Effect of a Modified Irrigation Protocol on the Cleanliness of Moderately Curved Canals.

OBJECTIVES: This study tested the hypothesis that modifying the sequence of sodium hypochlorite (NaOCl)/ethylene diamine tetra-acetic acid (EDTA) irrigation during root canal shaping would improve apical cleanliness in moderately curved canals. MATERIALS AND METHODS: Forty-five root canals were prepared using Protaper Gold with three irrigation protocols. Standard irrigation (SI) used 0.5 mL 3% NaOCl between each instrument, followed by 5 mL 17% EDTA manually agitated for 30 seconds. Reverse irrigation (RI) used 0.5 mL of 17% EDTA between each instrument, then 5 mL of 3% NaOCl, manually agitated for 30 seconds. Reverse irrigation plus (RI+) was similar to RI, except NaOCl (5 mL), used as a final rinse, was allowed to interact for 3 minutes with dentin before manual agitation (30 seconds).Root canal cleanliness was evaluated under the scanning electron microscope (SEM) (Hulsmann score); the chemical composition of dentin after irrigation was analyzed by energy dispersive X-ray (EDX). STATISTICAL ANALYSIS: Results were compared using Kruskal-Wallis ANOVA by ranks and Wilcoxon matched paired posthoc tests. A Chi-square test assessed whether the best cleanliness would demonstrate a significant association with one irrigation protocol; odds ratio calculation was performed using score "1" versus score "2 or more" (2+) (p < 0.05). RESULTS: In the apical region, cleanliness was better in RI+ than SI and both significantly better than RI. Odd ratios indicate that the cleanliness in RI+ was significantly better than RI and SI groups (p < 0.000 and 0.003, respectively). Independently of the irrigation protocol, EDX analyses showed no chemical alteration of root dentin. CONCLUSIONS: Using 17% EDTA during shaping, followed by 3% NaOCl rinse for 3 minutes, improved apical cleanliness without inducing erosion of dentin.

Root Microbiota in Primary and Secondary Apical Periodontitis.

Apical periodontitis is an inflammatory disease of the dental periradicular tissues triggered by bacteria colonizing necrotic root canals. Primary apical periodontitis results from the microbial colonization of necrotic pulp tissues. Secondary apical periodontitis results from a persistent infection of incorrectly treated root canals. The aim of this study was to characterize the microbiota present in primary and secondary intraradicular infections associated with apical periodontitis using 16S rRNA gene amplicon sequencing. Teeth exhibiting apical periodontitis with or without root canal treatment were extracted after informed consent. From each tooth, the intraradicular content as well as a dentin sample (control) were collected and subjected to DNA extraction. PCR amplicons of the V3-V4 region of the bacterial 16S rRNA gene were pooled and sequenced (2 × 300) on an Illumina MiSeq instrument. The bioinformatics analysis pipeline included quality filtering, merging of forward and reverse reads, clustering of reads into operational taxonomic units (OTUs), removal of putative contaminant OTUs and assigning taxonomy. The most prevalent and abundant OTU in both dentin and root canal samples was assigned to anaerobic bacterium Fusobacterium nucleatum. Multivariate analysis showed clustering of microbiota by sample type (dentin vs. intraradicular content) and, in root canals, by pathology (primary vs. secondary infection). The proportions of Enterococcus faecalis and F. nucleatum were, respectively, higher and lower when comparing secondary to primary infected root canals. Co-occurrence network analysis provided evidence of microbial interactions specific to the infection type. The identification of bacterial taxa differentially abundant in primary and secondary intraradicular infections may provide the basis for targeted therapeutic approaches aimed at reducing the incidence of apical periodontitis.

Enzyme-mediated photoinactivation of Enterococcus faecalis using Rose Bengal-acetate.

