Effect of zinc oxide-eugenol endodontic paste on planktonic aggregates and biofilm of Enterococcus faecalis - An atomic force microscopy evaluation.

OBJECTIVE: This work aimed to evaluate the in vitro effect of zinc oxide-eugenol paste (ZOE) on planktonic aggregates (EfPA) and biofilm (EfBio) of Enterococcus faecalis, focusing on their morphological aspects observed and analyzed using atomic force microscopy (AFM). DESIGN: The eugenol and paste were characterized by Gas Chromatography coupled with Mass Spectrometry (GC-MS) and Fourier Transform Infrared Spectroscopy (FTIR), respectively. The effect of ZOE on EfPA and EfBio was evaluated by a direct-contact test through colony counting and crystal violet staining protocol. AFM images of untreated and treated EfPA and EfBio growth on bovine dentin were obtained to analyze the morphological damage caused by the treatments. RESULTS: The characterization showed high purity in the eugenol composition and chemical interaction between the components of the paste. A bactericidal effect on aggregates was observed after 6 h of exposure, and on biofilm after 24 h of treatment (p < 0.001). A disruptive effect on the biofilm was also evident. AFM images revealed the formation of EfPA, with a notable presence of an exopolysaccharide matrix. After 6 h of ZOE treatment, there was a significant increase in the size and surface roughness profile of treated cells (p < 0.05). Loss of typical cell morphology was observed after 24 h. The effect on the biofilm showed a tendency towards a less condensed biofilm pattern in the treated group, with no differences in surface roughness. CONCLUSION: ZOE presents bactericidal action on EfPA and EfBio, promoting significant morphological changes after treatment, especially in the aggregates.

Amoxicillin-Resistant Streptococci Carriage in the Mouths of Children: A Systematic Review and Meta-Analysis.

Streptococcal bacteremia that occurs during invasive dental procedures can lead to infective endocarditis (IE) in children with certain heart diseases. Prior to such procedures, antibiotic prophylaxis (AP) with amoxicillin (AMPC) is recommended. However, the detection of amoxicillin-resistant strains (AMPC-RS) in the mouths of children with heart diseases raises the concern that they would be uncovered by the action of standard AP. This work carried out a systematic review and meta-analysis regarding AMPC-RS carriage in the mouths of children. We consulted databases covering studies between the years 2000 and 2021, following the PRISMA declaration. A meta-analysis was carried out to assess the prevalence of children carrying AMPC-RS in the mouths. The antimicrobial tests were carried out by microdilution (46.2% of articles), disk diffusion (38.3%), and the E-test (15.4%). Streptococcus mitis and S. sanguinis were bacteria with the most found resistance phenotype, with MIC reaching values of 128 µg/mL. Of the 13 selected articles, only 6 presented results that made it possible to calculate the prevalence of children carrying AMPC-RS in their mouths, ranging from 5.5% to 86.3%. Most of the studies were classified as high quality, and the collected data demonstrate the presence of streptococcal strains with different levels of resistance in the collected samples, such as the dental plaque. The meta-analysis pointed to evidence of AMPC-RS being carried, with a prevalence of 21.3% (I² = 0%, p = 0.705). There is an important prevalence of AMPC-RS carriage in the mouths of children. Specific attention should be directed to AP in those susceptible to IE.

Decontamination of Gutta-percha Cones employed in Endodontics.

