In vitro bacterial adherence on teeth submitted to whitening with musa paradisiaca / Adherencia bacteriana in vitro en dientes sometidos a blanqueamiento con musa paradisiaca

Objetive: To compare in vitro bacterial adherence on teeth submitted to whitening with 50% ethanolic extract of Musa paradisiaca and 35% hydrogen peroxide. Material and Methods: The study was experimental and used 18 premolars that were grouped into: G1 (control), G2 (50% ethanol extract of Musa paradisiaca) and G3 (35% hydrogen peroxide). The teeth were then exposed to a Streptococcus mutans culture for 24 hours, followed by centrifugation in thioglycolate broth. A culture on trypticase soy agar was done with a 1 in 100 dilution, and after 48 hours colony forming units (CFU) were counted. Statistical analysis was performed using the ANOVA test, complemented by the Bonferroni post-hoc. Results: Bacterial adherence was 77x105 CFU/ml in Group 3 using 35% hydrogen peroxide, 40x105 CFU/ml in Group 2 using 50% ethanol extract of Musa paradisiaca, and 89x104 CFU/ml in Group 1 (control). The difference between the three groups was significant (p=0.000). Conclusion: Both whitening methods cause bacterial adherence to the tooth surface, although to a lower degree with Musa paradisiaca.eses.

Objetivo: Comparar la adherencia bacteriana in vitro en dientes sometidos a blanqueamiento con extracto etanólico de Musa paradisiaca al 50% y con peróxido de hidrógeno al 35%. Material y Métodos: Comparar la adherencia bacteriana in vitro en dientes sometidos a blanqueamiento con extracto etanólico de Musa paradisiaca al 50% y con peróxido de hidrógeno al 35%.Resultados: La adherencia bacteriana fue de 77x105 UFC/ml con el peróxido de hidrógeno al 35%, de 40x105 UFC/ml con el extracto etanólico de Musa paradisiaca al 50% y de 89x104 UFC/ml con el control. La diferencia fue significativa entre los tres grupos (p=0.000). Conclusión: Ambos métodos de blanqueamiento causan adherencia bacteriana en la superficie dental, siendo menor con Musa paradisiaca.

Influence of EDTA and its Association with Benzalkonium Chloride on Enterococcus faecalis Adhesion to Dentin / Influencia del EDTA y su Asociación con Cloruro de Benzalconio en la Adhesión de Enterococcus faecalis a la Dentina

ABSTRACT: The aim of this in vitro study was to investigate the influence of ethylenediaminetetraacetic acid (EDTA) associated with the benzalkonium chloride (BAK) on the adhesion and formation of Enterococcus faecalis biofilms attached to coated dentin. Discs standard bovine dentin blocks were treated with the coating materials evaluated: Saline solution (control), 17 % EDTA, 17 % EDTA associated with 1 % BAK for 5 minutes and subsequently washed with saline solution. Afterwards, biofilms of E. faecalis (ATCC 29212) were grown on the surface of coated dentin blocks for time intervals of 1 hour and 7 days (n = 20) and were subsequently washed with phosphate-buffered saline (PBS). Bacterial viability and total biovolume were analyzed by confocal laser scanning microscopy (CLSM) using the Live/Dead technique. Nonparametric Kruskal-Wallis followed by Dunn tests were used to determine statistical differences (a = 5 %). The 17 % EDTA + 1 % BAK group showed significantly lower biovolume and bacterial viability values at the end of 1 hour (p < 0.05). After 7 days of contamination, the 17 % EDTA and 17 % EDTA + 1 % BAK groups showed similar results that differed statistically from those of the control group (p < 0.05). The saline solution group showed higher values. The use of BAK associated with EDTA on dentin blocks surfaces before exposure to contamination was able to interfere in the adhesion of E. faecalis to dentin. Also, dentin treatment by BAK associated with a chelating agent influences the secondary biofilm formation, which could have important effects on the long-term success of root canal treatment.