Rose Bengal-acetate (RB-Ac) is a pro-photosensitizer claimed to diffuse into target cells, where the acetate groups are hydrolyzed and the photosensitizing properties of Rose Bengal (RB) are restored. Despite promising results on tumor cells, the interaction of RB-Ac with bacteria has never been investigated. This study aimed to assess the interaction of RB-Ac with Enterococcus faecalis and to evaluate its potential use in antimicrobial photodynamic therapy (aPDT). Spectrofluorometry was used to assess the ability of E. faecalis to hydrolyze the RB-Ac compound. Fluorescence microscopy was employed to observe the distribution and to evaluate the cellular uptake of the RB produced. The antibacterial efficiency of RB-Ac-mediated aPDT was assessed by flow cytometry in combination with the LIVE/DEAD® staining. Results showed that RB-Ac was successfully hydrolyzed in the presence of E. faecalis cells. The RB produced appeared to incorporate the membrane of bacteria. Higher concentrations of RB-Ac resulted in higher incorporation of RB. The blue-light irradiation of RB-Ac-treated samples significantly reduced bacterial viability. Less than 0.01% of E. faecalis survived after incubation with 200 µM RB-Ac during 900 min and blue-light activation. The current report indicates that E. faecalis cells can hydrolyze the RB-Ac compound to produce active RB. The use of RB-Ac did not appear to allow cytoplasmic internalization of the RB produced, which rather incorporated the membrane bilayers of E. faecalis. The use of RB-Ac did not provide additional advantages over RB in terms of PS localization. Nonetheless, sufficient RB was produced and incorporated into the membranes of bacteria to elicit effective aPDT.

Rose bengal uptake by E. faecalis and F. nucleatum and light-mediated antibacterial activity measured by flow cytometry.

Antibacterial photodynamic therapy (aPDT) using rose bengal (RB) and blue-light kills bacteria through the production of reactive oxygen derivates. However, the interaction mechanism of RB with bacterial cells remains unclear. This study investigated the uptake efficiency and the antibacterial activity of blue light-activated RB against Enterococcus faecalis and Fusobacterium nucleatum. Spectrophotometry and epifluorescence microscopy were used to evaluate binding of RB to bacteria. The antibacterial activity of RB after various irradiation times was assessed by flow cytometry in combination with cell sorting. Uptake of RB increased in a concentration dependent manner in both strains although E. faecalis displayed higher uptake values. RB appeared to bind specific sites located at the cellular poles of E. faecalis and at regular intervals along F. nucleatum. Blue-light irradiation of samples incubated with RB significantly reduced bacterial viability. After incubation with 10µM RB and 240s irradiation, only 0.01% (±0.01%) of E. faecalis cells and 0.03% (±0.03%) of F. nucleatum survived after treatment. This study indicated that RB can bind to E. faecalis and F. nucleatum in a sufficient amount to elicit effective aPDT. Epifluorescence microscopy showed a yet-unreported property of RB binding to bacterial membranes. Flow cytometry allowed the detection of bacteria with damaged membranes that were unable to form colonies on agars after cell sorting.

Repeated exposures to blue light-activated eosin Y enhance inactivation of E. faecalis biofilms, in vitro.

