Titanium dioxide nanotubes in a hydrogen peroxide-based bleaching agent: physicochemical properties and effectiveness of dental bleaching under the influence of a poliwave led light activation.

OBJECTIVES: The effects of different concentrations of titanium dioxide (TiO2) into 40% hydrogen peroxide (HP) were evaluated as regards the effectiveness of dental color change either associated with activation by polywave LED light or not. MATERIALS AND METHODS: TiO2 (0, 1, 5, or 10%) was incorporated into HP to be applied during in-office bleaching (3 sessions/40 min each). Polywave LED light (Valo Corded/Ultradent) was applied or not in activation cycles of 15 s (total time of 2 min). The color of 80 third molars separated into groups according to TiO2 concentration and light activation (n = 10) was evaluated at baseline and at time intervals after the 1st, 2nd, and 3rd bleaching sessions. RESULTS: WID value was significantly higher when using HP with 5% TiO2 in the 2nd session than the values in the other groups (p < 0.05). After the 2nd and 3rd sessions, the ΔEab value was significantly higher when activated with light (p < 0.05) for all agents containing TiO2 or not. Zeta potential and pH of the agents were not modified by incorporating TiO2 at the different concentrations. CONCLUSIONS: The 5% TiO2 in the bleaching agent could enhance tooth bleaching, even without light application. Association with polywave LED light potentiated the color change, irrespective of the presence of TiO2 in the bleaching gel. CLINICAL SIGNIFICANCE: HP with 5% TiO2 could lead to a greater tooth bleaching response in the 2nd clinical session, as well as the polywave light can enhance color change.

TiO2 nanotube-based nanotechnology applied to high-viscosity conventional glass-ionomer cement: ultrastructural analyses and physicochemical characterization.

This study characterized TiO2 nanotube (TiO2-nt) ultrastructure and morphology, and the physicochemical impact on high-viscosity conventional glass-ionomer cement (GIC). TiO2-nt was synthesized by the alkaline method (n = 3), assessed by scanning (SEM) and transmission electron microscope (TEM), and was added (3%, 5%, 7%-in weight) to KM (Ketac Molar EasyMix™). Analyses included: SEM; Energy-dispersive spectroscopy (EDS); Raman spectroscopy (RAMAN); Setting time with Gillmore needles (ST); Color (Co); Radiopacity (XR); Water sorption (WS); and solubility (SO). Quantitative data were submitted to ANOVA and Tukey's tests (chr = 0.05). External and internal TiO2-nt diameters were 11 ± 2 nm and 6 ± 0 nm, respectively. Data analyses showed: (i) TiO2-nt present into KM matrix, with a concentration-dependent increase of Ti levels into KM, (ii) physical interaction between KM and TiO2-nt, (iii) longer initial ST for the 7% group compared to KM and 3% groups (p ≤ 0.01), (iv) decreased luminosity and yellowness for the 5% and 7% groups, (v) 36% greater radiopacity for the 5% group compared to enamel, dentin, and KM, and (vi) lower SO values for the 5% group, with no significant differences on WS across the groups. TiO2-nt displayed physical interaction with KM matrix, and also modified SO, XR and Co, without affecting ST. This study provides information on the potential impact of TiO2-nt on GIC performance. TiO2-nt may be proposed to boost confidence among dental surgeons in terms of GIC's handling characteristics, success rate and differential diagnostic.

Novel LRAP-binding partner revealing the plasminogen activation system as a regulator of cementoblast differentiation and mineral nodule formation in vitro.

