The Multivalent Role of Fibronectin-Binding Proteins A and B (FnBPA and FnBPB) of Staphylococcus aureus in Host Infections

Staphylococcus aureus, one of the most important human pathogens, is the causative agent of several infectious diseases including sepsis, pneumonia, osteomyelitis, endocarditis and soft tissue infections. This pathogenicity is due to a multitude of virulence factors including several cell wall-anchored proteins (CWA). CWA proteins have modular structures with distinct domains binding different ligands. The majority of S. aureus strains express two CWA fibronectin (Fn)-binding adhesins FnBPA and FnBPB (Fn-binding proteins A and B), which are encoded by closely related genes. The N-terminus of FnBPA and FnBPB comprises an A domain which binds ligands such as fibrinogen, elastin and plasminogen. The A domain of FnBPB also interacts with histones and this binding results in the neutralization of the antimicrobial activity of these molecules. The C-terminal moiety of these adhesins comprises a long, intrinsically disordered domain composed of 11/10 fibronectin-binding repeats. These repetitive motifs of FnBPs promote invasion of cells that are not usually phagocytic via a mechanism by which they interact with integrin α5β1 through a Fn mediated-bridge. The FnBPA and FnBPB A domains engage in homophilic cell-cell interactions and promote biofilm formation and enhance platelet aggregation. In this review we update the current understanding of the structure and functional properties of FnBPs and emphasize the role they may have in the staphylococcal infections

Mechanostability of the Fibrinogen Bridge between Staphylococcal Surface Protein ClfA and Endothelial Cell Integrin αVβ3

Binding of the Staphylococcus aureus surface protein clumping factor A (ClfA) to endothelial cell integrin αVβ3 plays a crucial role during sepsis, by causing endothelial cell apoptosis and loss of barrier integrity. ClfA uses the blood plasma protein fibrinogen (Fg) to bind to αVβ3 but how this is achieved at the molecular level is not known. Here we investigate the mechanical strength of the three-component ClfA-Fg-αVβ3 interaction on living bacteria, by means of single-molecule experiments. We find that the ClfA-Fg-αVβ3 ternary complex is extremely stable, being able to sustain forces (∼800 pN) that are much stronger than those of classical bonds between integrins and the Arg-Gly-Asp (RGD) tripeptide sequence (∼100 pN). Adhesion forces between single bacteria and αVβ3 are strongly inhibited by an anti-αVβ3 antibody, the RGD peptide, and the cyclic RGD peptide cilengitide, showing that formation of the complex involves RGD-dependent binding sites and can be efficiently inhibited by αVβ3 blockers. Collectively, our experiments favor a binding mechanism involving the extraordinary elasticity of Fg. In the absence of mechanical stress, RGD572-574 sequences in the Aα chains mediate weak binding to αVβ3, whereas under high mechanical stress exposure of cryptic Aα chain RGD95-97 sequences leads to extremely strong binding to the integrin. Our results identify an unexpected and previously undescribed force-dependent binding mechanism between ClfA and αVβ3 on endothelial cells, which could represent a potential target to fight staphylococcal bloodstream infections

Antibacterial activity of different root canal sealers against Enterococcus faecalis

The aim of the present study was to compare in vitro the antimicrobial activity of different root canal sealers against Enterococcus faecalis, prior and subsequent to setting. Agar diffusion test (ADT) was used for evaluating the antibacterial activity of non-set sealer while the direct contact test (DCT) was used for after setting. ADT: Except for TotalFill BC Sealer all the others sealers tested showed antibacterial activity. BioRoot?RCS, MTA Fillapex and Sealapex Root Canal Sealer showed the lowest antibacterial activity, a significant increase in antibacterial effect for both Pulp Canal Sealer? and AH plus sealers were found. Significantly higher were the mean diameters of the bacterial inhibition zone by both EasySeal or N2 sealers. DCT: AH plus and Sealapex Root Canal Sealer doesn?t show any bactericidal effect after 6 min of contact. After 15 and 60 min of contact a significant increment for AH plus and for Sealapex Root Canal Sealer of the bactericidal effect was found. Significantly much higher was the antibacterial effect of Sealapex Root Canal Sealer compare to that observed for AH plus. BioRootTMRCS, MTA Fillapex, Pulp Canal Sealer? and N2 showed at least means of the number of colonies formed in milliliter after 6 min of contact. Except for N2, a significant increase in bactericidal effect after 15 and 60 min for the other compared sealers (BioRootTMRCS, MTA Fillapex and Pulp Canal Sealer?). For every contact times considered, both TotalFill BC Sealer and EasySeal were bactericidal against E. faecalis and killed all bacteria

