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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

Sublethal concentrations of diverse gold compounds inhibit mammalian cytosolic thioredoxin reductase (TrxR1).

UNLABELLED: Thioredoxin reductase (TrxR) reduces thioredoxin (Trx), thereby contributing to cellular redox balance, facilitating the synthesis of deoxy-ribose sugars for DNA synthesis, and regulating redox-sensitive gene expression. Auranofin is a gold compound that potently inhibits TrxR. This inhibition is one suspected mechanism of auranofin's therapeutic benefit in the treatment of rheumatoid arthritis. The use of other gold compounds to treat cancer or inflammatory disease may rely on their ability to inhibit TrxR. In the current study, we tested the hypothesis that a variety of gold compounds may inhibit TrxR. METHODS: We exposed rat-TrxR1 to auranofin, gold sodium thiomalate, sodium aurothiosulfate, triphenyl phosphine gold chloride, or gold acetate, and measured TrxR activity ex vivo. We then compared TrxR1 inhibitory levels of gold compounds to those that inhibited mitochondrial activity of THP1 monocytes and OSC2 epithelial cells, estimated by succinate dehydrogenase activity. RESULTS: All gold compounds inhibited TrxR1 at concentrations ranging from 5 to 4000 nM (50% inhibitory concentration). The oxidation state of gold did not correlate with inhibitory potency, but ligand configuration was important. Au(I)-phosphine compounds (triphenyl phosphine gold chloride and auranofin) were the most potent inhibitors of TrxR. All TrxR1 inhibitory concentrations were sublethal to mitochondrial activity in both THP1 and OSC2 cells. CONCLUSIONS: Diverse types of gold compounds may be effective inhibitors of TrxR1 at concentrations that do not suppress cellular mitochondrial function. Inhibition may be optimized to some degree by altering the ligand configuration of the compounds. These results support future study of a variety of Au compounds for therapeutic development as inhibitors of TrxR1.

Gold-induced reactive oxygen species (ROS) do not mediate suppression of monocytic mitochondrial or secretory function.

UNLABELLED: The toxicity of anti-rheumatic gold compounds has limited their use and development, yet both the toxicological and therapeutic actions of these compounds remain unclear. In the current study, we tested the hypothesis that intracellular reactive oxygen species (ROS) induced by Au(I) or Au(III) compounds mediate their ability to suppress mitochondrial activity. METHODS: Human THP1 monocytes were exposed to HAuCl(4) x 3H(2)O (Au(III)), or the anti-rheumatic compounds auranofin (AF) or gold sodium thiomalate (GSTM) for 6-72 h, after which mitochondrial activity (succinate dehydrogenase) was measured. To assess the role of cellular redox status as a mediator of mitochondrial suppression, monocytes were pre-treated with a pro-oxidant (t-butyl hydroquinone, t-BHQ) or antioxidant (N-acetyl cysteine, NAC ). ROS levels were measured 0-24h post-gold addition to determine their role as mediators of mitochondrial activity suppression. RESULTS: AF was the most potent inhibitor of mitochondrial activity, followed by Au(III) and GSTM. Only Au(III) induced intracellular ROS; no ROS formation was observed in response to AF or GSTM exposure. Although anti- and pro-oxidants had some effects on mitochondrial suppression of Au compounds, collectively the data do not support redox effects or ROS formation as major mediators of Au-compound mitochondrial suppression. CONCLUSIONS: Our results do not indicate that ROS and redox effects play major roles in mediating the cytotoxicity of AF, GSTM or Au(III).

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.

Hydrothermal and mechanical stresses degrade fiber-matrix interfacial bond strength in dental fiber-reinforced composites.

Fiber-reinforced composites (FRCs) show great promise as long-term restorative materials in dentistry and medicine. Recent evidence indicates that these materials degrade in vivo, but the mechanisms are unclear. The objective of this study was to investigate mechanisms of deterioration of glass fiber-polymer matrix bond strengths in dental fiber-reinforced composites during hydrothermal and mechanical aging. Conventional three-point bending tests on dental FRCs were used to assess flexural strengths and moduli. Micro push-out tests were used to measure glass fiber-polymer matrix bond strengths, and nanoindentation tests were used to determine the modulus of elasticity of fiber and polymer matrix phases separately. Bar-shaped specimens of FRCs (EverStick, StickTech, and Vectris Pontic, Ivoclar-Vivadent) were either stored at room temperature, in water (37 and 100 degrees C) or subjected to ageing (10(6) cycles, load: 49 N), then tested by three-point bending. Thin slices were prepared for micro push-out and nanoindentation tests. The ultimate flexural strengths of both FRCs were significantly reduced after aging (p < 0.05). Both water storage and mechanical loading reduced the interfacial bond strengths of glass fibers to polymer matrices. Nanoindentation tests revealed a slight reduction in the elastic modulus of the EverStick and Vectris Pontic polymer matrix after water storage. Mechanical properties of FRC materials degrade primarily by a loss of interfacial bond strength between the glass and resin phases. This degradation is detectable by micro push-out and nanoindentation methods.

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.

Thermal risks from LED- and high-intensity QTH-curing units during polymerization of dental resins.