The gutta-percha cones used in endodontic treatment are produced in aseptic conditions and their composition includes zinc oxide, which is responsible for antibacterial activity. However, there is the possibility of microbial contamination by manipulation, aerosol or during storage. Although several chemical agents have been tested for their decontamination, there is no consensus on the best disinfection protocol to be used. The aim of this study was to evaluate the decontamination of gutta-percha cones contaminated with the bacteria Enterococcus faecalis, by using chlorhexidine digluconate (CHX) and sodium hypochlorite (NaClO) at different concentrations for short exposure times. For this purpose, gutta-percha cones (size 40) were selected at random from a sealed box and immersed for 1 min in a microbial suspension. Then they were immersed in specific Petri dishes for different groups containing: CHX 2%, NaClO 1% or NaClO 2.5% for 30 s or 1 min, and subsequently placed in tubes containing BHI broth. After incubating the tubes for 48 h, it was observed that 1% and 2.5% NaClO and 2% CHX were effective for decontaminating the cones at those exposure time intervals. Microbial growth was detected in one of the replicates of the group with CHX applied for 30 s. To prevent the possibility of failures at this stage, the exposure time of gutta-percha cones to the decontaminating agent should not be reduced.

Os cones de guta-percha utilizados no tratamento endodôntico são produzidos em condições assépticas e possuem óxido de zinco em sua composição, responsável pela atividade antibac-teriana. No entanto, existe a possibilidade de contaminação microbiana por manipulação, aerossol ou seu armazenamento. Embora vários agentes químicos já tenham sido testados para sua descontaminação, não há consenso sobre o melhor protocolo de desinfecção a ser usado. Nosso objetivo foi avaliar a descontaminação de cones de guta-percha contaminados com a bactéria Enterococcus faecalis, utilizando digluconato de clorexidina (CHX) e hipoclorito de sódio (NaClO) em diferentes concentrações e tempos de exposição curtos. Para esse fim, 40 cones de guta-percha foram selecionados aleatoriamente, de uma caixa selada e imersos por 1 min em uma suspensão microbiana. Em seguida, foram imersos em placas de Petri específicas para diferentes grupos contendo: CHX 2%, NaClO 1% ou 2,5%, nos tempos de exposição de 30s e 1min e subseqüentemente imersos em tubos contendo caldo BHI. Após incubação dos tubos por 48 h, observou-se que NaClO 1% e 2,5% e CHX 2% foram eficazes para a descontaminação dos cones nesses intervalos de tempo de exposição. Em uma das réplicas do grupo com CHX aplicado por 30s foi detectado crescimento microbiano. O tempo de exposição dos cones de guta-percha ao agente de descontaminação não deve ser reduzido para evitar a possibilidade de falhas nesse estágio.

Decontamination of Gutta-percha Cones employed in Endodontics / Descontaminação de cones de guta-percha empregados em endodontia

ABSTRACT The guttapercha cones used in endodontic treatment are produced in aseptic conditions and their composition includes zinc oxide, which is responsible for antibacterial activity. However, there is the possibility of microbial contamination by manipulation, aerosol or during storage. Although several chemical agents have been tested for their decontamination, there is no consensus on the best disinfection protocol to be used. The aim of this study was to evaluate the decontamination of guttapercha cones contaminated with the bacteria Enterococcus faecalis, by using chlorhexidine digluconate (CHX) and sodium hypochlorite (NaClO) at different concentrations for short exposure times. For this purpose, guttapercha cones (size 40) were selected at random from a sealed box and immersed for 1 min in a microbial suspension. Then they were immersed in specific Petri dishes for different groups containing: CHX 2%, NaClO 1% or NaClO 2.5% for 30 s or 1 min, and subsequently placed in tubes containing BHI broth. After incubating the tubes for 48 h, it was observed that 1% and 2.5% NaClO and 2% CHX were effective for decontaminating the cones at those exposure time intervals. Microbial growth was detected in one of the replicates of the group with CHX applied for 30 s. To prevent the possibility of failures at this stage, the exposure time of guttapercha cones to the decontaminating agent should not be reduced.