RESUMEN: El objetivo del estudio consistió en investigar in vitro, la influencia del ácido etilendiamino-tetraacético (EDTA) con cloruro de benzalconio (BAK) en la adhesión y formación de biopelículas de Enterococcus faecalis a la dentina. Discos de dentina bovina fueron tratadas con solución salina (control), 17 % de EDTA, 17% de EDTA asociado con 1 % de BAK durante 5 minutos y lavadas con solución salina. Las biopelículas de E. faecalis (ATCC 29212) se cultivaron sobre los discos de dentina durante intervalos de tiempo de 1 hora y 7 días (n = 20), lavados con solución salina tamponada con fosfato (PBS). La viabilidad bacteriana y el biovolumen total se analizaron mediante microscopía de barrido por láser (CLSM) utilizando la técnica Live / Dead. Se realizó prueba no paramétrica de Kruskal-Wallis, seguida por Dunn con una diferencia estadística (a = 5 %). El grupo de 17 % EDTA + 1 % BAK mostró valores significativamente menores de biovolumen y viabilidad bacteriana al final de 1 hora (p < 0,05). Después de 7 días de contaminación, los grupos de 17 % EDTA y 17 % EDTA + 1 % BAK mostraron resultados similares que diferían estadísticamente del grupo control (p < 0,05). La solución salina mostró valores más altos. La asociación de BAK con EDTA antes de la contaminación interfirió en la adhesión de E. faecalis. Además, el tratamiento de la dentina por BAK asociado con EDTA influye en la formación de biopelículas secundarias, lo que podría tener efectos importantes sobre el éxito a largo plazo del tratamiento del conducto radicular.

Adhesión Bacteriana en Hilo de Sutura de Seda Negra y Nylon Post Extracción Dental Simple: Ensayo Clínico Controlado / Bacterial Adhesion in Black Silk Suture Thread and Nylon Post Simple Dental Extraction: Controlled Clinical Trial

RESUMEN: El objetivo de este estudio fue comparar los hilos de sutura de seda negra y la sutura nylon con respecto a la adhesión bacteriana post extracción dental simple de piezas posteriores. El presente trabajo se desarrolló en la Clínica Odontológica y el Laboratorio de Microbiología de la Unidad de Ciencias Biomédicas y Biotecnología de la Universidad Privada Antenor Orrego (Trujillo, Perú), entre agosto y setiembre de 2014. Se trabajó con 19 unidades de muestreo para hilo de seda negra y 19 para nylon. El procedimiento de exodoncia para todos los pacientes fue estandarizado. Luego de la extracción, se procedió a suturar la herida quirúrgica, colocando dos puntos circunferenciales, uno con cada tipo de hilo. A los 7 días, los hilos fueron retirados y sometidos a análisis microbiológico. La adhesión bacteriana de los hilos de sutura seda negra y nylon fue comparada empleando el test de Wilcoxon. Además la adhesión bacteriana de cada hilo fue comparada con la adhesión en hilos estériles (control) empleando el test de Mann-Whitney para una p<0,005. Al comparar la adhesión bacteriana de ambos hilos, se encontró diferencia entre ellos (p=0,027), correspondiendo mayor adhesión al hilo de nylon. La adhesión bacteriana fue mayor en el hilo de sutura de nylon comparado con el hilo de sutura de seda negra.

ABSTRACT: The objective of the study was to compare black silk suture threads and nylon suture with respect to bacterial adhesion after simple dental extraction of posterior pieces. The present study was developed in the Clínica Estomatológica and the Laboratorio de Microbiología of the Unidad de Ciencias Biomédicas y Biotecnología de la Universidad Privada Antenor Orrego (Trujillo, Peru), between August and September 2014. We worked with 19 units sampling for black silk thread and 19 for nylon. The procedure of exodontia for all patients was standardized. After the extraction, the surgical wound was sutured, placing two circumferential points, one with each type of thread. After 7 days, the threads were removed and subjected to microbiological analysis. The bacterial adhesion of the black silk and nylon suture strands was compared using the Wilcoxon test. In addition, the bacterial adhesion of each strand was compared with the adhesion in sterile strands (control) using the Mann-Whitney test for p <0.005. When comparing the bacterial adhesion of both threads, a difference was found between them (p = 0.027), corresponding greater adhesion to the nylon thread. Bacterial adhesion was higher in nylon suture compared to black silk suture.

Influence of the incorporation of triclosan methacrylate on the physical properties and antibacterial activity of resin composite

