Prevalence of Candida spp. associated with bacteria species on complete dentures

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Er : YAG laser: Antimicrobial effects in the root canals of dogs' teeth with pulp necrosis and chronic periapical lesions

Objective: Our goal in this study was to evaluate the antimicrobial effect of Er:YAG laser applied after biomechanical preparation of the root canals of dog's teeth with apical periodontitis. Background Data: Various in vitro studies have reported effective bacterial reduction in infected root canals using Er:YAG laser. However, there is no in vivo research to support these results. Methods: Forty root canals of dogs' premolar teeth with pulp necrosis and chronic periapical lesions were used. An initial microbiological sample was taken, and after biomechanical preparation was carried out, a second microbiological sample was taken. The teeth were divided into two groups: Group I-biomechanical preparation was taken of root canals without Er:YAG laser application; Group II-biomechanical preparation was taken of root canals with Er:YAG laser application using 140-mj input, 63-mJ output/15 Hz. After coronal sealing, the root canals were left empty for 7 days at which time a third microbiological sample was taken. The collected material was removed from the root canal with a #40 K file and placed in transport media. It was serially diluted and seeded on culture dishes selective for anaerobes, aerobes, and total streptococci. Colony-forming units per milliliter (CFU/mL) were counted. Results: Groups I and II showed an increase of CFU/mL for all microorganisms 7 days after treatment, being statistically significant for anaerobes in Group I and for anaerobes and total streptococci in Group II. When comparing CFU/mL of Groups I and II, there was a statistically significant increase after 7 d for total streptococci in Group II. Conclusion: Er:YAG laser applied after biomechanical preparation did not reduce microorganisms in the root canal system.

Effectiveness of 2% peracetic acid for the disinfection of gutta-percha cones

The aim of this study was to evaluate the effectiveness of 2% peracetic acid for the disinfection of gutta-percha cones contaminated in vitro with Escherichia coli, Staphylococcus aureus, Streptococcus mutans, Candida albicans and Bacillus subtilus (in spore form). Two hundred and twenty-five gutta-percha cones were contaminated with standardized suspensions of each microorganism and incubated at 37 degrees C for 24 h. The cones were divided into 10 experimental groups (n = 15), according to the microorganism tested and disinfection testing times. The disinfection procedure consisted of immersing each cone in a plastic tube containing the substance. The specimens remained in contact with the substance for 1 or 2.5 minutes. Afterwards, each cone was transferred to a 10% sodium thiosulphate solution (Na2S2O3) to neutralize the disinfectant. Microbial biofilms adhering to the cones were dispersed by agitation. Aliquots of 0.1 ml of the suspensions obtained were plated on Sabouraud dextrose agar, or brain and heart infusion agar, and incubated at 37 degrees C for 24 h. The results were expressed in colony forming units (CFU/ml) and the data were submitted to the Wilcoxon Signed Rank Test (level of significance at 0.05). A significant reduction was observed, after 1 minute of exposure, in the test solution for C. albicans (p = 0.0190), S. aureus (p = 0.0001), S. mutans (p = 0.0001), B. subtilis (p = 0.0001), and E. coli (p = 0.0001). After 2.5 minutes of exposure, 100% of the microbial inocula were eliminated. It was concluded that the 2% peracetic acid solution was effective against the biofilms of the tested microorganisms on gutta-percha cones at 1 minute of exposure.

Surface Roughness and Bacterial Adherence to Resin Composites and Ceramics

Purpose: The aim of this study was to evaluate the surface roughness and the in vitro adherence of Streptococcus mutans to indirect aesthetic restorative materials that are uncoated with saliva.Materials and Methods: Four groups of restorative materials were evaluated according to material type: (1) microparticulate feldspathic ceramic; (2) leucite-reinforced feldspathic ceramic; (3) microhybrid resin composite and (4) microfilled resin composite. Twenty standardised samples of each material were produced. Roughness analysis (Ra, n = 10) was performed using a roughness analyser. Adhesion tests (n = 10) were carried out in 24-well plates; colony-forming units (CFU/mL) were evaluated. The mean values of roughness (mu m) and adherence (CFU/mL) for each group were subjected to an analysis of variance and a Tukey test.Results: The leucite-reinforced feldspathic ceramic was rougher and presented higher bacterial adherence than the microparticulate feldspathic ceramic. The resin composites were similar with regard to surface roughness and bacterial adherence.Conclusions: The microhybrid and microfilled resin composites were similar and the leucite-reinforced feldspathic ceramic was rougher and presented higher bacterial adherence than the microparticulate feldspathic ceramic.