Amelogenin isoforms, including full-length amelogenin (AMEL) and leucine-rich amelogenin peptide (LRAP), are major components of the enamel matrix, and are considered as signaling molecules in epithelial-mesenchymal interactions regulating tooth development and periodontal regeneration. Nevertheless, the molecular mechanisms involved are still poorly understood. The aim of the present study was to identify novel binding partners for amelogenin isoforms in the cementoblast (OCCM-30), using an affinity purification assay (GST pull-down) followed by mass spectrometry and immunoblotting. Protein-protein interaction analysis for AMEL and LRAP evidenced the plasminogen activation system (PAS) as a potential player regulating OCCM-30 response to amelogenin isoforms. For functional assays, PAS was either activated (plasmin) or inhibited (ε-aminocaproic acid [aminocaproic]) in OCCM-30 cells and the cell morphology, mineral nodule formation, and gene expression were assessed. PAS inhibition (EACA 100 mM) dramatically decreased mineral nodule formation and expression of OCCM-30 differentiation markers, including osteocalcin (Bglap), bone sialoprotein (Ibsp), osteopontin (Spp1), tissue-nonspecific alkaline phosphatase (Alpl) and collagen type I (Col1a1), and had no effect on runt-related transcription factor 2 (Runx2) and Osterix (Osx) mRNA levels. PAS activation (plasmin 5 µg/µl) significantly increased Col1a1 and decreased Bglap mRNA levels (p < .05). Together, our findings shed new light on the potential role of plasminogen signaling pathway in the control of the amelogenin isoform-mediated response in cementoblasts and provide new insights into the development of targeted therapies.

Shifts in the bacterial community of saliva give insights on the relationship between obesity and oral microbiota in adolescents.

The current study aimed at the determination of the impact of obesity on the salivary microbiome in adolescents. Sixty subjects ranging 14-17 years old were enrolled (obese: n = 30-50% females, and normal weight: n = 30-50% females). Stimulated saliva was collected for denaturing gradient gel electrophoresis (DGGE) band patterns and massive 16S rRNA gene sequencing using the Ion Torrent platform. Overall, data analysis revealed that male subjects harbored a higher diverse salivary microbiome, defined by a significant higher richness (32.48 versus 26.74) and diversity (3.36 versus 3.20), higher Simpson values (0.96 versus 0.95) and distinct bacterial community structure considering either sex or condition (p < 0.05). Bacterial community fingerprinting analysis in human saliva showed a positive correlation with increased body mass index (BMI) in adolescents. Veillonella, Haemophilus and Prevotella occurrence was found to be affected by BMI, whereas Neisseria and Rothia occurrence was significantly impacted by sex in obese subjects. Our findings suggest that male and female adolescents may harbor a naturally distinct salivary microbiota and that obesity may specifically have an impact on their oral bacterial community. The potential dysbiotic oral microbiome in obese adolescents raises new insights on the etiology and prevention of future conditions in these populations.

Leucine-Rich Amelogenin Peptide (LRAP) Uptake by Cementoblast Requires Flotillin-1 Mediated Endocytosis.

Basic, pre-clinical, and clinical studies have documented the potential of amelogenin, and its variants, to affect cell response and tissue regeneration. However, the mechanisms are unclear. Thus, the aim of the present study was to identify, in cementoblasts, novel binding partners for an alternatively spliced amelogenin form (Leucine-Rich Amelogenin Peptide-LRAP), which is supposed to act as a signaling molecule in epithelial-mesenchymal interactions. LRAP-binding protein complexes from immortalized murine cementoblasts (OCCM-30) were achieved by capture affinity assay (GST pull down) and proteins present in these complexes were identified by mass spectrometry and immunoblotting. Flotillin-1, which functions as a platform for signal transduction, vesicle trafficking, endocytosis, and exocytosis, was identified and confirmed by co-precipitation and co-localization assays as a protein-binding partner for LRAP in OCCM-30 cells. In addition, we found that exogenously added GST-LRAP recombinant protein was internalized by OCCM-30 cells, predominantly localized in the perinuclear region and, that inhibition of flotillin1-dependent functions by small interference RNA (siRNA) methodology significantly affected LRAP uptake and its biological properties on OCCM-30 cells, including LRAP effect on the expression of genes encoding osteocalcin (Ocn), bone sialoprotein (Bsp), and runt-related transcription factor 2 (RunX2). In conclusion, LRAP uptake by cementoblast involves flotillin-assisted endocytosis, which suggests an involvement of LRAP in lipid-raft-dependent signaling pathways which are mediated by flotillin-1. J. Cell. Physiol. 232: 556-565, 2017. © 2016 Wiley Periodicals, Inc.