BACKGROUND: In dentistry, antibacterial photodynamic therapy (a-PDT) has shown promising results for inactivating bacterial biofilms causing carious, endodontic and periodontal diseases. In the current study, we assessed the ability of eosin Y exposed to 3 irradiation protocols at inactivating Enterococcus faecalis biofilms, in vitro. METHODS: E. faecalis biofilms formed on hydroxyapatite disks were incubated with eosin Y (10-80µM), then activated with blue light using different irradiation protocols. Biofilms exposed to continuous exposure were incubated for 40min before being light-activated for 960 s. For the intermittent exposure, biofilms were exposed 4 times to the light/photosensitizer combination (960 s total) without renewing the photosensitizer. For repeated a-PDT, the same light dose was delivered in a series of 4 irradiation periods separated by dark periods; fresh photosensitizer was added between each light irradiation. After treatment, bacteria were immediately labeled with LIVE/DEAD BacLight Bacterial Viability kit and viability was assessed by flow cytometry (FCM). Results were statistically analyzed using one-way ANOVA and Tukey multiple comparison intervals (α=0.05). RESULTS: The viability of E. faecalis biofilms exposed to 10µM eosin Y, was significantly reduced compared to controls (light only-eosin Y only). After a second exposure to blue light-activated eosin Y, viability significantly decreased from 58% to 12% whereas 6.5% of the bacterial biofilm remained live after a third exposure (p<0.05). Only 3.5% of the bacterial population survived after the fourth exposure. CONCLUSIONS: The results of this study indicate that blue light-activated eosin Y can photoinactivate E. faecalis biofilms grown on hydroxyapatite disks. Also, repeated exposures to blue light-activated eosin Y were shown to significantly improve efficacy. Further studies seem warranted to optimize the antibacterial activity of blue light-activated eosin Y on major oral pathogens.

Flow cytometric assessment of Streptococcus mutans viability after exposure to blue light-activated curcumin.

BACKGROUND: Streptococcus mutans biofilms are considered as primary causative agents of dental caries. Photodynamic antimicrobial chemotherapy (PACT) has been recently proposed as a strategy for inactivating dental biofilms. This study aimed to investigate the effect of blue light-activated curcumin on S. mutans viability and to explore its potential as a new anti-caries therapeutic agent. The effect of different concentrations and incubation times of photo-activated curcumin on the survival of S. mutans in planktonic and biofilm models of growth was assessed by flow cytometry. METHODS: Streptococcus mutans in planktonic suspensions or biofilms formed on hydroxyapatite disks were incubated for 5 or 10min with curcumin prior to blue light activation. Bacteria were labeled with SYTO 9 and propidium iodide before viability was assessed by flow cytometry. Results were statistically analyzed using one-way ANOVA and Tukey multiple comparison intervals (α=0.05). RESULTS: For planktonic cultures, 0.2µM of light-activated curcumin significantly reduced S. mutans viability (p<0.05). For biofilm cultures, light-activated curcumin at concentration of 40-60µM only suppressed viability by 50% (p<0.05). Independently of the mode of growth, incubation time has no significant effect on PACT efficiency. CONCLUSION: This study indicates that blue light-activated curcumin can efficiently inactivate planktonic cultures of S. mutans whereas biofilms were more resistant to treatment. Flow cytometry allowed the detection of bacteria with damaged membranes that were unable to replicate and grow after cell sorting. Further studies seem warranted to optimize the efficacy of light-activated curcumin against S. mutans biofilms.

Microhardness of light- and dual-polymerizable luting resins polymerized through 7.5-mm-thick endocrowns.

STATEMENT OF THE PROBLEM: The complete polymerization of luting resins through thick indirect restorations is still questioned. PURPOSE: The purpose of this study was to evaluate the degree of polymerization of light- and dual-polymerizable luting resins under thick indirect composite resin and ceramic endocrowns by means of Vickers microhardness measurements. MATERIAL AND METHODS: The Vickers microhardness measurements of a light-polymerizable microhybrid composite resin and a dual-polymerizable luting cement directly polymerized in a natural tooth mold for 40 seconds with a high-power light-emitting diode lamp (control) were compared with measurements after indirect irradiation through 7.5-mm-thick composite resin and ceramic endocrowns for 3 × 90 seconds. A test-to-control microhardness values ratio of 0.80 at a depth of 0.5 mm below the surface was assumed as the criterion for adequate conversion. RESULTS: For the Vickers microhardness measurements of a dual-polymerizable luting cement, no differences (P>.05) were found between Vickers microhardness control values and values reported after polymerization through composite resin and ceramic endocrowns. For The Vickers microhardness measurements (±SD) of a light-polymerizable microhybrid composite resin, control values were significantly (P<.05) higher (111 ±3.3) than those reported after polymerization through composite resin (100.5 ±3.8) and ceramic (99.7 ±2.3) endocrowns. However, the hardness values of The Vickers microhardness measurements of a light-polymerizable microhybrid composite resin polymerized through the endocrowns were approximately 10% to 12% lower than those of the control values. Two-way ANOVA showed the influence of the luting material on the Vickers microhardness values (P<.05). The effect of endocrown material was not significant (P>.05). CONCLUSIONS: Under the conditions of this in vitro study, Vickers microhardness values of the dual-polymerizable resin cement and the light-polymerizable restorative composite resin irradiated for 3 × 90 seconds with a high irradiance light-emitting diode lamp through 7.5-mm-thick endocrowns reached at least 80% of the control Vickers microhardness values, which means that both materials can be adequately polymerized when they are used for luting thick indirect restorations.

