A laboratory investigation on the effect of biguanide- and pyridine-derived antiseptics on the adhesion of resin composites to dentin.

Optimal bonding of adhesive restorations to dentin is crucial to prevent microleakage and enhance the survival of root-filled teeth. The aim of this study was to investigate the effect of chlorhexidine (CHX), alexidine (ALX) and octenidine dihydrochloride (OCT) on the bond strength of resin composites to coronal dentin. Human coronal dentin specimens were treated with 2% CHX, 0.1% ALX, 0.1% OCT or saline then restored with traditional or bulk-fill resin composites. The adhesion strength between the resin and dentin was measured using the microtensile bond strength and failure mode was determined using a stereomicroscope. Treatment with ALX and OCT resulted in significantly greater µTBS compared with CHX and saline, irrespective of the resin composite used. Alexidine treatment predominantly resulted in mixed failure, while adhesive failures were frequently observed in CHX and saline-treated dentin. In conclusion, final irrigation with ALX or OCT improved the bonding of resin composites to dentin.

The presence of Enterococcus faecalis in saliva as a risk factor for endodontic infection.

Aim: The aim of the present study was to investigate and correlate the prevalence of Enterococcus faecalis in saliva and in root canals with different pulpal and periapical conditions. Methodology: Sixty-seven patients were divided into five groups based on pulpal and periapical tissue status: healthy vital teeth (HVT, n=7), healthy treated teeth without lesion (HTT, n=9), irreversible pulpitis (IP, n=13), necrosis (N, n=18), and post-treatment apical periodontitis (PTAP, n=20). Saliva, rubber dam, sterility control and pre-treatment root canal samples were collected and microbiologically processed by culture method. The phylogenetic relationship of E. faecalis isolates collected from root canals and saliva were investigated by whole genome sequencing. Fisher's exact test was used to correlate the presence of E. faecalis in root canals or saliva with clinical and/or radiographic findings. Linear/logistic regression analyses were performed to establish the relationship between the presence of E. faecalis in root canals, saliva, and the status of periapical tissues. Results: E. faecalis was found in 18 root canal and saliva samples. E. faecalis root canal isolates were recovered with the highest frequency from post-treatment apical periodontitis. The occurrence of E. faecalis in saliva was strongly associated with its detection in the root canals (P < 0.001). The pretreatment presence of E. faecalis in root canals was associated with significantly higher odds of having periapical lesions (OR=11.03; 95% CI, 1.27-95.70; p < 0.05). Saliva and root canal isolates from the same patient were highly correlated at the phylogenetic level (Jaccard index >0.95). Conclusion: This pilot study confirms the role of E. faecalis in developing peri-radicular lesions in secondary endodontic infections and suggests that saliva could be the main source of infection. Further studies are needed to investigate the exact origin of this bacteria and its true role in the pathogenesis of secondary/persistent endodontic infections.

Trans-cinnamaldehyde-Biosurfactant Complex as a Potent Agent against Enterococcus faecalis Biofilms.

Enterococcus faecalis is an opportunistic microbial pathogen frequently associated with diverse infections, including those of the skin and teeth, as well as those from surgical wounds. It forms robust biofilms that are highly tolerant to most antimicrobials and first-line antibiotics. Therefore, investigating alternative strategies to eradicate its biofilms is a critical need. We recently demonstrated that trans-cinnamaldehyde (TC) potently kills E. faecalis biofilm cells and prevents biofilm recovery, and yet, the extreme hydrophobicity of TC hampers clinical translation. Here, we report that a complex of TC with an FDA-approved biosurfactant (acidic sophorolipid/ASL) significantly reduces the bacterial viability and biomass of E. faecalis biofilms, compared to TC alone. A confocal laser-scanning microscopic analysis demonstrated that the TC-ASL treatment significantly decreased the biofilm thickness and volume. In conclusion, our study highlights the anti-biofilm potential of the newly developed TC-ASL.

Transition metals and Enterococcus faecalis: Homeostasis, virulence and perspectives.

Enterococcus faecalis, a Gram-positive bacterium, is known to be a key player in several chronic infections as well as nosocomial, heart valve, urinary tract, surgical wound, and dental root canal infections. The capability to sense different transition metal levels and tune its response accordingly endows it with the potential to thrive and cause infections in several host niches. Over the past decade, our knowledge of how transition metals play a critical role in maintaining homeostasis of E. faecalis has improved significantly. The aim of this review is to elucidate the roles of metals such as iron, manganese, zinc, and copper in the physiology, metabolism, and pathogenicity of E. faecalis. These essential micronutrients contribute to energy production, redox stress response, expression of virulence determinants, and cooperation in polymicrobial communities. The review also highlights metal homeostasis systems in E. faecalis, which respond to fluctuations in extracellular metal levels, and regulate the intracellular metal content. Regulation of intracellular metallome secures the tolerance of E. faecalis to oxidative stress and host-mediated metal sequestration strategies. Therapeutic interventions which deprive E. faecalis of its essential metal requirements or disrupt its homeostatic control have been proposed to combat E. faecalis infections.