Impact of the intermediary layer on sealant retention: a randomized 24-month clinical trial.

OBJECTIVES: The aims of this study were to assess long-term impact of tooth eruption stages (ES) on sealant retention on occlusal surfaces previously coated with intermediary bonding layer and to determine caries prevention. MATERIALS AND METHODS: Sixty-five school children were selected (aged 6-10 years), with four non-carious permanent first molar in different ES (OP (operculum present), ME (marginal edge), CE (completely erupted)). Split-mouth and single-blind study design was used. The teeth (260) were randomly selected according to treatment (sealant/technique): F (Fluroshield), H (Helioseal Clear Chroma), SF (Single Bond + F), EH (Excite + H). Sealant retention, marginal integrity, discoloration, and caries prevention were assessed after 6, 12, 18, and 24 months by calibrated examiner (Spearman = 0.91) using visual inspection. Data were submitted to the Cox proportional hazard model (survival analysis) and Likelihood ratio χ 2 test (correlation), p ≤ 0.05. RESULTS: At baseline, ES was 20 % in OP, 54 % in ME, and 26 % in CE. There was no significant difference on sealant retention between the treatments (p = 0.2774). However, significant differences were found regarding the ES on sealant retention (p = 0.0041). The CE stage showed the highest retention survival rate during the 24 months. The overall sealant prevention average was found to be about 99.4 % and showed no difference between the groups. CONCLUSIONS: Eruption stages affect sealant retention irregardless of the intermediate layer and type of sealant. However, there was caries prevention on tooth occlusal surfaces after 24 months, regardless of treatment. CLINICAL RELEVANCE: Sealing is recommended to prevent occlusal caries of newly erupted teeth in high-caries-risk patients. However, its application is critical on moisture limited control surfaces independently of sealer material and technique.

Titanium dioxide nanotubes applied to conventional glass ionomer cement influence the expression of immunoinflammatory markers: An in vitro study.

Objectives: To assess the impact of different concentrations TiO2-nt incorporated into a glass ionomer cement on the proliferation, mitochondrial metabolism, morphology, and pro- and anti-inflammatory cytokine production of cultured fibroblasts (NIH/3T3), whether or not stimulated by lipopolysaccharides (LPS-2 µg/mL, 24 h). Methods: TiO2-nt was added to KM (Ketac Molar EasyMix™, 3 %, 5 %, 7 % in weight); unblended KM was used as the control. The analyses included: Cell proliferation assay (n = 6; 24/48/72h); Mitochondrial metabolism assay (n = 6; 24/48/72h); Confocal laser microscopy (n = 3; 24/48/72h); Determination of biomarkers (IL-1ß/IL-6/IL-10/VEGF/TNF) by using both multiplex technology (n = 6; 12/18 h) and the quantitative real-time PCR assay (q-PCR) (n = 3, 24/72/120 h). The data underwent analysis using both the Shapiro-Wilk and Levene tests, and by generalized linear models (α = 0.05). Results: It demonstrated that cell proliferation increased over time, regardless of the presence of TiO2-nt or LPS, and displayed a significant increase at 72 h; mitochondrial metabolism increased (p < 0.05), irrespective of exposure to LPS (p = 0.937); no cell morphology changes were observed; TiO2-nt reverted the impact of KM on the secreted levels of the evaluated proteins and the gene expressions in the presence of LPS (p < 0.0001). Conclusions: TiO2-nt did not adversely affect the biological behavior of fibroblastic cells cultured on GIC discs.

Hybrid light applied with 37% carbamide peroxide bleaching agent with or without titanium dioxide potentializes color change effectiveness.