Comparative cytotoxicity evaluation of eight root canal sealers

The aim of the present study is to evaluate and compare the cytotoxic effects of eight root canal sealers (BioRoot RCS, TotalFill BC Sealer, MTA Fillapex, Sealapex, AH Plus, EasySeal, Pulp Canal Sealer, N2) on immortalized human gingival fibroblasts over a period of 24, 48 and 72 hours. Immortalized human gingival fibroblast-1 HGF-1 (ATCC CRL-2014) were incubated. Root canal sealers were then placed into sterile, cylindrical Teflon moulds. The extraction was made eluting the sealers in cell culture medium. Cells (1 × 104) were seeded in each well of a 96-well plate and incubated for 24 h at 37°C. Cultures were then exposed to 100 ?L of the extracts medium. The percentage of viable cells in each well was calculated relative to control cells set to 100%. BioRoot RCS and TotalFill BC Sealer extracted for 24h showed no cytotoxic effect, while it was mild by using 48 and 72 h extracts. No cytotoxic effect was measured by using AH Plus medium eluted for 24 h, while it was moderate after 48 h and severe after 72 h. Pulp Canal Sealer, Sealapex and N2 showed moderately cytotoxic activity for all the extraction times. EasySeal and MTA Fillapex remained severely or borderline mildly cytotoxic for all the extraction times. In the present study only BioRoot RCS, TotalFill BC Sealer and AH Plus showed no cytotoxic effects at least in the first 24h. All the other sealers revealed moderately or severely cytotoxic activity during all the extraction times

Protective effect of zinc-hydroxyapatite toothpastes on enamel erosion: an in vitro study

The aim of the present study was to test the impact of different toothpastes with Zinc-Hydroxyapatite (Zn-HAP) on preventing and repairing enamel erosion compared to toothpastes with and without fluoride. The following four toothpastes were tested: two toothpastes with Zn-HAP, one toothpaste with fluoride and one toothpaste without fluoride. An additional control group was used in which enamel specimens were not treated with toothpaste. Repeated erosive challenges were provided by immersing bovine enamel specimens (10 per group) in a soft drink for 2 min (6mL, room temperature) at 0, 8, 24 and 32 h. After each erosive challenge, the toothpastes were applied neat onto the surface of specimens for 3 min without brushing and removed with distilled water. Between treatments the specimens were kept in artificial saliva. Enamel hardness, after the erosive challenge and toothpaste treatment was monitored using surface micro-hardness measurements. As expected, repeated erosive challenge by a soft drink for total of 8 min significantly reduced enamel surface hardness (ANOVA, p < 0.05). No re-hardening of the surface softened enamel was observed in the group treated with fluoride-free toothpaste. Surface hardness of the softened enamel increased when the specimens were treated with the fluoride toothpaste and the two toothpastes with Zn-HAP (p < 0.05). Toothpaste with Zn-HAP resulted in significant enamel remineralisation of erosively challenged enamel, indicating that these toothpastes could provide enamel health benefits relevant to enamel erosion

Evaluation of the antibacterial activity of a new ozonized olive oil against oral and periodontal pathogens

In the present study, the antimicrobial properties of a new ozonized olive oil (O-zone gel) against oral and periodontal pathogens will be evaluated and compared with that of common CHX-based agents. O-zone gel was compared with two agents based on chlorhexidine digluconate (CHX): Corsodyl Dental Gel and Plak Gel. A. actinomycetemcomitans, P. intermedia and S. mutans, were selected and the antibacterial capability of the compounds was tested by using direct contact agar diffusion test (DCT) and minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) evaluations. Differences between specific means were analyzed by a one-way analysis of variance (ANOVA). Group means were compared using a one-way ANOVA and Tukey?s test (P<0.05). O-zone gel reported inhibition zones which correspond to 33% and 43% of that achieved by the CHX agents. No inhibition of bacterial growth (MIC) on the Gram-positive strain by using O-zone gel was found and no antimicrobial effect (MBC) was observed by using O-zone gel on both Gram-negative and -positive strains. The new ozonated oil was a relatively moderate antiseptic. Gram-negative bacteria proved to be more sensitive to ozonized olive oil than Gram-positive ones. The ozonized olive oil demonstrated a lower antibacterial activity if compared to the CHX-based agents tested

Preventive effects of different protective agents on dentin erosion: an in vitro investigation

