcus faecalis Provenientes de Infecciones Endodónticas Persistentes

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.

Antibacterial activity against Streptococcus mutans and diametrical tensile strength of an interim cement modified with zinc oxide nanoparticles and terpenes: An in vitro study.

STATEMENT OF PROBLEM: Interim restorations are occasionally left in the mouth for extended periods and are susceptible to bacterial infiltration. Thus, dental interim cements with antibacterial properties are required. PURPOSE: The purpose of this in vitro study was to determine in vitro antibacterial activity against Streptococcus mutans and to compare the diametrical tensile strength (DTSs) of dental interim cement modified with zinc oxide nanoparticles (ZnO-NPs) with that of cement modified with terpenes. MATERIAL AND METHODS: Antibacterial properties of ZnO-NPs, terpenes, and dental interim cement modified with ZnO-NPs and cement modified with terpenes against S mutans were tested according to minimum inhibitory concentration (MIC) and direct contact inhibition (DCI). Tensile strength levels were evaluated using DTS. Results were analyzed using the Kolmogorov-Smirnov, ANOVA, and Tamhane tests (α=.05). RESULTS: The MICs of ZnO-NPs and terpenes against S mutans were 61.94 µg/g and 0.25% v/v, respectively. The DCI assay under the cylinders of cement (area of contact with the agar surface) revealed significant bacterial growth inhibition on Temp-Bond NE specimens with ZnO-NPs at MIC of 495.2 µg/g (8× MIC) and with terpenes at MIC 0.999% v/v (4× MIC) (P<.05). The Temp-Bond NE cement cylinder (control group) showed the lowest DTS (1.05 ±0.27 MPa) of all other test groups. In the Zn-NPs group, the greatest increase occurred in the NP8 (8× MIC; 495.2 µg/g) group with a value of 1.50 ±0.23 MPa, a significant increase in DTS compared with the control and terpene groups (P<.05). In the terpene group, the highest increase corresponded to group T2 (2× MIC; 0.4995% v/v) with a value of 1.29 ±0.18 MPa. CONCLUSIONS: The addition of terpenes and ZnO-NPs to interim cement showed antibacterial activity when in contact with S. mutans ATCC 25175. Both terpenes and ZnO-NPs antimicrobial agents increased diametral tensile strength.

Copper nanoparticles as potential antimicrobial agent in disinfecting root canals: a systematic review / Nanopartículas de cobre como potencial agente antimicrobiano en la desinfección de canales radiculares: revisión sistemática

Copper was registered as the first solid antimicrobial material. Its availability makes it an important option as an antibacterial agent. At nanoparticle size it does not exceed 100 nm, allowing close interaction with microbial membranes, enhancing its effect even more. Copper generates toxic hydroxyl radicals that damage cell membranes of Gram-negative and Gram-positive bacteria, among the latter, Enterococcus faecalis, which are present in infected radicular canals. Synthesis of metal nanoparticles with antimicrobial properties has become a viable alternative and has promising applications in the fight against pathogenic microorganisms. Furthermore, the use of some polymers to stabilize nanoparticles increases their release time and may as well decrease the risk of bacterial recolonization and biofilm formation within the ducts, enhancing the antimicrobial properties of these compounds.The aim of this article is to conduct a systematic review of the literature on antimicrobial copper nanoparticles, their current applications and their potential use in the area of oral health, specifically in the field of endodontics.

El cobre fue registrado como el primer material antimicrobiano sólido y su disponibilidad permite que constituya una importante opción como agente antibacteriano. Al tamaño de nanopartícula no supera los 100 nm, lo que permite una interacción estrecha con las membranas microbianas, potenciando aún más su efecto. Genera radicales hidroxilos tóxicos que provocan daño en la membrana celular de bacterias Gram negativas y Gram positivas, entre estas últimas, Enterococcus faecalis, presente en conductos radiculares infectados. Por lo que la síntesis de nanopartículas metálicas con propiedades antimicrobianas se ha vuelto una alternativa viable con aplicaciones prometedoras en la lucha contra microorganismos patógenos. Además, utilizando algunos polímeros para estabilizar las nanopartículas se aumenta el tiempo de liberación de éstas y se disminuye el riesgo de recolonización bacteriana y la formación de biopelículas al interior de los conductos, mejorando las propiedades antimicrobianas de estos compuestos. El propósito de este artículo es realizar una revisión sistemática de la literatura acerca de nanopartículas de cobre como antimicrobiano, sus aplicaciones actuales y su potencial uso en el área de la salud oral, específicamente en el campo de la Endodoncia.