BACKGROUND: The effectiveness of dental color change was assessed by incorporating titanium dioxide (TiO2) into 37% carbamide peroxide bleaching agent associated with hybrid light. METHODOLOGY: Fifty bovine incisors were selected to receive the bleaching treatment, and separated into five groups (n = 10): 35% hydrogen peroxide (HP) (Whiteness HP, FGM/HP); 37% carbamide peroxide (CP) (Whiteness SuperEndo, FGM/CP); CP + hybrid light (HL) (CP HL); CP + 1% TiO2 (CP TiO2); CP TiO2 + hybrid light (CP TiO2 HL). The bleaching gels were applied to the dental surface for 30 min. Hybrid light (Whitening Plus, DMC/infrared laser diodes + blue LEDs +violet LEDs) was applied with 1 min of active light, alternating with 1 min of pause. A spectrophotometer (VITA Easyshade® Advance, Vita) was used to determine the color of the dental elements at baseline and time points after the 1st, 2nd and 3rd bleaching sessions. Color change effectiveness was evaluated using Vita Classical, CIEL*a*b*, WID and ΔEab, ΔE00 and ΔWID parameters. RESULTS: Generalized mixed linear models for repeated measures (α = 5%) showed significant decrease in Vita Classical scores and a* and b* values, as well as an increase in L* and ∆WID values for all the groups. Higher color change values for ΔEab were observed for CP HL and CP TiO2 HL, while those of ΔE00 and ΔWID were higher for CP TiO2 HL at the end of the bleaching treatment. CONCLUSION: Hybrid light applied with TiO2 incorporated into CP potentiated the effectiveness of the color change in the tooth structure.

Physicochemical evaluation of hydrogen peroxide bleaching gels containing titanium dioxide catalytic agent, and their influence on dental color change associated with violet LED.

BACKGROUND: The purpose of this study was: 1) to analyze the physical-chemical properties of hydrogen peroxide (HP) agents at 7.5% (HP7) and 35% (HP35), and the association with or without TiO2 nanotubes; 2) to evaluate dental bleaching effectiveness by using HP7 and HP35 together with or without TiO2 nanotubes, and applied with or without violet LED (VL). METHODOLOGY: 80 bovine incisors were treated according to groups (n = 10): HP35; HP35 + VL; HP35T (HP35 + TiO2); HP35T + VL; HP7; HP7 + VL; HP7T (HP7 + TiO2); HP7T + VL. Bleaching effectiveness was measured at 4 time points according to the Vita Classical, CIEL*a*b*, CIEDE2000, and WID parameters. HP35, HP35T, HP7, and HP7T were evaluated for mass change, pH, mean particle size (P), polydispersity (PDI), and zeta potential (ZP), over 6 months of storage. RESULTS: The pH of HP35 thickener was higher when associated to TiO2. At baseline, both of the bleaching gels containing TiO2 had lower P, PDI, and PZ (p < 0.05). All groups showed a significant decrease in Vita Classical color scores (p = 0.0037). There was a higher L* value, and lower b* values for HP7 when associated to VL after the 3rd session. (p < 0.05). HP35T showed higher color change (ΔEab, ΔE00), and lower a* value in the presence of VL (p < 0.05). ΔWID presented lower values for both gels, when TiO2 was incorporated (p ≤ 0.05). CONCLUSION: The incorporation of TiO2 to the bleaching gel showed good stability with minimal variations in physical-chemical properties. The color change in HP35 was more effective than in HP7, but the VL boosted the bleaching effectiveness of HP7, whereas TiO2 did not increase bleaching effectiveness.

TiO2 nanotube-containing glass ionomer cements display reduced aluminum release rates.

Titanium dioxide nanotubes (TiO2-nts) were incorporated into a glass ionomer cement (GIC) with improved mechanical properties and antibacterial activity. The aims of the present in vitro study were to define the elemental characterization, aluminum (Al) release rate, and initial working time for GIC reinforced with TiO2-nts, in an experimental caries model. TiO2-nts were incorporated into GIC powder components at 5% by weight, and compared with unblended GIC. Experimental approaches used energy-dispersive spectrometry (EDS), atomic absorption spectrophotometry (AAS), and brightness loss to define surface element properties, Al release rates, and initial working time, respectively. Statistical analysis was performed by 2-way ANOVA, Tukey's test, generalized linear models, and Student's t test (a = 0.05). EDS data analysis revealed that TiO2-nts incorporated into GIC had no significant impact on the typical elemental composition of GICs in an in vitro caries model. Regarding the demineralizing solution, GIC with TiO2-nt significantly decreased the Al release rate, compared with the control group (p < 0.0001). Moreover, TiO2-nt incorporated into GIC did not alter the initial working time of the material (p > 0.05). These findings add information to our scientific body of knowledge concerning the potential impact of TiO2-nt on the performance of conventional GICs.