Microbiological or Chemical Models of Enamel Secondary Caries Compared by Polarized-Light Microscopy and Energy Dispersive X-ray Spectroscopy

Different secondary caries models may present different results. The purpose of this study was to compare different in vitro secondary caries models, evaluating the obtained results by polarized-light microscopy (PLM), scanning electron microscopy (SEM), and energy dispersive X-ray spectroscopy (EDS). Standardized human enamel specimens (n = 12) restored with different materials (Z250 conventional composite resin-CRZ, Freedom polyacid-modified composite resin-CRF, Vitremer resin-modified glass-ionomer-GIV, and Fuji IX conventional glass-ionomer cement-GIF) were submitted to microbiological (MM) or chemical caries models (CM). The control group was not submitted to any caries model. For MM, specimens were immersed firstly in sucrose broth inoculated with Streptococcus mutans ATCC 35688, incubated at 37 degrees C/5% CO(2) for 14 days and then in remineralizing solution for 14 days. For CM, specimens were submitted to chemical pH-cycling. Specimens were ground, submitted to PLM and then were dehydrated, gold-sputtered and submitted to SEM and EDS. Results were statistically analyzed by Kruskall-Wallis and Student-Newman-Keuls tests (alpha = 0.05). No differences between in vitro caries models were found. Morphological differences in enamel demineralization were found between composite resin and polyacid-modified composite resin (CRZ and CRF) and between the resin-modified glass-ionomer and the glass-ionomer cement (GIF and GIV). GIF showed higher calcium concentration and less demineralization, differing from the other materials. In conclusion, the glass-ionomer cement showed less caries formation under both in vitro caries models evaluated. (C) 2009 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater 90B: 635-640, 2009

Morphology and bacterial colonisation of tooth/ceramic restoration interface after different cement excess removal techniques

Objectives: To evaluate the influence of different protocols for resin cement removal during cementation on biofilm formation.Methods: Twenty-eight ceramic blocks, which were injected under pressure, were placed over enamel blocks obtained from freshly extracted bovine incisors. The ceramic blocks were cemented to the enamel blocks using a dual-cured resin cement and the excess resin was removed according to the experimental group: TS: Teflon spatula; BR: brush; BR+: brush and polishing; SB+: scalpel blade and polishing. After autoclaving, the samples were colonised by incubation in a sucrose broth suspension standardised with Streptococcus mutans in microaerophilic stove. Specimens were quantitatively analysed for bacterial adherence at the adhesive interface using confocal laser scanning microscopy and counting the colony forming units, and qualitatively analysed using SEM. The roughness (Ra/Rz/RSm) was also analysed. Data were analysed by 1-way ANOVA and Tukey's test (5%).Results: The roughness values ranged from 0.96 to 1.69 mu m for Ra (p > 0.05), from 11.59 to 22.80 mu m for Rz (p = 0.02 < 0.05) and from 293.2 to 534.3 mu m for RSm (p = 0.00). Bacterial adhesion varied between 1,974,000 and 2,814,000 CFU/ml (p = 0.00). Biofilm mean thickness ranged from 0.477 and 0.556 mu m (p > 0.05), whilst the biovolume values were between 0.388 and 0.547 mu m(3)/mu m(2) (p = 0.04). Lower values for roughness, bacterial adhesion, biofilm thickness and biovolume were found with BR, whilst TS presented the highest values for most of the parameters. SEM images confirmed the quantitative values.Conclusions: The restoration margin morphology and interface roughness affects bacterial accumulation. The brush technique promoted less bacterial colonisation at the adhesive interface than did the other removal methods.Clinical significance: The brush technique seems to be a good option for removing the excess resin cement after adhesive cementation in clinical practice, as indicated by its better results with lower bacterial colonisation. (C) 2012 Elsevier Ltd. All rights reserved.