Antimicrobial activity and cytotoxicity of 3 photosensitizers activated with blue light.

INTRODUCTION: Pulp repair is less likely to occur when dentin or pulpal tissue remains infected after caries excavation. Yet there are currently few options to kill residual bacteria without damaging resident cells. The current study has evaluated the effect of 3 blue light-activated chemicals on the viability of lactobacilli, odontoblast-like cells (MDPC-23), undifferentiated pulp cells (OD21), and human embryonic stem cells (hESC H1). METHODS: Bacteria were incubated for 15 minutes with curcumin, eosin Y, or rose bengal and then irradiated with blue light (240 seconds). Bacteria were labeled with LIVE/DEAD BacLight Bacterial Viability kit, and viability was assessed by fluorescence-activated cell sorting. Cytotoxicity assays were performed on MDPC-23 cells, OD21, and hESC H1 cells grown in 24-well plates and exposed to the same photosensitizer-light combination. After 24 hours, cellular response was measured by using the methyl-thiazol-diphenyl-tetrazolium assay. Results were statistically analyzed by using one-way analysis of variance and Tukey multiple comparison intervals (α = 0.05). RESULTS: Bacterial viability was significantly reduced after exposure to different combinations of light and photosensitizers; mitochondrial activity of cultured cells remained unaffected when exposed to the same conditions, suggesting a good therapeutic index in vitro. CONCLUSIONS: Blue light-mediated disinfection is promising for the development of new treatment strategies designed to promote pulp repair after carious exposure.

Blue light-mediated inactivation of Enterococcus faecalis in vitro.

In dentistry, residual infection remains a major cause of failure after endodontic treatment; many of these infections involve Enterococcus faecalis. In the current study, we explored the possibility that blue light activated photosensitizers could be used, in principle, to inactivate this microbe as an adjunct disinfection strategy for endodontic therapy. Three blue light absorbing photosensitizers, eosin-Y, rose bengal, and curcumin, were tested on E. faecalis grown in planktonic suspensions or biofilms. Photosensitizers were incubated for 30 min with bacteria then exposed to blue light (450-500 nm) for 240 s. Sodium hypochlorite (3%) was used as a control. After 48 h, the viability of E. faecalis was estimated by measuring colony-forming units post-exposure vs. untreated controls (CFU/mL). Blue light irradiation alone did not alter E. faecalis viability. For planktonic cultures, blue light activated eosin-Y (5 µM), rose bengal (1 µM), or curcumin (5 µM) significantly (p<0.05) reduced E. faecalis viability compared to exposure to the unirradiated photochemicals. For biofilm cultures, concentrations of light-activated eosin-Y, rose bengal, and curcumin of 100, 10, and 10 µM respectively, completely suppressed E. faecalis viability (p<0.05). Although the current results are limited to an in vitro model, they support further exploration of blue light activated antimicrobials as an adjunct therapy in endodontic treatment.

Cytotoxic response of three cell lines exposed in vitro to dental endodontic sealers.