Influence of different enamel substrates on microtensile bond strength of sealants after cariogenic challenge.

PURPOSE: To evaluate the microtensile bond strength (µTBS) of resin sealer on enamel substrates after cariogenic challenge. MATERIALS AND METHODS: Enamel blocks were obtained from human third molars and randomly divided into 6 groups (n = 10) according to enamel substrates (S: sound, CL: caries-like lesion, or CLTF: caries-like lesion + topical fluoride application) and sealant material (F: FluroShield, or H: Helioseal Clear Chroma). Sealants were placed on enamel surfaces, stored in 100% humidity (24 h, 37°C), and longitudinally sectioned into hourglass shapes. According to the groups, pH cycling was applied and the µTBS test was performed. The fracture patterns were assessed by SEM. RESULTS: Regarding substrates, the highest µTBS values in MPa were observed for CLTF enamel (26.0 ± 7.6), followed by S (22.0 ± 7.4) and CL (15.5 ± 4.9). A significant interaction was found between material and pH cycling (p = 0.0395). F (23.9 ± 7.6) showed higher µTBS values than H (18.3 ± 7.5) when submitted to pH cycling. The majority of samples presented mixed failure. CONCLUSIONS: Enamel substrate significantly affected µTBS, with the highest values for remineralized caries-like enamel lesions. Furthermore, µTBS values were dependent on both materials and pH cycling.

Fluoride release and remineralizing potential of varnishes in early caries lesions in primary teeth.

This study evaluated the fluoride (F) release and remineralizing potential of varnishes containing sodium fluoride (5% NaF), 5% NaF with CPP-ACP and 5% NaF with TCP in early caries lesions in primary teeth. To determine the F release at 1, 4, 6, 24, 72, and 168 hr, strips were covered with the varnishes and immersed in purified water (n = 7). The varnishes and purified water (negative control) were applied on enamel blocks with early caries lesions (n = 16). Enamel blocks were stored in artificial saliva and submitted to a pH-cycling. The area of enamel hardness loss (ΔS) was analyzed by microhardness, lesion depth by polarized light microscopy (PLM) and the chemical analysis by Energy-dispersive X-ray spectroscopy. Data were submitted to Shapiro-Wilk, two-way and one-way ANOVA, Tukey and paired t-tests (α = 5%). All varnishes released F, but 5% NaF with CPP-ACP had the highest release at 4, 6, 24, and 72 hr (p < .05) followed by 5% NaF with TCP and 5% NaF. No significant difference in ΔS was observed among varnishes (5% NaF = 4,098.4 ± 1,407.9; 5% NaF with CPP-ACP = 4,164.0 ± 1,019.3; 5% NaF with TCP = 4,183.2 ± 1,527.2; p = .999), but all of them differed from the negative control group (6,757.8 ± 2,274.7; p < .001). Lesion depth was lower in varnishes groups compared to negative control (% reduction: 5% NaF = 41.8%, 5% NaF with CPP-ACP = 38.8%, and 5% NaF with TCP = 36.3%; p < .001). Similar Ca, P, and Ca/P ratio percentages among groups and F was not detected after the treatments. All fluoride varnishes showed potential to enhance remineralization of early caries lesions in primary teeth.

Titanium dioxide nanotubes added to glass ionomer cements affect S. mutans viability and mechanisms of virulence.