The purpose of this in vitro study was to evaluate the preventive effects of different protective agents on dentine erosion, measuring mean percentage weight loss. Dissolution of dentine under erosive challenges caused by soft drinks was analyzed: specimens were weighed following each immersion period, with mean percent weight losses calculated. Extracted teeth were sectioned into uniform slabs. Seventy permanent enamel specimens were randomly distributed to seven groups. Initial weights of all dentin specimens were performed. The fluoride pastes Remin Pro, MI Paste Plus, Tooth Mousse, Biorepair, Biorepair Plus and Regenerate were used in this study. A control group was treated just with tap water. The specimens then were immersed in Coca-Cola for a total of 32 min at room temperature. Finally each specimen was dry and weighed. The mass loss was calculated as a percentage of that observed prior the fluoride pastes application. Weight loss data were subjected to Analysis of Variance (One-way ANOVA) followed by Bonferroni’s post hoc tests. Percent weight loss of specimens exposed to early stages in Coca-Cola showed linear progression with time. Specimen’s application of fluoridated varnishes such as Biorepair or Regenerate, prior immersion in Coca-Cola, significantly protect dentin from demineralization. Otherwise, application of Tooth Mousse or Biorepair Plus increased dentin demineralization starting from 24 min of immersion in Coca-Cola. Despite the limitations of this study, the protective pastes that showed the less weight loss due to the acidic challenge are Biorepair and Regenerate

Effect of different protective agents on enamel erosion : an in vitro investigation

The purpose of this in vitrostudy was to compare the effect of different protective agents on enamel erosion by measuring mean percentage weight loss. Extracted teeth were sectioned into uniform slabs and enamel specimens were randomly distributed to different groups. Initial weight of all enamel specimens was registered. The protective agents used in this study were Tooth Mousse, MI Paste Plus, Remin Pro and Remin Pro Forte. A control group was treated just with tap water. All the specimens were immersed in Coca-Cola for a total of 8 min at room temperature, dried and weighed. Enamel dissolution caused by acidic soft drink was analyzed: specimens were weighed after each immersion period and mean percent weight loss was calculated. Weight loss data were subjected to Analysis of Variance (One-way ANOVA) followed by Bonferroni?s post hoc tests. All the groups showed a statistically significant loss of weight (p<0.01) during the testing periods, increased after 8 days (~55%) and 12 days (~70%) of exposure. Specimens treated with protective agents showed significantly lower % of weight loss especially with Remin Pro or Remin Pro Forte. Soft drinks can cause enamel erosion, but protective agents tested may enhance enamel resistance against erosion

Biological and antibacterial properties of a new silver fiber post : in vitro evaluation

The incorporation of nano silver particles (AgNPs) to improve antibacterial properties of dental materials has become increasingly common. The aim of the present study was to compare the antibacterial activity and cytotoxicity effects of different fiber posts: glass fiber post, quartz fiber post, nano fiber post and silver fiber post. The antibacterial activity against S. mutans, S. salivarius and S. sanguis was evaluated by using the agar disc diffusion test (ADT). Four wells of 3x2 mm (one for each material) were made with a punch by removing the agar and filled with the materials to be evaluated. The size of the inhibition zone was calculated. An extract was made eluting the posts in cell culture medium using the surface area-to-volume ratio of approximately 1.25cm²/ml between the surface of the samples and the volume of medium. Cell cultures were then exposed to 100 ?L of the extracts medium. After 24 h, cell viability was determined using the MTT assay. Silver fiber post was the only material showing a fair antibacterial effect against all the three streptococcal strains. The level of cytotoxicity of all the fiber posts tested was higher than 90% and therefore they were considered not cytotoxic. The new silver fiber post reported a fair antibacterial activity. On the other hand all the fiber posts tested (including the post with incorporated AgNPs) proved to be biocompatible, suggesting that their application does not represent a threat to human health

Solubility and pH of bioceramic root canal sealers : a comparative study

Background: This study compared the solubility and the pH of different root canal sealers in vitro . Material and Methods: BioRoot?RCS, TotalFill BC Sealer, MTA Fillapex, SealapexTM, AH Plus, EasySeal, Pulp Canal Sealer? and N2 were tested. Similar specimens were prepared using ring molds with an internal diameter of 20 ± 0,1 mm and a height of 1,5 ± 0,1 mm and digitally weighted to register the mass of each specimen before and after immersion in distilled water. Solubility was determined after 24 hours and statistically analysed using a one-way ANOVA test and post-hoc Tukey test. The pH value was measured by a digital pH meter after 3 and 24 hours from manipulation. Results: BioRoot?RCS and TotalFill BC Sealer showed significantly higher solubility ( P < 0.05). All the remnant root canal sealers fulfilled the requirements of solubility of the International Standard Organization 6876 demonstrating a weight loss of less than 3%. BioRoot?RCS and Totalfill BC Sealer exhibited high alkaline pH over time ( P < 0.05); the alkalinity of the other tested cements was significantly lower . Conclusions: The prolonged alkalinity of bioceramic sealer matched the increase in solubility. This may encourage their biological and antimicrobial effects, but the ongoing solubility may impact their ability to prevent apical leakage