Antibiofilm activity of phytochemicals against Enterococcus faecalis: A literature review.

Enterococcus faecalis is a leading causative pathogen of recalcitrant infections affecting heart valves, urinary tract, surgical wounds and dental root canals. Its robust biofilm formation, production of virulence factors and antibiotic resistance contribute significantly to its pathogenicity in persistent infections. The decreased effectiveness of most of antibiotics in preventing and/or eradicating E. faecalis biofilms mandates the discovery of alternative novel antibiofilm agents. Phytochemicals are potential sources of antibiofilm agents due to their antivirulence activity, diversity of chemical structure and multiple mechanisms of action. In this review, we describe the phenotypic and genetic attributes that contribute to antimicrobial tolerance of E. faecalis biofilms. We illuminate the benefits of implementing the phytochemicals to tackle microbial pathogens. Finally, we report the antibiofilm activity of phytochemicals against E. faecalis, and explain their mechanisms of action. These compounds belong to different chemical classes such as terpenes, phenylpropenes, flavonoids, curcuminoids and alkaloids. They demonstrate the ability to inhibit the formation of and/or eradicate E. faecalis biofilms. However, the exact mechanisms of action of most of these compounds are not fully understood. Therefore, the future studies should elucidate the underlying mechanisms in detail.

A micro-computed tomographic analysis of obturation quality and retreatability of an epoxy resin-based sealer.

BACKGROUND: To evaluate the voids percentage and the retreatability by different supplementary irrigation techniques of a matched-taper single cone and epoxy resin-based sealer. METHODS: A total of 72 single-rooted premolars were prepared and filled with gutta-percha and MM-Seal. The percentage of voids in the whole root canal and each anatomical part were evaluated by Micro-CT scans. After root filling removal, the teeth were randomly allocated (N.=24) to group A, syringe irrigation; group B, Tornado Brush, and group C, ultrasonically activated irrigation and re-scanned to establish the volume of remnant root filling material. The data relative to voids and retreatment techniques were established by the Kruskal-Wallis test. Differences in retreatment time were examined using one-way ANOVA and Tukey's Test (P<0.05). RESULTS: No significant difference was found in the voids percentage between the different root-thirds and between the supplementary irrigation groups in the MM-Seal remnants for the whole root canal and each section. In all groups, the coronal region exhibited more remaining material than other sections. CONCLUSIONS: MM-Seal showed low voids percentage and its removal was not significantly improved by supplementary irrigation techniques.

Fsr quorum sensing system modulates the temporal development of Enterococcus faecalis biofilm matrix.

Quorum sensing (QS) is a cell-to-cell communication process that regulates major pathogenic attributes in bacteria including biofilm formation, secretion of virulence factors, and antimicrobial resistance. The two-component Fsr-QS system of the nosocomial pathogen Enterococcus faecalis controls the production of extracellular gelatinase that contributes to biofilm development by enhancing the release of nucleic acids into the biofilm matrix. However, the contribution of this system to the deposition of other biofilm matrix components such as polysaccharides and proteins remains unknown. Using wild type and mutant strains, we discovered that biofilm formation was attenuated by inactivation of the Fsr system or its downstream gelatinase production. Inactivation of the Fsr system caused a modest, yet significant reduction in biofilm metabolic activity without affecting cell counts. Inactivation of the QS-signal sensor FsrC and response regulator FsrA resulted in decreased extracellular polysaccharides and proteins in biofilms in a temporal manner. Irrespective of biofilm age, eDNA levels were reduced in the gelatinase mutant strain. Our results collectively suggest that the Fsr system contributes to the temporal deposition of polysaccharides and proteins into the extracellular polymeric matrix (EPS) of E. faecalis biofilm, without affecting bacterial viability. This understanding of the role of the Fsr-QS system in biofilm development may reveal a novel target to develop effective antibiofilm agents to tackle E. faecalis-mediated infections such as in dental root canals, heart valves, and surgical sites.

Trans-Cinnamaldehyde Attenuates Enterococcus faecalis Virulence and Inhibits Biofilm Formation.