This in vitro study evaluated the impact of TiO2 nanotubes (n-TiO2) incorporated into glass ionomer cement (GIC) on Streptococcus mutans (S. mutans) characteristics at cellular and molecular levels. n-TiO2, synthesized by the alkaline method (20 nm in size), was added to Ketac Molar EasyMix® at 0%, 3%, 5%, and 7% by weight. S. mutans strains were cultured on GIC disks with addition or not of n-TiO2 for 1, 3, and 7 days and the following parameters were assessed: inhibition halo (mm) (n=3/group); cell viability (live/dead) (n=5/group); cell morphology (SEM) (n=3/group); and gene expression by real-time PCR (vicR, covR, gtfB, gtfC, and gtfD) (n=6/group). The data were analyzed by the Kruskal-Wallis test, repeated-measures ANOVA or two-way ANOVA, and Tukey's and Dunn's post-hoc tests (α=0.05). The agar diffusion test showed a higher antibacterial property for 5% n-TiO2 compared with 3% and 7% (p<0.05) with no effect of time (1, 3, and 7 days). The cell number was significantly affected by all n-TiO2 groups, while viability was mostly affected by 3% and 5% n-TiO2, which also affected cell morphology and organization. Real-time PCR demonstrated that n-TiO2 reduced the expression of covR when compared with GIC with no n-TiO2 (p<0.05), with no effect of time, except for 3% n-TiO2 on vicR expression. Within-group and between-group analyses revealed n-TiO2 did not affect mRNA levels of gtfB, gtfC, and gtfD (p>0.05). Incorporation of n-TiO2 at 3% and 5% potentially affected S. mutans viability and the expression of key genes for bacterial survival and growth, improving the anticariogenic properties of GIC.

Review of the effects of infiltrants and sealers on non-cavitated enamel lesions.

PURPOSE: The aim of the present study was to perform a review of the effects of infiltrants and sealers on the inhibition of enamel demineralisation. MATERIALS AND METHODS: The authors searched the Cochrane Library, Embase, PubMed and Web of Science (ISI) for papers published between January 1970 and September 2008. The main search terms were 'artificial caries' or 'caries treatment' or 'caries-like lesion' or 'white spot lesion' or 'enamel demineralisation' or 'natural caries' and 'enamel' and 'sealant' or 'resin infiltration'. The inclusion criteria were studies that produced artificial non-cavitated enamel lesions before the application of sealant in in vivo or in vitro studies. Studies excluded were those that had not produced artificial non-cavitated enamel lesions before the application of sealant; had evaluated the inhibition of enamel demineralisation around restorations, sealant and orthodontic bracket/bands; had not evaluated the inhibition of enamel demineralisation after the sealant application; and had not applied sealant materials. Selected papers were given scores, from A to C, according to predetermined criteria. RESULTS: Eighteen studies were identified and included in the project critical appraisal. Two papers were classified as Grade A, nine as Grade B and seven as Grade C. CONCLUSIONS: It can be concluded that while fissure sealing acts as a diffusion barrier on the top of the lesion surface, the infiltration technique creates a barrier inside the lesion by replacing the mineral lost with a low-viscosity light-curing resin.

Titanium dioxide nanotubes incorporated into bleaching agents: physicochemical characterization and enamel color change.

OBJECTIVE: Titanium dioxide nanotubes are nanostructures that can accelerate the oxidation reaction of bleaching procedures and promote a more effective whitening effect. This study evaluated physicochemical properties of bleaching agents incorporated with titanium dioxide (TiO2) nanotubes, and the effects on tooth color change at different periods. METHODOLOGY: 40 premolars were treated according to the following groups (n=10): CP - 10% carbamide peroxide (1 hour daily/21 days); CPN - CP incorporated into TiO2; HP - 40% hydrogen peroxide (three 40-minute sessions/7 days apart); HPN - HP incorporated into TiO2. Color shade was evaluated at five different periods (baseline, after 7, 14 and 21 days of bleaching, and 7 days after end of treatment) according to Vita Classical, CIELab and CIEDE2000 scales. Mean particle size (P), polydispersity (PO) and zeta potential (ZP) were evaluated using dynamic light scattering. Data on the different variables were analyzed by mixed model tests for measures repeated in time (ZP e L*), generalized linear models for measures repeated in time (P, PO, Vita Classical and b*), and Friedman and Mann-Whitney tests (a* and color change/ΔE and ΔE00). RESULTS: CP and CPN presented higher P, higher PO and lower ZP than HP and HPN (p≤0.05). All groups showed a significant decrease in Vita Classical color scores after 7 days of bleaching (p<0.05), and HPN presented a greater significant reduction than the other groups. L* increased in TiO2 presence, in all groups, without any differences (p>0.05) in bleaching time. A significant reduction occurred in the a* and b* values for all the groups, and HPN presented lower a* and b* values (p<0.05) than CPN. ΔE was clinically noticeable after 7 days, in all groups, and all groups resulted in a perceptible color change according to ΔE00. CONCLUSION: TiO2 did not influence physicochemical properties of the bleaching agents. HPN presented more effective tooth bleaching than CPN.

