Endodontic E. faecalis isolates exhibit high level of resistance to tetracycline, an antibiotic that has use in local treatment of dental infections. This opens up a much-needed debate on the role and efficacy of this antibiotic for oral infections. Furthermore, these isolates were shown to possess genes that could contribute to pathogenicity in the pulp cavity.

Objectives:To determine whether phenotypic and genotypic differences amongst isolates of Enterococcus faecalis relate to geographical and clinical origin. Methods: E. faecalis from primary endodontic infections in Brazilian patients (n=20), oral infections in UK patients (n=10), and non-oral infections in Japanese patients (n=9) were studied. In addition, 20 environmental vancomycin resistant Enterococcus faecalis (VRE) isolates from a UK hospital were analysed. For all isolates, polymerase chain reaction (PCR) was used to detect genes associated with antibiotic resistance and virulence, whilst randomly amplified polymorphic DNA-PCR (RAPD-PCR) was used to produce molecular profiles. Results: Gelatinase gene (gelE) was prevalent amongst isolates (77-100%) and for oral isolates, genes of aggregation substances (agg), immune evasion protein (esp), cytolysin (cylB), tetracycline resistance (tetM; tetL) and erythromycin resistance (ermB) were detected to varying extent. Japanese non-oral isolates had a similar genetic profile to oral isolates, but with higher prevalence of ermB and cylB. All VRE isolates were positive for gelE, esp, agg, vanA, ermB and tetM, 95%were positive for cylB and 17% positive for tetL. All isolates were negative for ermA, asa373 vanB, vanC1 and vanC2/3. RAPD-PCR revealed clustering of VRE isolates.Conclusions: RAPD-PCR analysis revealed extensive genetic variability among the tested isolates. Oral isolates carried antibiotic resistance genes for tetracycline and whilst they possessed genes that could contribute to pathogenicity, these were detected at lower incidence compared with non-oral and VRE isolates. RAPD-PCR proved to be a useful approach to elucidate relatedness of disparate isolates.

ExoU, a Pseudomonas aeruginosa cytotoxin injected via the type III secretion system into host cells, possesses eicosanoid-mediated proinflammatory properties due to its phospholipase A(2) (PLA(2)) activity. This report addressed the question whether ExoU may modulate the expression of adhesion molecules in host cells, therefore contributing to the recruitment of leukocyte into infected tissues. ExoU was shown to down-regulate membrane-bound ICAM-1 (mICAM-1) and up-regulate the release of soluble ICAM-1 (sICAM-1) from P. aeruginosa-infected endothelial cells. The modulation of ICAM-1 depended on the direct effect of the ExoU PLA(2) activity and involved the cyclooxygenase (COX) pathway. No differences in mICAM-1 and sICAM-1 mRNA levels were observed when cultures were infected with the ExoU-producing PA103 strain or the mutant PA103DeltaexoU, suggesting that ExoU may proteolytically cleave mICAM-1, producing sICAM-1 in a COX-dependent pathway.

This study evaluated, in vitro, the cytotoxicity of six root canal sealers after 12, 24 and 72 h of contact time, using an endothelial ECV-304 cell line. The MTT assay was used for analysis of cell viability. Twelve specimens of each sealer were prepared and randomly assigned to 6 groups according to the commercial brands (n=4/time). A control group was also formed, which was not subjected to the contact with sealers. To assess the effects of sealers on endothelial cells, the specimens were placed in culture plate wells and incubated at 37°C with 5% CO 2 and 100% humidity. MTT assays were performed in quadruplicate after 12, 24 and 72 h of contact of the sealer specimens with monolayers. Statistical analysis was performed by two-way ANOVA with Bonferroni post-hoc test at a significance level of 5%. Analysis of absorbance in the experimental groups showed that GuttaFlow presented the lowest cytotoxicity, with a mean absorbance of 0.048, followed by Pulp Canal Sealer (0.038), Sealer 26 (0.038), Endo Densell (0.036) and Pulp Fill (0.035). The control group had a mean absorbance of 0.098. Based on the results, Endofill and GuttaFlow were the most and the least cytotoxic sealers, respectively.

Objective The aim of present study was to assess the cytotoxicity and antimicrobial efficacy of 2% peracetic acid (PAA) compared with 5.25% sodium hypochlorite (NaOCl) and 2% chlorhexidine (CHX). Material and Methods For the cytotoxicity test, 100 µl of the tested solutions were added in 12 wells with ECV 304 endothelial cells in each group: NaOCl, CHX and PAA, in addition to the control group. Each solution was evaluated after 24 hours of contact in four dilutions: 0.2, 0.1, 0.05 and 0.025 through mitochondrial function using MTT colorimetric assay. In the antimicrobial evaluation, 40 dentin blocks 5 mm in length and 0.2 g in weight were incubated with 400 µl of Enterococcus faecalis suspension for 21 days at 37°C. The contaminated samples were divided into three experimental groups within 5 minutes of contact: NaOCl group, CHX group, PAA group, as well as the positive control group. The specimens received treatment and were transferred to a tube with saline for serial dilution of the solution and seeding for isolation and colony forming unit (CFU) count. Statistical Analysis The results obtained were expressed as mean (A570 nm) ± standard deviation (SD) and in a multiple linear regression model and multiple comparisons conducted. Results The antimicrobial evaluation revealed that the NaOCl and CHX groups showed a statistically significant difference compared with the control group (p < 0.001), while the PAA reduced only the CFU growth. It can be concluded that, among the agents tested, PAA expressed greater cell viability, followed by CHX and NaOCl. However, it did not show greater antimicrobial activity in vitro in the mature biofilm of Enterococcus faecalis.