The in vitro cytotoxicity of five endodontic sealers was measured >8-12 weeks using L929 mouse fibroblasts, osteoblastic cells (ROS) 17/2.8 rat osteoblasts, and MC3T3-E1 mouse osteoblasts. Discs (n = 6) of AH-plus Jet (AHP), two versions of Endo Rez (ER, ERx), Epiphany (EPH), and Pulp Canal Sealer (PCS) were prepared. The sealers and Teflon (Tf, negative control) were placed in direct contact with cells after immersion in phosphate-buffered saline for 1-12 wk. Cellular succinate dehydrogenase (SDH) activity was estimated using the MTT method (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, a yellow tetrazole), and activities were normalized to Teflon® controls. The cellular responses to the materials were compared using analysis of variance with Tukey posthoc analyses (α = 0.05). Initially, all sealers suppressed normalized SDH activity of L929 fibroblasts by >90%. After 12 weeks of immersion in saline, AHP exhibited the SDH activity above Tf (120%), followed by ERx (78%), ER (58%), PCS (38%), and EPH (28%), all statistically distinct (p < 0.05). In general, the three cell lines responded similarly to the sealers. However, AHP caused unique responses: ROS cells were significantly (p < 0.05) less sensitive initially, and AHP was severely cytotoxic to MC3T3 cells (<35% of Tf) through 8 weeks. The data suggest that with "aging" in saline, current endodontic sealers decrease in in vitro cytotoxicity at different rates.

Production of reactive oxygen species from photosensitizers activated with visible light sources available in dental offices.

OBJECTIVES: The aim of this study was to assess the ability of commonly available red- or blue-light dental sources to generate reactive oxygen species (ROS) from photosensitive chemicals that might be useful for photodynamic antimicrobial chemotherapy (PACT). BACKGROUND: Although the use of red diode lasers is well documented, there is limited information on how useful blue-light sources might be for PACT in dental contexts. MATERIALS AND METHODS: A diode laser (Periowave; see Table 1 for material and equipment sources) emitting red light (660-675 nm) was used to activate toluidine blue; riboflavin and pheophorbide-a polylysine (pheophorbide-a-PLL) were photoactivated using an Optilux 501 curing unit emitting blue light (380-500 nm). Ozone gas (generated by OzoTop, Tip Top Tips, Rolle, Switzerland), sodium hypochlorite, and hydrogen peroxide were used for comparison. ROS production was estimated using an iodine-triiodide colorimetric assay, and ROS levels were plotted versus concentration of chemicals to determine each chemical's efficiency in ROS production. One-way ANOVA with Tukey post hoc analysis (alpha = 0.05) was used to compare the efficiencies of ROS production for the various chemicals. RESULTS: Sodium hypochlorite, hydrogen peroxide, and ozone gas produced ROS spontaneously, whereas pheophorbide-a-PLL, riboflavin, and toluidine blue required light exposure. The efficiency of ROS production was higher for pheophorbide-a-PLL and toluidine blue than for ozone gas or riboflavin (p < 0.05). Hydrogen peroxide was the least efficient ROS producer. CONCLUSIONS: The results of the current study support the use of blue- or red-light-absorbing photosensitizers as candidates to produce ROS for clinical applications. Blue-light photosensitizers were as efficient as red-light photosensitizers in producing ROS and more efficient than the oxidant chemicals currently used for dental disinfection.

Inflammatory suppression by endodontic sealers after aging 12 weeks In vitro.