Abstract The incorporation of antimicrobials in the composites as an attempt to reduce bacterial adhesion without jeopardizing mechanical properties is a challenge for Dentistry. Objective: To evaluate the bacterial adhesion and physical properties of a composite containing the methacrylate triclosan- derivative monomer (TM). Methodology: TM was synthesized and added to an experimental composite. Samples were divided into two groups: Control and TM (13.4 wt%). Antibacterial Activity: Three specimens of each material were prepared and placed on bacterial suspensions of Streptococcus mutans for 1, 5 and 10 days. After these periods the counting of the colonies (log10) was performed. Assays was performed in triplicate. Physical Properties: Three-body Abrasion (TBA): Ten specimens of each material were prepared and stored at 37°C/24 h. The surface roughness (Ra) and hardness (KHN) were analyzed. Next, the specimens were submitted to abrasive wear (30,000 cycles) and re-evaluated for Ra and KHN; Sorption/solubility (SS): cylindrical specimens (n=10) were prepared and weighted. The specimens were immersed in deionized water for 7 days at 37°C and then their weight was verified again. SS were calculated using accepted formulas; Diametral tensile strength (DTS): specimens (n=10) underwent test performed in an Instron universal testing machine at a crosshead speed of 1 mm/min. Data were submitted to appropriate statistical tests according to data distribution and assay (p<0.05). Results: Bacterial Adhesion: TM showed a significant reduction on biofilm accumulation in the evaluated periods: 1 day (1.537±0.146); 5 days (2.183±0.138) and 10 days (4.469±0.155) when compared with Control: 1 day (4.954±0.249); 5 days (5.498±0.257) and 10 days (6.306±0.287). Physical Properties: For TBA, SS and DTS no significant difference was found between groups Control and TM. The incorporation of methacrylate triclosan-based monomer in the experimental composite reduce bacterial adhesion of S. mutans and did not affect important polymer properties.

In vitro antimicrobial effect of bioadhesive oral membrane with chlorhexidine gel

Abstract This study aimed to evaluate in vitro the antimicrobial effect of a bioadhesive chitosan-based oral membrane with chlorhexidine for local treatment of infections in the oral tissues. Five oral membranes of different compositions were tested: 5% chitosan (G1); 5% chitosan ± 0.2% chlorhexidine (G2), 5% chitosan ± 0.6% chlorhexidine (G3), 5% chitosan ± 1.0% chlorhexidine (G4), and 5% chitosan ± 2.0% chlorhexidine (G5). Also, five gel types were tested according to the following compositions: 5% chitosan gel (G6), 0.2% chlorhexidine gel (G7), 2.0% chlorhexidine gel (G8), 5% chitosan gel ± 0.2% chlorhexidine gel (G9), and 5% chitosan gel ± 2.0% chlorhexidine gel (G10). The antimicrobial action of the samples was tested against Candida albicans and Streptococcus mutans through antibiogram by measuring the inhibition halos. Data were statistically analyzed by Kruskal-Wallis and one-way ANOVA followed by Tukey test (p<0.05). The 2.0% chlorhexidine membrane (G5) and the disks containing 2.0% chlorhexidine gel (G8) showed the greatest inhibition halos for both microorganisms, with statistically significant difference when compared to others tested groups (p=0.008) only for Candida albicans inhibitions results. All the other formulations of membranes and gels showed inhibition halos, but without statistically significant difference. The bioadhesive chitosan-based oral membrane with 2% chlorhexidine and 2% chlorhexidine gel were the most effective in inhibiting the tested microorganisms.

Resumo O objetivo deste estudo foi avaliar in vitro o efeito antimicrobiano de uma bandagem oral bioadesiva de quitosana com clorexidina para o tratamento de infecções dos tecidos orais. Cinco bandagens de diferentes composições foram testadas: Quitosana 5% (G1); Quitosana 5% ± clorexidina a 0,2% (G2), Quitosana 5% ± clorexidina a 0,6% (G3), Quitosana 5% ± clorexidina a 1,0% (G4) e Quitosana 5% ± clorexidina a 2,0% (G5). Foram testados também 5 tipos de géis nas seguintes composições: Gel de Quitosana 5% (G6), Gel de clorexidina a 0,2% (G7), Gel de clorexidina a 2,0% (G8), Gel de Quitosana 5% ± clorexidina a 0,2% (G9) e Gel de Quitosana 5% ± clorexidina a 2,0% (G10). A ação antimicrobiana das amostras foi testada contra Candida albicans e Streptococcus mutans por meio do antibiograma, medindo o halo de inibição. Os dados foram analisados pelo teste de Kruskal-Wallis e ANOVA a um critério seguido pelo teste de Tukey (p<0,05). A membrana com 2,0% de clorexidina (G5) e os discos contendo gel com 2,0% de clorexidina (G8) apresentaram os maiores halos de inibição para os dois microrganismos, com diferença estatisticamente significativa em relação aos demais grupos testados (p=0,008) apenas nos resultados de inibição de C. albicans. Todas as outras formulações de membranas e géis apresentaram halo de inibição, mas sem diferença estatisticamente significativa. A bandagem oral bioadesiva de quitosana com gel de 2% de clorexidina foi a mais efetiva em inibir os microrganismos testados.