The aim of this study was to test the ability of an infrared (IR) camera to assess temperature changes and distributions in teeth below restorations when quartz-tungsten-halogen (QTH) and light-emitting diode (LED) curing lights were used to photopolymerize the restorative material. Our hypothesis was that the higher power density and broader spectral distribution of the QTH source would cause greater increases in tooth temperature than the LED source, and that these differences would be best demonstrated with the IR camera. Cavities were prepared on human third molars and restored with a resin composite restorative material. The material was light-cured using three light-curing sources using several exposure times. The external (outside the tooth) and internal (inside the pulp chamber) temperature changes during polymerization of the composite material were recorded over 360 s with thermocouples and an IR camera. Using thermocouples the maximum increase in external temperature (+17.7 degrees C) was reported for the Swiss Master light after 20 s of curing time while the minimum temperature rise (+7.8 degrees C) was reported for the Freelight 2. Whereas a 2.6 degrees C increase in internal temperature was observed after curing 20 s with the Freelight 2, 7.1 degrees C was reported after 60 s of light exposure to Astralis 10. Infrared images showed similar trends in external-internal rises in temperature as the thermocouples, although temperatures measured by the IR were generally higher. These results indicate that the higher power density QTH sources caused greater increases in tooth temperatures than the LED source and that thermocouples may underestimate the heat applied to the tooth.

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.

Toothbrushing causes elemental release from dental casting alloys over extended intervals.

The release of elements from dental alloys has been linked to alloy biocompatibility. Much of the research measuring elemental release has been done in vitro under passive conditions. The current study supplements a previous report that measured elemental release from dental alloys during and after the equivalent of 1 week of toothbrushing. In the current study, toothbrushing times were extended to the equivalent of 2 years, and elemental release was measured during and after brushing, with and without toothpaste. The results showed that for the major classes of dental alloys, brushing alone caused no significant elemental release during the brushing, and only minor increases after brushing. Brushing with toothpaste caused significant increases in elemental release for all elements of all alloys, but the largest increases were for the two nickel-based alloys. Nickel released during brushing with toothpaste reached 600-800 microg/cm(2) of alloy surface. Both beryllium-containing and non-beryllium-containing nickel-based alloys behaved similarly, refuting claims that non-beryllium alloys are superior in this regard. Thus, brushing with toothpaste under these extended in vitro conditions appears to increase the biological liabilities from elemental release for all alloys, but primarily for nickel-based alloys.

Sublethal concentrations of Au (III), Pd (II), and Ni(II) differentially alter inflammatory cytokine secretion from activated monocytes.

Many transition metals have been viewed collectively as nonspecific biological toxins in cells, which has limited investigation into their possible therapeutic effects. In the current study, the effects of Au(III), Ni(II), and Pd(II) on the differential secretion of cytokines from monocytes has been investigated. This is critical to understanding any therapeutic potential of these metals, their allergenicity, or the clinical effects of current metal therapies such as chrysotherapy. Lethal concentrations (defined as > 50% suppression of mitochondrial succinate dehydrogenase (SDH) activity) of metals were determined by dose-response curves with the use of 72 h exposures to human THP-1 monocytes. Then, secretion of TNFalpha, IL1beta, and IL6 were measured after the monocytes were exposed to sublethal concentrations of metals, with or without stimulation by lipopolysaccharide. The concentrations of Au(III), Pd(II), and Ni(II) required to suppress SDH activity by 50% were found to be 255, 270, and 90 microM, respectively. No sublethal concentration of any metal alone caused secretion of the cytokines. However, LPS-induced cytokine secretion was significantly and differentially altered by sublethal concentrations of each metal. Differential responses were highly dependent on metal concentration and involved both suppression and potentiation of the LPS activation. In the case of Ni(II), potentiation of TNFalpha, IL1beta, and IL6 ranged from 200% for TNFalpha to over 1200% for IL6. Metals such as Au(III), Pd(II), and Ni(II) differentially alter cytokine expression from monocytes. These results imply that metals have more specific effects on cell signaling than previously assumed. These results also are important in explaining multiple clinical effects often seen with chrysotherapy, identifying potential new avenues for metal therapy, and understanding the inflammatory effects of metals such as nickel.

In vitro biological response to core and flowable dental restorative materials.

OBJECTIVES: In vitro cytotoxicities of commercially available core and flowable dental restorative materials were assessed and compared to traditional resin composites. Our hypothesis was that the increased resin diluents added to achieve higher flow in flowables would increase cytotoxicities, whereas the higher filler content of core materials would decrease cytotoxicities relative to traditional resin composites. METHODS: Specimens were made under aseptic conditions, then extracted into an artificial saliva solution for 0-4 weeks, to assess the effect of aging on cytotoxicity. After extraction, specimens were tested for cytotoxicity in vitro using Balb/c fibroblasts in direct contact format. Cells were exposed to the materials for 48h, after which the mitochondrial activity of the cells was measured (MTT method). Cellular activity was normalized to Teflon negative controls. RESULTS: Core materials were uniformly and severely (<50% of Teflon cellular activity) cytotoxic initially, but several materials (Corepaste, Definite core) improved somewhat with aging in artificial saliva. Flowable materials were uniformly and severely cytotoxic with no trend toward improvement with aging. The Definite-flow was the least cytotoxic of the flowable materials, but it too was severely cytotoxic. SIGNIFICANCE: Commercially available core and flowable restorative materials showed severe in vitro cytotoxicities that are worse than some traditional composites and most dental casting alloys and amalgams used today. Of particular note was the persistent cytotoxicity of these materials after 4 weeks of extraction with artificial saliva. These cytotoxicities indicate a continuing release of mass from these materials at levels that have biological relevance in vitro. In vivo relevance of these cytotoxicities is less clear, but these results indicate a higher biological risk for these materials compared to traditional materials that exhibit less initial toxicity and improve with aging time.