Dental endodontic sealers are in intimate contact with tissues around the root apex (periapical area) for extended periods. New endodontic sealers have been developed in the past decade, but the biological responses to many new products are not well documented. In this study, we assessed in vitro monocytic cytotoxic and inflammatory responses to several contemporary endodontic sealers. AH-Plus (AH), Pulp Canal Sealer (PC), Epiphany (EPH), Endo-Rez (ER), and an experimental Endo-Rez (ERx) were initially placed in buffered-saline for 12 weeks to simulate in vivo use. After "aging," specimens were placed in direct contact with THP1 monocytes for 72 h and their cytotoxicity (mitochondrial response; MTT) or ability to trigger or suppress cytokine secretion (ELISA; TNFalpha, IL1beta, IL=6; +/- lipopolysaccharide (LPS) exposure) were measured relative to Teflon (Tf) negative controls. Cellular responses among conditions were compared with ANOVA and Tukey post-hoc analysis (alpha = 0.05). Two of the five sealers, EPH and PC, still suppressed cell mitochondrial activity by 70% or more after 12 weeks of conditioning in saline. No sealer alone activated monocytic TNFalpha, IL1beta, or IL6 secretion (p > 0.05 vs. +LPS controls). When THP1 were activated by LPS after exposure to the sealers, differential suppression of TNFalpha, IL1beta, and IL6 secretion was observed for two of the five sealers tested. (EPH and PC) This data suggest that common endodontic sealers do not activate monocytic TNFalpha, IL1beta, and IL6 secretion in vitro by themselves, but degradation products of the sealers may suppress activation of monocytes.

Accuracy of electronic apex locater-controlled handpieces.

An in vitro model was used to test a hypothesis that the manual (apex locator) and motor-driven (apex locator-controlled handpiece) operating modes of 3 newly developed apex-locator-controlled handpiece devices (Dentaport ZX, Endomaster, XSmart Dual) give the same working length. The depth of penetration of the file into the root canal was measured using a digital micrometer and the distance of the tip of the file relative to the major root foramen was determined by scanning electron microscopy (SEM). In the manual mode, the XSmart Dual device reported significantly shorter working lengths than the Dentaport ZX or the Endomaster devices. In the motor-driven mode, the XSmart Dual device reported significantly longer working lengths than the Dentaport ZX but not the Endomaster. Most instruments driven by the handpieces were confined within the root canal and differences in working lengths between manual and motor-driven modes were small for all devices (tenths of millimeters). We concluded that although the motor-driven mode of these devices appeared to be clinically safe, measurements obtained in manual and motor-driven operating modes are not equivalent.

Intracellular reactive oxygen species in monocytes generated by photosensitive chromophores activated with blue light.

OBJECTIVES: Disinfection of the tooth pulp-canal system is imperative to successful endodontic therapy. Yet, studies suggest that 30-50% of current endodontic treatments fail from residual bacterial infection. Photodynamic therapy using red-light chromophores (630 nm) to induce antimicrobial death mediated by generated reactive oxygen species (ROS) has been reported, but red-light also may thermally damage resident tissues. In the current study, we tested the hypothesis that several blue light chromophores (380-500 nm) generate intracellular reactive oxygen species but are not cytotoxic to mammalian cells. METHODS: THP1 monocytes were exposed to 10 microM of four chromophores (chlorin e6, pheophorbide-a, pheophorbide-a-PLL, and riboflavin) for 30 min before activation with blue light (27J/cm(2), 60s). After activation, intracellular ROS were measured using a dihydrofluorescein diacetate technique, and cytotoxicity was determined by measuring mitochondrial activity with the MTT method. RESULTS: All photosensitizers produced intracellular ROS levels that were dependent on both the presence of the photosensitizer and blue light exposure. Riboflavin and pheophorbide-a-PLL produced the highest levels of ROS. Photosensitizers except riboflavin exhibited cytotoxicity above 10 microM, and all except pheophorbide-a-PLL were more cytotoxic after blue light irradiation. SIGNIFICANCE: The current study demonstrated the possible utility of blue light chromophores as producers of ROS that would be useful for endodontic disinfection.

In vitro cytotoxic response to lithium disilicate dental ceramics.