In vitro study of the Nd : YAG laser effect on human dental enamel: Optical and scanning electron microscope analysis

Objective: the Nd:YAG laser irradiation of dental enamel was evaluated in enamel demineralization experiments in a Streptococcus mutans culture media. Summary Background Data: Previous studies had shown that a continuous wave Nd:YAG laser at an energy of approximately 67 mJ may induce an increased acid resistance in human dental enamel when exposed to severe demineralization conditions. Methods: Enamel windows of 3 x 4 cm in the buccal surface were irradiated with a continuous wave Nd:YAG laser at a wavelength of 1,064 Ecm using energy densities of from 83.75 to 187.50 J/cm(2), Enamel windows of 3 x 4 cm on the lingual surface served as control (without the laser irradiation). The enamel windows were then exposed to a Streptococcus mutans culture media at a temperature of 37 degrees C for 15 and 21 days. The laser effects and demineralization were examined both by optical microscopy and scanning electron microscopy (SEM), Results: A comparison between the lased and the unlased windows of enamel showed fusion and recrystalization of the enamel and increased acid-resistance in all groups irradiated with the Nd:YAG laser, on the other hand, the 3 x 4 delimited enamel surfaces from the control group (not irradiated with the Nd:YAG laser) showed 100% deminerization, Conclusions: These findings are consistent with the finding that laser irradiation of dental results in significant reduction of the effective solubility of enamel mineral.

Proposal of Using Ozonated Water to Control Biofilm Formation on Mouth-Related Devices

The purpose of this study was to evaluate the antimicrobial effectiveness and dental applications of ozonated water generated by portable equipment (0.667 mg/L). Total elimination of C. albicans, E. coli and S. mutans planktonic cells was observed after 5 min. Reduction in the number of viable cells of biofilms formed on acrylic resin was observed for C. albicans, S. mutans and E. coli. The same effect was observed on biofilms of E. coli, S. aureus, S. mutans and C. albicans formed on stainless steel. Ozonated water was effective for the disinfection of experimentally contaminated toothbrushes.

Salivary parameters and teeth erosions in patients with gastroesophageal reflux disease

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Microbiological quality of recreational waters in Araraquara, SP, Brazil

The microbiological flora of 108 water samples was explored to evaluate the role of recreational waters as a possible source of human diseases in Araraquara, S.P., Brazil. These waters included six swimming pools and three lakes with beaches. The number of total and fecal coliforms, Escherichia coli, fecal streptococci, Pseudomonas aeruginosa, Staphylococcus aureus, Candida albicans, and heterotrophic organisms was determined. As was the occurrence of Salmonella, Shigella, Yersinia, enteropathogenic E. coli (EPEC), enteroinvasive E. coli (EIEC), Mycobacteria, yeasts in general and dermatophytes. Shigella, Yersinia, EIEC and dermatophytes were not isolated. Other organisms or groups of microorganisms were found in variable proportions. From this study it is concluded that recreational waters used by the population of Araraquara, may be contaminated with potentially pathogenic microorganism and this may serve as a source of human diseases.

Preparation and antimicrobial activity of gelatin microparticles containing propolis against oral pathogens

Gelatin microparticles containing propolis ethanolic extractive solution were prepared by spray-drying technique. Particle,, with regular morphology, mean diameter ranging of 2.27 mu m to 2.48 mu m, and good entrapment efficiency for propolis were obtained. The in vitro antimicrobial activity of microparticles was evaluated against microorganisms of oral importance (Enterococcus faecalis, Streptococcus salivarius, Streptococcus sanguinis, Streptococcus mitis, Streptococcus mutans, Streptococcus sobrinus, Candida albicans, and Lactobacillus casei). The utilized techniques were diffusion in agar and determination of minimum inhibitory concentration. The choice of the method to evaluate the antimicrobial activity of microparticles showed be very important. The microparticles displayed activity against all tested strains of similar way to the propolis, showing greater activity against the strains of E. salivarius, S. sanguinis, S. mitis, and C albicans.