RESUMO Os cones de gutapercha utilizados no tratamento endodôntico são produzidos em condições assépticas e possuem óxido de zinco em sua composição, responsável pela atividade antibac te riana. No entanto, existe a possibilidade de contaminação microbiana por manipulação, aerossol ou seu armazenamento. Embora vários agentes químicos já tenham sido testados para sua descontaminação, não há consenso sobre o melhor proto colo de desinfecção a ser usado. Nosso objetivo foi avaliar a descontaminação de cones de gutapercha contaminados com a bactéria Enterococcus faecalis, utilizando digluconato de clorexidina (CHX) e hipoclorito de sódio (NaClO) em diferentes concentrações e tempos de exposição curtos. Para esse fim, 40 cones de gutapercha foram selecionados aleatoriamente, de uma caixa selada e imersos por 1 min em uma suspensão microbiana. Em seguida, foram imersos em placas de Petri específicas para diferentes grupos contendo: CHX 2%, NaClO 1% ou 2,5%, nos tempos de exposição de 30s e 1min e subseqüentemente imersos em tubos contendo caldo BHI. Após incubação dos tubos por 48 h, observouse que NaClO 1% e 2,5% e CHX 2% foram eficazes para a descontaminação dos cones nesses intervalos de tempo de exposição. Em uma das réplicas do grupo com CHX aplicado por 30s foi detectado crescimento microbiano. O tempo de exposição dos cones de gutapercha ao agente de desconta minação não deve ser reduzido para evitar a possibilidade de falhas nesse estágio.

Quaternization of angico gum and evaluation of anti-staphylococcal effect and toxicity of their derivatives.

Modified polysaccharides have been featured as new agents against bacterial infection presenting biocompatibility in their use for medical purposes. In this work, we carried out the quaternization of Angico gum (AG). Quaternized Angico gum derivatives (QAG) were produced using a cationic moiety (3-Chloro-2-hydroxypropyl)trimethylammonium chloride onto the gum backbone. The products were characterized by FT-IR spectroscopy, Zeta potential, elemental analysis, and 1H NMR and degree of substitution (DS) was calculated. QAG were also evaluated for their anti-staphylococcal activity by determining Minimum Inhibitory and Bactericidal concentrations against pathogenic Staphylococcus spp. and by imaging using Atomic Force Microscopy. The hemolysis test and Galleria mellonella model were used to assess toxicity of gums. Our results showed that derivatives who presented highest DS (QAG-A3, 0.48 and QAG-B, 0.54) showed more effective antibacterial activity against tested bacteria, biocompatibility with erythrocytes and non-toxicity in G. mellonella model.

Anthelmintic, Antibacterial and Cytotoxicity Activity of Imidazole Alkaloids from Pilocarpus microphyllus Leaves.

Pilocarpus microphyllus Stapf ex Wardlew (Rutaceae), popularly known as jaborandi, is a plant native to the northern and northeastern macroregions of Brazil. Several alkaloids from this species have been isolated. There are few reports of antibacterial and anthelmintic activities for these compounds. In this work, we report the antibacterial and anthelmintic activity of five alkaloids found in P. microphyllus leaves, namely, pilosine, epiisopilosine, isopilosine, epiisopiloturine and macaubine. Of these, only anthelmintic activity of one of the compounds has been previously reported. Nuclear magnetic resonance, HPLC and mass spectrometry were combined and used to identify and confirm the structure of the five compounds. As regards the anthelmintic activity, the alkaloids were studied using in vitro assays to evaluate survival time and damaged teguments for Schistosoma mansoni adult worms. We found epiisopilosine to have anthelmintic activity at very low concentrations (3.125 µg mL-1 ); at this concentration, it prevented mating, oviposition, reducing motor activity and altered the tegument of these worms. In contrast, none of the alkaloids showed antibacterial activity. Additionally, alkaloids displayed no cytotoxic effect on vero cells. The potent anthelmintic activity of epiisopilosine indicates the potential of this natural compound as an antiparasitic agent. Copyright © 2017 John Wiley & Sons, Ltd.

Quaternized cashew gum: An anti-staphylococcal and biocompatible cationic polymer for biotechnological applications.