TiO2 nanotubes improve physico-mechanical properties of glass ionomer cement.

OBJECTIVES: The aim of this study was to determine the physico-mechanical properties of a high viscosity glass ionomer cement (GIC) reinforced with TiO2 nanotubes (TiO2-nt). METHODS: TiO2-nt was incorporated into the GIC powder components (Ketac Molar EasyMix™) in concentrations of 0% (control group), 3%, 5%, 7% by weight. Compressive strength (n = 10/group), three point bending for flexural strength (n = 18/group), microshear bond strength to dentin and failure mode (n = 20/group), and surface roughness and weight loss before and after brushing simulation (30,000 cycles) (n = 8/group) were evaluated. Data were submitted to Shapiro-Wilk, ANOVA, Tukey and Chi-square tests (α ≤ 0.05). RESULTS: Addition of 5% of TiO2-nt into GIC presented the highest values for compressive strength and differed from the control, 3% and 7% groups (p = 0.023). There were no significant differences in flexural strength (p = 0.107) and surface roughness before and after the dental brushing (p = 0.287) among the groups. GIC added with 5% TiO2-nt showed the lowest weight loss values (p = 0.01), whereas the control, 3% or 5% TiO2-nt groups presented similar microshear bond strength values (p ≥ 0.05). The 5% TiO2-nt group featured higher microshear bond strength than the 7% TiO2-nt group (p = 0.034). Cohesive in material was the most representative failure mode for all groups. SIGNIFICANCE: The incorporation of TiO2-nt did not affect GIC's adhesiveness to dentin, but improved its compressive strength at 5%. Furthermore, TiO2-nt decreased the percentage of weight loss after GIC's surface wear.

Remineralizing effect of commercial fluoride varnishes on artificial enamel lesions.

The aim of this study was to evaluate soluble and insoluble fluoride concentrations in commercial varnishes, and their remineralization effect on artificial caries enamel lesions using surface and cross-sectional microhardness evaluations. Forty bovine enamel blocks were separated into four groups (n=10): control (no treatment), Enamelast (Ultradent Products), Duraphat (Colgate-Palmolive) and Clinpro White Varnish (3M ESPE). Surface enamel microhardness evaluations were obtained, artificial enamel caries lesions were developed by dynamic pH-cycling, and the varnishes were then applied every 6 days, after which the enamel blocks were submitted to dynamic remineralization by pH cycles. After removal of the varnishes, the enamel surfaces were reassessed for microhardness. The blocks were sectioned longitudinally, and cross-sectional microhardness measurements were performed at different surface depths (up to 300 µm depth). Polarized light microscopy images (PLMI) were made to analyze subsurface caries lesions. The fluoride concentration in whole (soluble and insoluble fluoride) and centrifuged (soluble fluoride) varnishes was determined using an extraction method with acetone. The data were analyzed to evaluate the surface microhardness, making adjustments for generalized linear models. There was a significant decrease in enamel surface microhardness after performing all the treatments (p<0.0001). Enamelast and Duraphat showed significantly higher enamel microhardness values than the control and the Clinpro groups (p = 0.0002). Microhardness loss percentage was significantly lower for Enamelast (p = 0.071; One-way ANOVA). PLMI showed that subsurface caries lesions were not remineralized with the varnish treatments. No significant differences in the in-depth microhardness levels (p = 0.7536; ANOVA) were observed among the treatments. Enamelast presented higher soluble and insoluble fluoride concentrations than the other varnishes (p < 0.0001; Kruskal-Wallis and Dunn tests). Enamelast and Duraphat varnishes promoted enamel surface remineralization, but no varnish remineralized the subsurface lesion body. Although insoluble and soluble fluoride concentration values did not correspond to those declared by the manufacturer, Enamelast presented higher fluoride concentration than the others.