Bacteriocin-like substances production by Streptococcus-salivarius against oral bacteria

Samples of tongue and bucal mucosa surfaces were obtained from six healthy subjects with the purpose of isolating S. salivarius. It was verified that 47 out of 48 S. salivarius strains produced bacteriocin-like substances against at least one of the indicator species: Actinomyces viscosus, Rothia dentocariosa, Streptococcus pyogenes, Staphylococcus aureus, Streptococcus mutans and Streptococcus sanguis. The method employed to test for bacteriocin production was that of deferred antagonism. The results showed that there was a high antagonism against R. dentocariosa, S. pyogenes and A. viscosus; extremely low against S. mutans and S. sanguis and no inhibition for S. aureus.

Isolation of a new L-amino acid oxidase from Crotalus durissus cascavella venom

A novel L-amino acid oxidase (LAO) (Casca LAO) from Crotalus durissus cascavella venom was purified to a high degree of molecular homogeneity using a combination of molecular exclusion and ion-exchange chromatography system. The purified monomer of LAO presented a molecular mass of 68 kDa and pI estimated in 5.43, which were determined by two-dimensional electrophoresis. The 71st N-terminal amino acid sequence of the LAO from Crotalus durissus cascavella presented a high amino acid sequence similarities with other LAOs from Colloselasma rhosostoma, Crotalus adamanteus, Agkistrodon h. blomhoffi, Agkistrodon h. halys and Trimeresurus stejnegeri. LAO displayed a Michaelis-Menten behavior with a kilometer of 46.7 mu M and an optimum pH for enzymatic activity of 6.5. Casca LAO induced a dose-dependent platelet aggregation, which was abolished by catalase and inhibited by indomethacin and aspirin. These results suggest that the production of H2O2 is involved in subsequent activation of inflammatory enzymes, such as thromboxane. Casca LAO also inhibited the bacterial Growth of Gram-negative (Xanthomonas axonopodis pv passiflorae) and Gram-positive (S. mutans) strains. Electron microscopy assessments of both bacterial strains suggest that the hydrogen peroxide produced by LAO induce bacterial membrane rupture and consequently loss of cytoplasmatic content. This LAO exhibited a high antileishmanic activity against the promastigote of Leishmania amazonensis in vitro, its activity was dependent on the production of hydrogen peroxide, and the 50% inhibitory concentration was estimated in 2.39 mu g/ml. (C) 2005 Elsevier Ltd. All rights reserved.

Atividade antimicrobiana de extratos brutos de Petiveria alliacea L.

"Antimicrobial activity of crude extratcs of Petiveria alliacea L.". Petiveria alliacea L. (Phytolaccacea) is an herbaceous plant of great importance in traditional medicine. This species have been widely used in several applications such as antirheumatic, anticarcinogenic, anti-flu, antitussive, analgesic, insecticidal, acaricidal, as well as bactericide and fungicide. Currently, the pathogenic microorganisms are acquiring resistence against the traditional antibiotics, and the search for new herbal antimicrobial agents has been intensified. The objective of this study was to evaluate the antifungal and antibacterial activity of several leaf crude extracts of P. alliacea against several strains of bacterias and yeasts namely Bacilus subtilis, Pseudomonas aeruginosa, Escherichia coli, Streptococcus mutans, Staphylococcus aureus, Staphylococcus epidermidis, Enterococcus faecalis, Candida parapsilosis, Candida kefyr and Candida albicans, using microdilution method. Promising results were observed for the 70% v/v ethanolic extract which presented minimum inhibitory concentration (MIC) from 250 to 760 mu g/mL for yeast. For the bacteria strains tested the MIC ranged between 240 to 3960 mu g/mL, depending of the extractive solution tested.

Antimicrobial photodynamic action on dentin using a light-emitting diode light source

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)