Chemical modifications to cashew gum (CG) structure have been previously reported to obtain new physicochemical characteristics, however until now there were no reports of modifications by introduction of new functional groups to add cationic character. This study presents a quaternization route for CG using a quaternary ammonium reagent. The chemical features of the quaternized cashew gum derivatives (QCG) were analyzed by: FTIR, elemental analysis, degree of substitution, Zeta potential, 1H NMR and 1H-13C correlation (HSQC). QCG were evaluated for their anti-staphylococcal activity by determining minimum inhibitory and bactericidal concentrations against pathogenic Staphylococcus spp. and by imaging using atomic force microscopy. Moreover, the mammalian cell biocompatibility were also assessed through hemolytic and cell toxicity assays. QCG presented promising antimicrobial activity against methicillin-resistant S. aureus and biocompatibility on tested cells. These results show that QCG could be a promising tool in the development of biomaterials with an anti-septic action.

Effect of neem (Azadirachta indica A. Juss) leaf extract on resistant Staphylococcus aureus biofilm formation and Schistosoma mansoni worms.

ETHNOPHARMACOLOGICAL RELEVANCE: There are ethnopharmacological reports supporting the use of neem (Azadirachta indica A. Juss) leaf against bacterial and worm infections. However there is a lack of studies about its effect on bacterial biofilm formation and Schistosoma mansoni worms. This study reports the in vitro effects of neem leaf ethanolic extract (Neem EE) on Methicillin-resistant Staphylococcus aureus (MRSA) biofilm and planktonic aggregation formation, and against S. mansoni worms. MATERIALS AND METHODS: Quantification of the Azadirachtin (AZA), thought to be one of their main compounds related to biological effects, was performed. The effect of sub-inhibitory concentrations of Neem EE on biofilm formation and planktonic aggregates of S. aureus was tested using the crystal violet dye method and atomic force microscopy (AFM) analysis, respectively. Changes in S. mansoni motor activity and death of worms were analyzed in vitro after exposition to the extract. Treated schistosomes were also examined using confocal laser scanning microscopy. RESULTS: It was observed the presence of AZA in the extract (0.14 ± 0.02 mg/L). Testing Neem EE sub-inhibitory concentrations, a significant biofilm adherence inhibition from 62.5 µg/mL for a sensitive S. aureus and 125 µg/mL for two MRSA strains was observed. AFM images revealed that as the Neem EE concentration increases (from 250 to 1000 µg/mL) decreased ability of a chosen MRSA strain to form large aggregates. In relation of anti-schistosoma assay, the extract caused 100% mortality of female worms at a concentration of 50 µg/mL at 72 h of incubation, while 300 µg/mL at 24h of incubation was required to achieve 100% mortality of male worms. The extract also caused significant motor activity reduction in S. mansoni. For instance, at 96 h of incubation with 100 µg/mL, 80% of the worms presented significant motor activity reduction. By the confocal microscopy analysis, the dorsal surface of the tegument of worms exposed to 300 µg/mL (male) and 100 µg/mL (female) of the extract showed severe morphological changes after 24h of treatment. CONCLUSIONS: Neem leaf ethanolic extract presented inhibitory effect on MRSA biofilm and planktonic aggregation formation, and anthelmintic activity against S. mansoni worms.

Collagen-based silver nanoparticles for biological applications: synthesis and characterization.