OBJECTIVES: The use of lithium disilicate dental ceramics is increasing in dentistry and previous reports have suggested that they may have greater biological risks than previously thought. We tested a hypothesis that composition and processing influence the biological properties of these ceramics. METHODS: The cytotoxicity of two machined and three pressed lithium disilicate materials (n=6) were tested in vitro using mouse fibroblasts in direct contact with the materials for 72h. Cellular response was estimated by mitochondrial succinate dehydrogenase activity (MTT method). Mitochondrial activity was expressed as a percentage of Teflon controls, then compared to Teflon using 2-sided t-tests (alpha=0.05). Polished materials were aged in artificial saliva and tested for cytotoxicity periodically over 6 weeks, then were repolished (320grit SiC paper), aged and tested again for 4 weeks. RESULTS: All materials significantly (50-70%) suppressed cellular mitochondrial activity in the initial week, but suppression decreased by 25-30% over the next 2 weeks. In weeks 4 and 6 some materials exhibited a cytotoxic 'relapse' of 10-20%. The cytotoxic response was no different for machined or pressed materials, but the presence of ZnO had at least an association with longer-term cytotoxicity and relapse. Repolishing to 320grit did not increase cytotoxicity significantly. SIGNIFICANCE: Our results suggest that lithium disilicates are not biologically inert, and that many have a similar cytotoxicity dynamic regardless of small differences in composition or processing.

Effect of cyclic loading under enzymatic activity on resin-dentin interfaces of two self-etching adhesives.

OBJECTIVES: To evaluate microtensile bond strength and micro-morphology at the resin-dentin interfaces of two self-etching adhesive systems subjected to simultaneous mechanical and enzymatic stress. METHODS: Sixteen enamel/dentin discs were bonded with a two-step self-etching adhesive (AdheSE, n=8) and a one-step self-etching adhesive (Xeno III, n=8) to a 2mm thick resin composite layer. One resin-dentin bar was obtained per tooth. In half of the specimens of each group microTBS and micro-morphological evaluations (TEM) was performed without loading. The other half was mechanically loaded in a cholinesterase-containing solution. MicroTBS as well as ultra-morphological evaluations of the directly loaded areas using TEM were performed on the loaded specimens. RESULTS: The microTBS of the specimens (non-loaded/loaded) were of 39.6+/-14.7/35.4+/-22.1 and of 21.8+/-29.8/15.9+/-25.5 for AdheSE and Xeno III, respectively. Under TEM, both materials presented signs of nanoleakage. However, on loaded specimens the extent of nanoleakage was slightly reduced for AdheSE and no silver staining was observed on the adhesive interface of Xeno III. TEM evaluations of the specimens' loaded area revealed no decrease in the width of the adhesive interface for AdheSE. The contrary was observed in the interface created by Xeno III. SIGNIFICANCE: The adhesive interfaces created by the two-step self-etching adhesive (AdheSE) could better withstand both mechanical and enzymatic stresses on the long-term than the one-step self-etching system (Xeno III) tested in the present study.

Alternative adhesive strategies to optimize bonding to radicular dentin.

This study tested the hypothesis that bond strengths of filling materials to radicular dentin might be optimized by using an indirect dentin bonding procedure with an acrylic core material. Roots of human teeth were endodontically prepared and obturated with EndoREZ, Epiphany, or the bonding of an acrylic point with SE Bond by using a direct or an indirect bonding technique. Bond strengths of endodontic sealers to radicular dentin were measured with a thin slice push-out test. Push-out strengths of EndoREZ and Epiphany to radicular dentin were less than 5 megapascals (MPa). The direct bonding technique with acrylic points and the self-etching adhesive had push-out strengths of 10 MPa, increasing to 18 MPa with the indirect technique. The use of the indirect bonding protocol with an acrylic point to compensate for polymerization stresses appears to be a viable means for optimizing bond strengths of endodontic filling materials to radicular dentin.

Push-out bond strengths of endodontic posts bonded with different resin-based luting cements.