Biofilm formation by Staphylococcus aureus and Salmonella spp. under mono and dual-species conditions and their sensitivity to cetrimonium bromide, peracetic acid and sodium hypochlorite

Abstract The aim of this study was evaluated the biofilm formation by Staphylococcus aureus 4E and Salmonella spp. under mono and dual-species biofilms, onto stainless steel 316 (SS) and polypropylene B (PP), and their sensitivity to cetrimonium bromide, peracetic acid and sodium hypochlorite. The biofilms were developed by immersion of the surfaces in TSB by 10 d at 37 °C. The results showed that in monospecies biofilms the type of surface not affected the cellular density (p > 0.05). However, in dual-species biofilms on PP the adhesion of Salmonella spp. was favored, 7.61 ± 0.13 Log10 CFU/cm2, compared with monospecies biofilms onto the same surface, 5.91 ± 0.44 Log10 CFU/cm2 (p < 0.05). The mono and dual-species biofilms were subjected to disinfection treatments; and the most effective disinfectant was peracetic acid (3500 ppm), reducing by more than 5 Log10 CFU/cm2, while the least effective was cetrimonium bromide. In addition, S. aureus 4E and Salmonella spp. were more resistant to the disinfectants in mono than in dual-species biofilms (p < 0.05). Therefore, the interspecies interactions between S. aureus 4E and Salmonella spp. had a negative effect on the antimicrobial resistance of each microorganism, compared with the monospecies biofilms.

Evaluation antibacterial and antibiofilm activity of the antimicrobial peptide P34 against Staphylococcus aureus and Enterococcus faecalis

ABSTRACT The adhesion ability of bacteria to abiotic surfaces has important implications in food industries, because these organisms can survive for long periods through the biofilm formation. They can be transferred from one place to another in the industry causing contamination of the food processing environment. In this study, the antibacterial and antibiofilm activities of the antimicrobial peptide P34, characterized as a bacteriocin-like substance (BLS P34) were tested against planktonic and sessile cells of Staphylococcus aureus and Enterococcus faecalis isolated from foods. The BLS P34 showed inhibitory effect against all planktonic cells of E. faecalis. The inhibition of biofilm formation and the eradication of pre-formed biofilm were evaluated with the crystal violet assay and with the reduction of 3-bromide [4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium. The BLS P34 promoted a reduction of percentage of adhered microbial cells on the surface, not being able to perform the complete elimination of biofilm formation. The metabolic activity of S. aureus biofilms decreased considerably between 41-95%. However, E. faecalis cells showed up metabolically stimulated. The BLS P34 has the potential antibiofilm for the species S. aureus. Studies suggest more detailed approaches to a better understanding of the interactions between the antimicrobial and bacterial cells within the biofilm structure.

Bacterial adhesion on conventional and self-ligating metallic brackets after surface treatment with plasma-polymerized hexamethyldisiloxane

ABSTRACT Introduction: Plasma-polymerized film deposition was created to modify metallic orthodontic brackets surface properties in order to inhibit bacterial adhesion. Methods: Hexamethyldisiloxane (HMDSO) polymer films were deposited on conventional (n = 10) and self-ligating (n = 10) stainless steel orthodontic brackets using the Plasma-Enhanced Chemical Vapor Deposition (PECVD) radio frequency technique. The samples were divided into two groups according to the kind of bracket and two subgroups after surface treatment. Scanning Electron Microscopy (SEM) analysis was performed to assess the presence of bacterial adhesion over samples surfaces (slot and wings region) and film layer integrity. Surface roughness was assessed by Confocal Interferometry (CI) and surface wettability, by goniometry. For bacterial adhesion analysis, samples were exposed for 72 hours to a Streptococcus mutans solution for biofilm formation. The values obtained for surface roughness were analyzed using the Mann-Whitney test while biofilm adhesion were assessed by Kruskal-Wallis and SNK test. Results: Significant statistical differences (p< 0.05) for surface roughness and bacterial adhesion reduction were observed on conventional brackets after surface treatment and between conventional and self-ligating brackets; no significant statistical differences were observed between self-ligating groups (p> 0.05). Conclusion: Plasma-polymerized film deposition was only effective on reducing surface roughness and bacterial adhesion in conventional brackets. It was also noted that conventional brackets showed lower biofilm adhesion than self-ligating brackets despite the absence of film.