Enterococcus faecalis as an important nosocomial pathogen is critically implicated in the pathogenesis of endocarditis, urinary tract, and persistent root canal infections. Its major virulence attributes (biofilm formation, production of proteases, and hemolytic toxins) enable it to cause extensive host tissue damage. With the alarming increase in enterococcal resistance to antibiotics, novel therapeutics are required to inhibit E. faecalis biofilm formation and virulence. Trans-cinnamaldehyde (TC), the main phytochemical in cinnamon essential oils, has demonstrated promising activity against a wide range of pathogens. Here, we comprehensively investigated the effect of TC on planktonic growth, biofilm formation, proteolytic and hemolytic activities, as well as gene regulation in E. faecalis. Our findings revealed that sub-inhibitory concentrations of TC reduced biofilm formation, biofilm exopolysaccharides, as well as its proteolytic and hemolytic activities. Mechanistic studies revealed significant downregulation of the quorum sensing fsr locus and downstream gelE, which are major virulence regulators in E. faecalis. Taken together, our study highlights the potential of TC to inhibit E. faecalis biofilm formation and its virulence.

The Influence of Surface Roughening and Polishing on Microbial Biofilm Development on Different Ceramic Materials.

PURPOSE: To test the hypothesis that surface roughening and polishing of ceramics have no effect on their surface roughness and biofilm adhesion. MATERIALS AND METHODS: Feldspathic ceramic Vitablocks™ TriLuxe forte (VTF), lithium disilicate glass IPS e.max Press™ (IPS) and zirconia reinforced lithium silicate Vita Suprinity™ (VS) ceramic blocks (n = 27 per group) were prepared from sintered CAD blocks using a water-cooled saw. They were further subdivided into 3 subgroups according to the surface treatment protocols (n = 9): as prepared, roughened and polished. The surface roughness of the ceramic blocks was measured using an electro-mechanical profilometer. The ceramic sections were inoculated with Streptococcus mutans and incubated for 48 hours to form a biofilm. The ceramic surfaces with the biofilms were analyzed using Confocal Laser Scanning Microscopy to calculate the percentage of live bacteria and substratum coverage by the biofilm, and further visualized using scanning electron microscopy. Statistical analysis was done with SPSS software using two-way ANOVA, followed by post hoc Bonferroni test to identify significant differences between the groups. The level of significance was set at p = 0.05. RESULTS: As prepared VTF showed significantly higher mean surface roughness values than as prepared IPS and VS. The mean percentage of live bacteria and biofilm coverage of the substrate were significantly higher in the roughened ceramic blocks than the as prepared and polished blocks for all three ceramic types (p < 0.05). Polished specimens of VS significantly lower percentage of biofilm coverage than the other groups (p < 0.05). CONCLUSIONS: This study sheds new light that adjustments of ceramic restorations prior to cementation increases the likelihood for formation and adhesion of microbial biofilms on the surface. Polished zirconia reinforced lithium disilicate ceramics demonstrated the lowest bacterial adhesion among the evaluated ceramics.

Trans-cinnamaldehyde potently kills Enterococcus faecalis biofilm cells and prevents biofilm recovery.

Enterococcus faecalis is a biofilm-forming, nosocomial pathogen that is frequently isolated from failed root canal treatments. Contemporary root canal disinfectants are ineffective in eliminating these biofilms and preventing reinfection. As a result, there is a pressing need to identify novel and safe antibiofilm molecules. The effect of short-term (5 and 15 min) and long-term (24 h) treatments of trans-cinnamaldehyde (TC) on the viability of E. faecalis biofilms was compared with currently used root canal disinfectants. Treatment for 15 min with TC reduced biofilm metabolic activity as effective as 1% sodium hypochlorite and 2% chlorhexidine. Treatment with TC for 24 h was significantly more effective than 2% chlorhexidine in reducing the viable cell counts of biofilms. This serendipitous effect of TC was sustained for 10 days under growth-favoring conditions. For the first time, our study highlights the strong antibacterial activity of TC against E. faecalis biofilms, and notably, its ability to prevent biofilm recovery after treatment.

Light Activated Disinfection in Root Canal Treatment-A Focused Review.

Light activated disinfection (LAD) is a strategy for optimizing root canal disinfection by using a highly-selective, targeted killing of bacteria using a combination of photosensitizers and light. Over the past decade, numerous in vitro and clinical studies have been performed to demonstrate the effectiveness of this mode of root canal disinfection. While most studies offer an important understanding of the effectiveness of LAD on monospecies biofilms, few have offered credence to the fact that infections of the root canal system are mediated by polymicrobial biofilms. Hence, it is imperative to understand the effect of LAD on polymicrobial biofilms both in terms of microbial killing and the changes in the biofilm architecture. The aim of this review was to systematically review the literature to evaluate the effect of LAD on dual and multispecies biofilms and demonstrate the antibiofilm effect of LAD. Two databases (PubMed and Scopus) were searched to identify eligible studies using a combination of key words. These studies were reviewed to draw conclusions on the effect of LAD on dual and multi species biofilm and the antibiofilm effect of LAD. It was found that LAD alone may be unable to eradicate dual and multispecies biofilms, but it may enhance the effect of conventional canal debridement strategies. Novel formulations of photosensitizers with nanoparticles showed the potential to inhibit biofilm formation and/or disrupt the biofilm architecture.