PURPOSE: To compare the push-out bond strengths of endodontic posts bonded with different resin-based luting cements and to verify that bond strengths did not vary with cement thickness. METHODS: 48 root canals were shaped using 6% NiTi rotary files, obturated with gutta-percha and AH Plus sealer and prepared for post cementation using Panavia F, Parapost cement, SuperBond and Unicem Rely X. All roots were sectioned into 0.7 mm thick slices and digital photographs of each slice were analyzed using Scion Image to measure the surface area of the luting cement. The root slices were stressed to failure at 1 mm/minute using a push-out test. Push-out strength was calculated as the force at failure divided by the bonded surface area. Least squares linear regression analysis was used to assess the effect of cement thickness on bond strength. Fractured specimens were further observed under the SEM. RESULTS: Mean push-out bond strengths were: Panavia F (8.8 +/- 3.6 MPa), Parapost cement (9.1 +/- 4.4 MPa) SuperBond (14.6 +/- 2.9 MPa) and Rely X Unicem (12.4 +/- 3.3 MPa). The Panavia F and the Parapost cement were not significantly different from each other, but both were significantly lower (P < or = 0.05) than SuperBond and Rely X Unicem. Although there were large variations in cement thickness, the cementation of fiber posts with thicker cement layers did not affect the performance of the adhesive luting cements applied to root canal dentin.

Influence of different surface treatments on marginal adaptation in enamel and dentin.

PURPOSE: To compare marginal adaptation in enamel and dentin after different surface treatments before and after long-term simultaneous thermal and mechanical stresses in a mixed Class V restoration. MATERIALS AND METHODS: Thirty-six V-shaped mixed Class V cavities were prepared in extracted human molars and treated as follows: group 1: 30 s ozone exposure (Heal Ozone, Kavo); group 2: 20 s air abrasion with 50 microm Al2O3 particles (Dento-prep, Rønvig); group 3: 20 s exposure to 27 microm SiOx powder (RONDOflex, Kavo with CoJet powder, 3M-ESPE); group 4: control (no treatment). Cavities were restored with a light-cured composite material (Tetric Ceram, shade A2, Ivoclar Vivadent) using a self-etching adhesive system (Syntac Clasic, Ivoclar Vivadent) with H3PO4 conditioning of the enamel. Each group was evaluated in respect to marginal adaptation before and after mechanical and thermal loading under simulated dentinal fluid. RESULTS: Even if loading significantly influenced marginal quality in all groups (paired t-test, p < 0.05), the percentages of "continuous margin" of all groups in enamel ranged between 93.2% and 92.3% before and 84.1% and 76.9% after loading and were not significantly different (ANOVA and Scheffe's post-hoc test, p > 0.05). Continuous margin in dentin ranged from 98.9% to 94.2% before and from 95.9% to 76.4% after loading, and significant differences were observed between groups treated with ozone vs control before and after loading and CoJet vs control group after loading (ANOVA and Scheffe's post-hoc test, p < 0.05). CONCLUSION: Surface treatment with ozone and silica coating may significantly decrease marginal quality in dentin without negatively influencing marginal quality in enamel.

Initial in vitro biological response to contemporary endodontic sealers.

The objective of this study was to evaluate the cytotoxicity of three endodontic sealers (AH Plus/Maillefer-Dentsply, Epiphany/Pentron, GuttaFlow, Coltene-Whaledent). Materials were mixed according to the manufacturer instructions and packed into Teflon molds (10 x 1 mm). For cytotoxicity testing (MTT method), the specimens were placed in contact with cultured cells, then evaluated at two subsequent time points (24 or 72 h). In addition to testing the mixed materials, 5 microl of primer liquid (GuttaFlow and Epiphany) and resin solvents (HEMA, ethanol, sterile water, or acetone) were added directly in culture for 24 and 72 h. The results showed that most materials pose significant cytotoxic risks and that cytotoxicity generally increased with time. At 72 h, GuttaFlow became significantly less toxic than AH Plus, Epiphany sealer, and Resilon. The current results support the need to continue to develop better endodontic sealers that combine the excellent sealing and bonding properties of resins with acceptable biological properties for endodontic applications.