RESUMO Introdução: a deposição de filme de polímero a plasma foi criada para modificar as propriedades de superfície dos braquetes ortodônticos metálicos, com o intuito de inibir a adesão bacteriana. Métodos: filmes finos de polímero de hexametildisiloxano (HMDSO) foram depositados em braquetes ortodônticos de aço inoxidável convencionais (n = 10) e autoligáveis (n = 10), utilizando a técnica de radiofrequência PECVD (Plasma-Enhanced Chemical Vapor Deposition). As amostras foram divididas em dois grupos, de acordo com o tipo de braquete, e dois subgrupos após o tratamento de superfície. A microscopia eletrônica de varredura (MEV) foi realizada para avaliar a presença de adesão bacteriana sobre as superfícies das amostras (região de ranhura horizontal e aletas) e a integridade da camada de filme. A Interferometria Confocal (CI) avaliou a rugosidade, e a molhabilidade superficial foi avaliada por goniometria. Para análise de adesão bacteriana, as amostras foram expostas durante 72 horas a uma solução de Streptococcus mutans, para formação de biofilme. Os valores obtidos para a rugosidade da superfície foram analisados pelo teste de Mann-Whitney, enquanto a adesão do biofilme foi avaliada pelos testes de Kruskal-Wallis e SNK. Resultados: observaram-se diferenças estatisticamente significativas (p <0,05) para a rugosidade superficial e redução da adesão bacteriana em braquetes convencionais após o tratamento da superfície, e entre braquetes convencionais e autoligáveis. Não foram observadas diferenças estatísticas significativas entre os grupos autoligáveis (p> 0,05). Conclusão: a deposição de polímero a plasma só foi efetiva na redução da rugosidade superficial e adesão bacteriana em braquetes convencionais. Observou-se, também, que os braquetes convencionais apresentaram menor adesão ao biofilme do que os braquetes autoligáveis, apesar da ausência de filme.

Cymbopogon citratus essential oil: effect on polymicrobial caries-related biofilm with low cytotoxicity

Abstract The objective of this study was to evaluate the effects of Cymbopogon citratus essential oil and its main compound (citral) against primary dental colonizers and caries-related species. Chemical characterization of the essential oil was performed by gas chromatography/mass spectroscopy (GC/MS), and the main compound was determined. Antimicrobial activity was tested against Actinomyces naeslundii, Lactobacillus acidophilus, S. gordonii, S. mitis, S. mutans, S. sanguinis and S. sobrinus. Minimum inhibitory and bactericide concentrations were determined by broth microdilution assay for streptococci and lactobacilli reference, and for clinical strains. The effect of the essential oil on bacterial adhesion and biofilm formation/disruption was investigated. Negative (without treatment) and positive controls (chlorhexidine) were used. The effect of citral on preformed biofilm was also tested using the same methodology. Monospecies and microcosm biofilms were tested. ANOVA or Kruskal-Wallis tests were used (α=0.05). Cytotoxicity of the essential oil to human keratinocytes was performed by MTT assay. GC/MS demonstrated one major component (citral). The essential oil showed an inhibitory effect on all tested bacterial species, including S. mutans and L. acidophilus. Essential oil of C. citratus (10X MIC) reduced the number of viable cells of lactobacilli and streptococci biofilms (p < 0.05). The essential oil inhibited adhesion of caries-related polymicrobial biofilm to dental enamel (p < 0.01). Citral significantly reduced the number of viable cells of streptococci biofilm (p < 0.001). The essential oil showed low cytotoxicity to human keratinocytes. Based on these findings, this study can contribute to the development of new formulations for products like mouthwash, against dental biofilms.

Efecto del metronidazol suplementado con hidroquinona en la adherencia de Lactobacillus acidophilus en células vaginales ovinas / Effect of metronidazole supplemented with hydroquinone on the adhesion of Lactobacillus acidophilus in ovine vaginal cells

This work demonstrates that the addition of metronidazole together with a ubiquitous quinone compound reduces adherence of Lactobacillus acidophilus to ovine vaginal cells. Spectrophotometric and voltammetric studies have shown that neoformed compounds were observed in these systems; there were also changes in their electroactive composition, and the oxidant status had a significantly higher value compared to the control (p < 0.05). Based on reduction potential (E; mV), the distribution of electroactive compound concentrations suggests that the compounds with low reduction potential induce this behavior, which would indicate that the addition of metronidazole with a ubiquitous quinone compound to the vaginal system might increase the reductive capacity of these systems. This work shows that the study of behavior and fluctuations of the redox compounds that compose the vaginal environment, in terms of concentration and species of redox molecules, must be hierarchized in order to better understand the early stages of colonization by microorganisms.

Este trabajo demuestra que la incorporación de metronidazol conjuntamente con un compuesto quinónico ubicuo disminuye la adherencia de Lactobacillus acidophilus a células vaginales ovinas. Los estudios espectrofotométricos y voltamétricos mostraron que en estos sistemas aparecieron compuestos neoformados y que hubo modificaciones en la composición electroactiva; asimismo, el estatus oxidante presentó un valor significativamente superior con respecto al control (p < 0,05). Según los potenciales de reducción (E; mV), la distribución de las concentraciones de los compuestos electroactivos muestra que los compuestos con bajos potenciales de reducción inducen este comportamiento. Esto indicaría que la incorporación de esta mezcla al sistema vaginal aumentaría su capacidad reductora. El trabajo muestra que el estudio del comportamiento y las fluctuaciones de los compuestos redox que componen el ambiente vaginal, en términos de concentración y especies moleculares, debe ser jerarquizado para comprender mejor las primeras etapas de la colonización de este ambiente por parte de los microorganismos.

