Saulo Roni Moraes

Antimicrobial Activity and Flow Rate of Newer and Established Root Canal Sealers

Endodontic sealers that possess both optimum flow ability and antimicrobial properties may theoretically assist in the elimination of microorganisms located in confined areas of the root canal system. The antimicrobial effects and the flow rate of the following sealers were investigated and compared: Kerr Pulp Canal Sealer EWT, Grossmans Sealer, ThermaSeal, Sealer 26, AH Plus, and Sealer Plus. The agar diffusion test was used to assess the antimicrobial activity of the sealers. In the flow assay, the sealers were placed between two glass slabs and a weight of 500 g was placed on the top of the glass. The diameters of the formed discs were recorded. All root canal sealers tested showed some antimicrobial activity against most of the microorganisms. There were no significant differences between the materials tested (p > 0.05). All root canal sealers also flowed under the conditions of this study. Statistical analysis of the results revealed that AH Plus and Kerr Pulp Canal Sealer EWT had flow values significantly superior to the other sealers tested (p > 0.05). Taken together, these findings suggest that these sealers have the potential to help in the microbial control in the root canal system.

Comparison of the effectiveness of bacterial culture, 16S rDNA directed polymerase chain reaction, and checkerboard DNA-dNA hybridization for detection of Fusobacterium nucleatum in endodontic infections.

Fusobacterium nucleatum is a Gram-negative, non-spore-forming, nonmotile, obligatory anaerobic rod that is normally isolated from the oral cavity. Epidemiological studies have shown that this species is one of the most prevalent in primary root canal infections. The purpose of this study was to compare the effectiveness of bacteriological culture, 16S rDNA directed polymerase chain reaction and checkerboard DNA-DNA hybridization for detection of F. nucleatum strains in infected teeth associated with periradicular lesions. Thirteen single-root teeth from adult patients, all having carious lesions, necrotic pulps, and radiographic evidence of periradicular bone loss were included in this study. Combining all methods, the results indicated that F. nucleatum was present in approximately 31% (4 of 13) of the specimens. Incidence of F. nucleatum in root canal infections, as evaluated in this study by polymerase chain reaction, culture, and DNA-DNA hybridization, was 15.4%, 15.4%, and 10.0%, respectively. Our data demonstrated that no method used herein could be considered superior for detecting F. nucleatum directly from clinical samples. However, the small number of samples examined and the low prevalence that was observed should be considered.

Bacteroides fragilis isolates compared by AP-PCR.

Bacteroides fragilis is a component of the normal intestinal flora and an important pathogen in nonintestinal endogenous infections. It has been associated with enteric infections and has already been detected in polluted water. In order to evaluate the genetic diversity of B. fragilis, a total of 31 isolates and two reference strains were examined. This collection included strains from nonintestinal infections [12], intestinal infections [5], intestinal microflora [10], aquatic environments [4], and the reference strains ATCC 25285 and ATCC 23745. DNA fingerprints were detected using two separate PCR reactions with different arbitrary primers. The computer-assisted system Taxotron (Institut Pasteur, Dr P. Grimont) was used to analyze the profiles obtained and dendrograms were generated. By using a distance of 0.65 as the threshold, two clusters (hereafter referred to as genotypes I and II) were defined. Strains of differents origins could be distributed into both genotypes. We were unable to detect any obvious correlation between a given genotype and the specific disease or the source of the corresponding strains.

Surface vesicles : A possible function in commensal relations of Bacteroides fragilis

Surface vesicles (SV) defined by electron microscopy as outer membrane (OM) extrusions were detected in Bacteroides fragilis strains from distinct sources. A partial identity between SV and OM electrophoretic protein profiles, in addition to the microscopic analysis, may suggest the designation of OMSV. Sialidase activity, a virulence determinant, was associated with these sub-cellular structures in all the strains, but in an inverse relation to the vesicle quantity per cell. A commensal strain, previously defined as avirulent in an animal model, presented the lowest vesicle-associated sialidase activity and the greatest SV expression as opposed to what happened with clinical and environmental strains. These results seem to suggest that these surface components have a function in commensal stages of B. fragilis.

Whole-cell and Periplasmic Protein Banding Patterns of Environmental and Human Bacteroides fragilis Strains

In order to investigate the relationship among virulent and avirulent Bacteroides fragilis strains, SDS-PAGE of whole-cell proteins (WP) and periplasmic proteins (PP) were used to establish a protein profile of strains isolated from human infections, fecal flora and environmental water. Despite different sources of the strains, no significant differences were observed as determined by the WP SDS-PAGE analysis. In contrast, the proteins obtained from the bacterial periplasm showed differences in the electrophoretic protein profile. Two distinct PP profile patterns were obtained. Pattern A included 6 out of the 8 virulent strains and pattern B, 6 out of 8 avirulent strains. Interestingly, an environmental strain that was capable of inducing abscesses in mice, had a PP profile highly similar to that of the virulent strains from human infections. These data indicate that PP from B. fragilis may be useful to characterize differences among virulent and avirulent strains. Moreover, strains isolated from environmental water may also be a source of exogenous infections by B. fragilis.

Electrophoretic characterization of exposed outer membrane proteins in environmental and human Bacteroides fragilis strains

Bacteriodes fragilis isolated from aquatic environment, from infectious process and from human feces were compared as to their outer membrane protein electrophoretic profiles after staining with Coomassie blue and reacting with antibodies prepared against whole-cell antigens of a reference strain from a clinical source. A marked homogeneity was found among the strains with these methodologies. The profiles of all strains obtained after radio-iodination of the intact cell showed qualitative similarity when compared with the profiles obtained by the other methods. Thus, these data allow us to suggest the designation of the peptides observed in the autoradiograms as surface-exposed proteins. Differences observed in the autoradiograms in the expression of bands mainly detected at a molecular weight of 28 in the commensal strain 118,310 defined previously as avirulent, in addition to a distinction in the titres of agglutination with the sera tested and lower reactivity in the immunoblotting assays, suggest a relationship of the B. fragilis surface architecture with the virulence potential as well as with the origin of the strain.