Ex vivo effect of copper sulfate on Enterococcus faecalis in root canals / Efecto ex vivo del sulfato de cobre sobre Enterococcus faecalis en canales radiculares

The incomplete disinfection of root canal system has been reported as the main cause of post-treatment disease, due to the persistence of bacteria. For over thirty years Enterococcus faecalis has been considered the most common bacterial species isolated from persistent root canal infections, resisting antibacterial agents, such as chlorhexidine and calcium hydroxide. Several studies have indicated that copper has optimal disinfecting capacities in a hospital environment. Aimed to know the ex vivo effect of copper sulfate (CuSO4) on E. faecalis. Thirty-six extracted human tooth root canals were inoculated with E. faecalis ATCC 29212. These root canals had undergone endondontic procedures with a rotatory system. The effect of CuSO4 was determined by plate count method of E. faecalis obtained from one sample of each tooth under three incubation times (4th, 7th and 10th day). The canals medicated with CuSO4, the bacterial count decreased 6 log after 4 days and remained as such without statistically significant change until the tenth day. It is an indisputable fact of its antibacterial action. The bacterial persistence may be due to the ability of E. faecalis to remain viable in root canals up to 12 months without additional nutrients. These preliminary results couldbe used for further scientific work assessing the potential for the use of cooper in dentistry.

La desinfección incompleta del sistema de canales radiculares ha sido reportada como la principal causa de infección post tratamiento, debido a la persistencia bacteriana. Por más de 30 años, Enterococcus faecalis se ha considerado una de las especies bacterianas más comunes aisladas de infecciones persistentes del canal radicular, resistiendo a agentes antibacterianos, como clorhexidina e hidróxido de calcio. Varios estudios han indicado que el cobre tiene una capacidad de desinfección óptima en un entorno hospitalario. Con el objetivo de conocer el efecto ex vivo de sulfato de cobre (CuSO4) sobre E. faecalis. Treinta y seis canales radiculares de dientes humanos extraídos fueron inoculados con E. faecalis ATCC 29212. Estos canales radiculares fueron previamente sometidos a instrumentación con un sistema de rotación. El efecto de CuSO4 se determinó por el método de recuento en placa de E. faecalis a partir de una muestra de cada diente en tres tiempos de incubación (4, 7 y 10 días). En los canales medicados con CuSO4, el recuento de bacterias se redujo 6 log después de 4 días y se mantuvo como tal sin cambio estadísticamente significativo hasta el día 10. Esto demuestra su poder antibacteriano. La persistencia bacteriana pudo ser deberse a la capacidad de E. faecalis para seguir siendo viable en canales hasta 12 meses sin nutrientes adicionales. Estos resultados preliminares podrían ser utilizados para otros trabajos científicos que evaluen el potencial uso de cobre en odontología.

Enterococcus spp. isolated from root canals with persistent chronic apical periodontitis in a Chilean population

Abstract Aim: To isolate and identify in a Chilean population,Enterococcus spp. from root canals with persistent chronic apical periodontitis (CAP) and to investigate the potential correlation between the bacteria and the observed clinical features. Methods: Twenty patients with indication for endodontic retreatment due to persistent CAP were selected. Data from patient general health and dental clinical history were recorded. During retreatment, a microbial sample was obtained from the root canal and inoculated in a selectiveEnterococcus medium. Using bacterial cultivation methods, bacterial isolates belonging to the genusEnterococcus were identified. The relationship between the number of colony-forming units of Enterococcus spp. and patient clinical data was assessed statistically by the Pearson Chi square and Fisher exact tests. Finally, a Polymerase Chain Reaction (PCR) assay to determine the most prevalent species of Enterococcus spp. was conducted in the clinical samples, and the results were analyzed by a proportion comparison test. Results: Enterococcus spp. strains were isolated in 70% of the patients. Most of them (98.8%) accounted for Enterococcus faecalis and only 1.2% for Enterococcus faecium. A high frequency of E. faecalis was found in teeth with inadequate endodontic treatment or dental crown restorations. Conclusions: This study concluded that E. faecalis is prevalent in root canals with persistent CAP in a Chilean population. E. faecium as found in a single case with the poorest root canal filling. Further studies are still required to investigate the presence of other species, which may be linked to persistent chronic apical periodontitis.