Influence of iron deprivation on virulence traits of mycobacteria

ABSTRACT Novel strategies to combat the ever increasing burden of drug resistance in Mycobacterium tuberculosis (MTB) causing tuberculosis (TB) remains a global concern. The ability of MTB to sense and adapt to restricted iron conditions in the hostile environment is essential for their survival and confers the basis of their success as dreadful pathogen. The striking and clinically relevant virulence trait of MTB is its ability to form biofilms and adhere to the host cells. The present study elucidated the effect of iron deprivation on biofilm formation and cell adherence of Mycobacterium smegmatis, a non-pathogenic surrogate of MTB. Firstly, we showed that iron deprivation leads to enhanced cell sedimentation rate and altered colony morphology depicting alterations in cell surface envelope properties. We explored that biofilm formation and cell adherence to polystyrene surface as well as human oral epithelial cells were considerably reduced under iron deprivation both in presence of 2,2 BP (iron chelator) and siderophore mutant Δ011-14 strain. We further investigated that the potency of three first line anti-TB drugs (Isoniazid, Ethambutol, Rifampicin) to inhibit both biofilm formation and cell adhesion were enhanced under iron deprivation in contrast to the drugs when tested alone. Taken together, by virtue of the indispensability of iron for functional virulence traits in mycobacteria, iron deprivation strategies could be further exploited against this notorious human pathogen to explore novel drug targets.

Diseño y evaluación de las propiedades antibacterianas de un prototipo de molde de audífono que incorpora cobre en su manufactura / Design and evaluation of antibacterial properties of a prototype of earmold with copper incorporated in the manufacture

Introducción: La otitis externa infecciosa es una consulta otorrinolaringológica frecuente en usuarios de audífonos. Las propiedades antibacterianas descritas del cobre, han motivado el desarrollo de aplicaciones clínicas de este metal. Objetivo: Evaluar la capacidad antibacteriana de moldes de audífonos manufacturados con polímeros sensible a luz UV, silicona y acrílico que incorporan cobre metálico, en un sistema experimental in vitro. Material y método: Se diseñaron moldes de audífonos con y sin cobre, que fueron inoculados con distintas concentraciones de microorganismos (S aureus y P aeruginosa), para luego determinar el porcentaje de adherencia bacteriana a distintos tiempos de contacto (4, 8 y12 horas). Resultados: Existió reducción significativa en la adhesión bacteriana a los moldes con cobre respecto a aquellos sin cobre, independiente del material, del tipo de microorganismo y del inóculo bacteriano. Discusión: La disminución en la adherencia bacteriana en los prototipos con cobre, puede atribuirse a su actividad inhibitoria sobre los microorganismos en función de su concentración y el tiempo de contacto, ejerciendo su efecto por difusión a través de los distintos materiales. Conclusión: Con estos resultados, se hace necesario el desarrollo de estudios clínicos enfocados en comprobar si el uso de audífonos con cobre disminuyen las otitis externas de origen infeccioso.

Introduction: External otitis of infectious etiology among hearing aid users is a common motive of otolaryngology consultation. Antibacterial properties described copper, they have motivated the development of clinical applications of this metal. Aim: Evaluate the antibacterial capacity of copper-based ear molds manufactured with different materials such UV sensitive polymers, silicone and acrylic incorporating metallic copper, in an experimental system in vitro. Material and method: Ear molds with and without copper, were inoculated with different concentrations of microorganisms (S aureus and P aeruginosa) and determine the percentage of bacterial adherence to different contact times (4-8 and 12 hours). Results: There was significant reduction in bacterial adhesion to copper molds than those without copper, independent of the material, the type of microorganism and the bacterial inoculum. Discussion: The decrease in bacterial adherence on prototypes with copper, can be attributed to inhibitory activity on microorganisms depending on their concentration and contact time, exerting its effect by diffusion through the various materials. Conclusion: With these results, is necessary the development of clinical studies focused on checking whether the use of hearing aids with copper decreases external otitis of infectious origin.