BACKGROUND: Type I collagen is an abundant natural polymer with several applications in medicine as matrix to regenerate tissues. Silver nanoparticles is an important nanotechnology material with many utilities in some areas such as medicine, biology and chemistry. The present study focused on the synthesis of silver nanoparticles (AgNPs) stabilized with type I collagen (AgNPcol) to build a nanomaterial with biological utility. Three formulations of AgNPcol were physicochemical characterized, antibacterial activity in vitro and cell viability assays were analyzed. AgNPcol was characterized by means of the following: ultraviolet-visible spectroscopy, dynamic light scattering analysis, Fourier transform infrared spectroscopy, atomic absorption analysis, transmission electron microscopy and of X-ray diffraction analysis. RESULTS: All AgNPcol showed spherical and positive zeta potential. The AgNPcol at a molar ratio of 1:6 showed better characteristics, smaller hydrodynamic diameter (64.34 ± 16.05) and polydispersity index (0.40 ± 0.05), and higher absorbance and silver reduction efficiency (0.645 mM), when compared with the particles prepared in other mixing ratios. Furthermore, these particles showed antimicrobial activity against both Staphylococcus aureus and Escherichia coli and no toxicity to the cells at the examined concentrations. CONCLUSIONS: The resulted particles exhibited favorable characteristics, including the spherical shape, diameter between 64.34 nm and 81.76 nm, positive zeta potential, antibacterial activity, and non-toxicity to the tested cells (OSCC).

Development and characterization of multilayer films of polyaniline, titanium dioxide and CTAB for potential antimicrobial applications.

Composites prepared from polyaniline (PANI) and the ceramic technology of titanium dioxide (TiO2) have been proposed, however, the interaction of these materials with greater control of molecular arrangement becomes attractive in order to achieve properties not previously described or yet the optimization of those already reported. Therefore, in this study, thin hybrid films made of polyaniline (PANI), a conductive polymer, and the technological ceramic, titanium dioxide (TiO2), were prepared by the layer-by-layer (LbL) self-assembly technique. The films were characterized by cyclic voltammetry (CV), UV-VIS spectroscopy and atomic force microscopy (AFM). Aiming to improve the dispersion of the ceramic in the polymer matrix, the commercial surfactant, cetyl trimethylammonium bromide (CTAB), was used in the formation of the films. The best condition of deposition was found showing synergic interactions between the conjugated materials. The antibacterial activity of the PANI(TiO2)/CTAB films was studied and the obtained results suggest their use as antimicrobial coatings.

Identification of Phenolic Compounds and Evaluation of Antioxidant and Antimicrobial Properties of Euphorbia Tirucalli L.

Bioactive compounds extracted from natural sources can benefit human health. The aim of this work was to determine total phenolic content and antioxidant activity in extracts of Euphorbia tirucalli L. followed by identification and quantification of the phenolic compounds, as well as their antibacterial activities. Antioxidant activities were determined by DPPH and ABTS(•+) assay. Identification of phenolic compounds was performed using high-performance liquid chromatography (HPLC), and antimicrobial activities were verified by agar dilution methods and MIC values. Total phenolic content ranged from 7.73 to 30.54 mg/100 g gallic acid equivalent. Extracts from dry plants showed higher antioxidant activities than those from fresh ones. The DPPH EC50 values were approximately 12.15 µg/mL and 16.59 µg/mL, respectively. Antioxidant activity measured by the ABTS method yielded values higher than 718.99 µM trolox/g for dry plants, while by the Rancimat(®) system yielded protection factors exceeding 1 for all extracts, comparable to synthetic BHT. Ferulic acid was the principal phenolic compound identified and quantified through HPLC-UV in all extracts. The extracts proved effective inhibitory potential for Staphylococcus epidermidis and Staphylococcus aureus. These results showed that extracts of Euphorbia tirucalli L. have excellent antioxidant capacity and moderate antimicrobial activity. These can be attributed to the high concentration of ferulic acid.

Development and antibacterial activity of cashew gum-based silver nanoparticles.

The present study describes the development of a green synthesis of silver nanoparticles reduced and stabilized by exuded gum from Anacardium occidentale L. and evaluates in vitro their antibacterial and cytotoxic activities. Characterization of cashew gum-based silver nanoparticles (AgNPs) was carried out based on UV-Vis spectroscopy, transmission electron microscopy and dynamic light scattering analysis which revealed that the synthesized silver nanoparticles were spherical in shape, measuring about 4 nm in size with a uniform dispersal. AgNPs presented antibacterial activity, especially against Gram-negative bacteria, in concentrations where no significant cytotoxicity was observed.