Antiseptics and microcosm biofilm formation on titanium surfaces

Abstract Oral rehabilitation with osseointegrated implants is a way to restore esthetics and masticatory function in edentulous patients, but bacterial colonization around the implants may lead to mucositis or peri-implantitis and consequent implant loss. Peri-implantitis is the main complication of oral rehabilitation with dental implants and, therefore, it is necessary to take into account the potential effects of antiseptics such as chlorhexidine (CHX), chloramine T (CHT), triclosan (TRI), and essential oils (EO) on bacterial adhesion and on biofilm formation. To assess the action of these substances, we used the microcosm technique, in which the oral environment and periodontal conditions are simulated in vitro on titanium discs with different surface treatments (smooth surface - SS, acid-etched smooth surface - AESS, sand-blasted surface - SBS, and sand-blasted and acid-etched surface - SBAES). Roughness measurements yielded the following results: SS: 0.47 µm, AESS: 0.43 µm, SB: 0.79 µm, and SBAES: 0.72 µm. There was statistical difference only between SBS and AESS. There was no statistical difference among antiseptic treatments. However, EO and CHT showed lower bacterial counts compared with the saline solution treatment (control group). Thus, the current gold standard (CHX) did not outperform CHT and EO, which were efficient in reducing the biofilm biomass compared with saline solution.

Effects of penicillin and erythromycin on adherence of invasive and noninvasive isolates of Streptococcus pyogenes to laminin

This study investigated the possible relationship between the invasiveness of group A Streptococcus (GAS) strains and their abilities to adhere to laminin and assessed the effects of subinhibitory concentrations of penicillin and erythromycin on the ability of GAS to adhere to laminin. The adherence of noninvasive and highly invasive isolates of GAS to laminin was significantly higher than the adherence displayed by isolates of low invasiveness. Antibiotic treatment caused significant reductions in adherence to laminin in all three groups of strains. Penicillin was more successful in reducing the adherence abilities of the tested GAS strains than erythromycin.

Interferon-γ inhibits group B Streptococcus survival within human endothelial cells

Endothelial dysfunction is a major component of the pathophysiology of septicaemic group B Streptococcus (GBS) infections. Although cytokines have been shown to activate human umbilical vein endothelial cells (HUVECs), the capacity of interferon (IFN)-γ to enhance the microbicidal activity of HUVECs against GBS has not been studied. We report that the viability of intracellular bacteria was reduced in HUVECs activated by IFN-γ. Enhanced fusion of lysosomes with bacteria-containing vacuoles was observed by acid phosphatase and the colocalisation of Rab-5, Rab-7 and lysosomal-associated membrane protein-1 with GBS in IFN-γ-activated HUVECs. IFN-γ resulted in an enhancement of the phagosome maturation process in HUVECs, improving the capacity to control the intracellular survival of GBS.

Newly-synthesized chalcones-inhibition of adherence and biofilm formation of methicillin-resistant Staphylococcus aureus

Biofilm formation and adherence of bacteria to host tissue are one of the most important virulence factors of methicillin-resistant strains of Staphylococcus aureus (MRSA). The number of resistant strains is seriously increasing during the past years and bacteria have become resistant, not only to methicillin, but also to other commonly used antistaphylococcal antibiotics. There is a great need for discovering a novel antimicrobial agent for the treatment of staphylococcal infections. One of the most promising groups of compounds appears to be chalcones. In present study we evaluated the in vitro effect of three newly synthesized chalcones: 1,3-Bis-(2-hydroxy-phenyl)-propenone, 3-(3Hydroxy-phenyl)-1-(2-hydroxy-phenyl)-propenone and 3-(4-Hydroxy-phenyl)-1-(2-hydroxyphenyl)-propenone on glycocalyx production, biofilm formation and adherence to human fibronectin of clinical isolates and laboratory control strain of MRSA (ATCC 43300). Subinhibitory concentrations of the tested compounds reduced the production of glycocalyx, biofilm formation and adherence to human fibronectin of all MRSA strains. Inhibition of biofilm formation was dose dependent and the most effective was 1,3-Bis-(2-hydroxy-phenyl)-propenone. In our study we demonstrated that three newly-synthesized chalcones exhibited significant effect on adherence and biofilm formation of MRSA strains. Chalcones may be considered as promising new antimicrobial agents that can be used for prevention of staphylococcal infections or as adjunct to antibiotics in conventional therapy.

[Study in vitro study of the efficacy of some antibiotic in preventing the adherence of some aerobic bacteria isolated from tonsil's core]

It was proven that biofilm formation had an important role in relapse infections and bacterial resistance against antibiotics in recurrent bacterial infections. Bacterial biofilm formation in crypts of tonsil's Core is considered as a major factor in the chronicity and treatment failure in patients with recurrent tonsillitis. According to that, we aimed in our study to evaluate the efficacy of some antibiotics recommended in recurrent tonsillitis in preventing the biofilm formation by some bacteria isolated from tonsil's core. We had worked on five aerobic species, isolated from tonsil's core of children suffering from recurrent tonsillitis and undergoing tonsillectomy, three were Gram Positive Cocci [Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus pyogenes], and two were Gram Negative bacilli [Klebsiella pneumonia, Escherichia coli]. We evaluated efficacy of five different antibiotics, Amoxicillin clavulanic acid, Azithromycin, Lincomycin, Cefuroxime, and Ceftriaxone. Bacterial identification and Disk diffusion test were carried out through standard procedures. Evaluation of the anti adherence efficacy of the antibiotics were carried out by using "The microtiter plate assay". Our study shows that Azithromyin has the best efficacy on the Gram Positive Cocci adherence, followed by Cephalosporin, and Augmentin, while Lincomycin increased the adherence's rate. Ceftriaxone has the best efficacy on the Gram Negative Bacilli adherence, while Lincomycin and Cefuroxime increased the adherence's rate, and Augmentin and Azithromycin have no effect on the adherence

Heparan sulphate, its derivatives and analogues share structural characteristics that can be exploited, particularly in inhibiting microbial attachment

Heparan sulphate (HS) and the related polysaccharide, heparin, exhibit conformational and charge arrangement properties, which provide a degree of redundancy allowing several seemingly distinct sequences to exhibit the same activity. This can also be mimicked by other sulphated polysaccharides, both in overall effect and in the details of interactions and structural consequences of interactions with proteins. Together, these provide a source of active compounds suitable for further development as potential drugs. These polysaccharides also possess considerable size, which bestows upon them an additional useful property: the capability of disrupting processes comprising many individual interactions, such as those characterising the attachment of microbial pathogens to host cells. The range of involvement of HS in microbial attachment is reviewed and examples, which include viral, bacterial and parasitic infections and which, in many cases, are now being investigated as potential targets for intervention, are identified.

In vitro oxidant effects of D-glucosamine reduce adhesión and biofilm formation of Staphylococcus epidermidis / Los efectos oxidantes de la D-glucosamina in vitro reducen la adhesión y formación de biofilms de Staphylococcus epidermidis

Staphylococcus epidermidis is a common pathogen in medical device-associated infections. Its major pathogenic factor is the ability to form adherent biofilms. In this work, three S. epidermidis strains isolated from infected catheters were chosen with the objective of investigating the effect of D-glucosamine (D-Glu) on reactive oxygen species (ROS) production, adhesion and biofilm formation. The chemiluminescence and nitroblue tetrazolium reduction assays were used to determine ROS production by planktonic S. epidermidis and the microtiter plate assay to quantify in vitro biofilm formation. D-Glu generated a dose-dependent increase in ROS in planktonic cells with maximum stimuli at a concentration of 0.05 mM, and reduced adhesion and biofilm formation. On the other hand, glucose showed an antioxidative stress action and promoted biofilm adhesion and growth. This study suggests a potential application of D-Glu against infections associated with indwelling medical devices, since the oxidative stress caused by this hexosamine in planktonic S. epidermidis contributed to reducing biofilm formation.

Staphylococcus epidermidis es un patógeno común en infecciones asociadas a dispositivos médicos. Su factor de patogenicidad más importante es la capacidad para formar biofilms. Se trabajó con tres cepas de S. epidermidis aisladas de catéteres, con las que se efectuaron ensayos de quimioluminiscencia y de reducción de azul de nitrotetrazolio, para determinar la producción de especies reactivas del oxígeno (ERO) en S. epidermidis planctónico, y ensayos dirigidos a cuantificar la formación de biofilm in vitro, empleando placas multipocillos. La D-glucosamina generó un aumento dependiente de la dosis en la producción de ERO en las células planctónicas, con un estímulo máximo a una concentración de 0,05 mM. Este aumento condμlo a la reducción de la adhesión y de la formación de biofilm. La adición de glucosa, en cambio, mostró un efecto anti estrés oxidativo y promovió la adhesión y el crecimiento de biofilm. Este estudio sugiere una posible aplicación de la D-glucosamina contra las infecciones asociadas a dispositivos médicos, ya que el estrés oxidativo provocado por esta hexosamina contribuyó a una menor formación de biofilm.

Cranberries and lower urinary tract infection prevention

Lower urinary tract infections are very common diseases. Recurrent urinary tract infections remain challenging to treat because the main treatment option is long-term antibiotic prophylaxis; however, this poses a risk for the emergence of bacterial resistance. Some options to avoid this risk are available, including the use of cranberry products. This article reviews the key methods in using cranberries as a preventive measure for lower urinary tract infections, including in vitro studies and clinical trials.