A novel combination of biallelic ALPL mutations associated with adult hypophosphatasia: A phenotype-genotype association and computational analysis study.

Hypophosphatasia (HPP) is an inherited metabolic disorder that causes defective skeletal and dental mineralization. HPP exhibits a markedly heterogeneous range of clinical manifestations caused by dysfunction of the tissue-nonspecific isozyme of alkaline phosphatase (TNSALP), resulting from loss-of-function mutations in the ALPL gene. HPP has been associated with predominantly missense mutations in ALPL, and a number of compound heterozygous genotypes have been identified. Here, we describe a case of a subject with adult-onset HPP caused by a novel combination of missense mutations p.Gly473Ser and p.Ala487Val, resulting in chronic musculoskeletal pain, myopathy, persistent fatigue, vomiting, and an uncommon dental phenotype of short-rooted permanent teeth. Pedigree and biochemical analysis indicated that severity of symptoms was correlated with levels of residual ALP activity, and co-segregated with the p.Gly473Ser missense mutation. Bioinformatic analysis to predict the structural and functional impact of each of the point mutations in the TNSALP molecule, and its potential contribution to the clinical symptoms, revealed that the affected Gly473 residue is localized in the homodimer interface and predicted to have a dominant negative effect. The affected Ala487 residue was predicted to bind to Tyr479, which is closely located the N-terminal α-helix of TNSALP monomer 2, suggesting that both changes may impair dimer stability and catalytic functions. In conclusion, these findings assist in defining genotype-phenotype associations for HPP, and further define specific sites within the TNSALP molecule potentially related to neuromuscular manifestations in adult HPP, allowing for a better understanding of HPP pathophysiology.

Marginal adaptation of pit and fissure sealants after thermal and chemical stress. A SEM study.

PURPOSE: To evaluate the in vitro marginal adaptation (gap formation) in the fissure of different sealer materials (resin sealant, glass-ionomer cement, resin-modified glass-ionomer cement, and adhesive system) submitted to thermal and chemical stress, using scanning electron microscopy evaluation (SEM). METHODS: 80 impacted human third molars were randomly assigned to the following experimental groups (n = 10): FluroShield (F), Helioseal Clear Chroma (H), Vitremer (V), Fuji II-LC (FII), Ketac-Molar (KM), Fuji IX (FIX), Single Bond (SB), and Clearfil Protect Bond (CF). All groups were subjected to thermocycling and 14 days of pH cycling. A blinded and calibrated examiner performed SEM analysis. Gap formation was scored according to: 0 = no sealant marginal gaps; 1 = sealant marginal gaps present or total sealant loss. The score 0 was considered a success, while score 1 represented failure. Data were analyzed by Kruskal-Wallis and Bonferroni tests (P < 0.05). RESULTS: The success rates of SB (100%) and V (90%) were similar, but statistically superior to F (30%), H (20%), FIX (20%), and CF (0%) (P < 0.05). A tendency for similar behaviors of FII and KM to SB and V was observed and for similar behaviors of F, FIX and H to CF. The worst results were obtained for CF.

Effect of thermocycling on internal microhardness of high and low viscosity bulk fill composite resins in class I cavities / Efeito da termociclagem na microdureza interna de resinas compostas bulk fill de alta e baixa viscosidade aplicadas em cavidades classe I

ABSTRACT Objective: To evaluate the effect of thermocycling on the Knoop internal microhardness of high- and low viscosity bulk fill resins applied in Class I cavities. Methods: Thirty third molars with Class I cavity preparations were randomly divided into 3 groups according to the restorative system: nanoparticulated composite resin (Filtek™ Z350 XT 3M ESPE) (oblique increments of 2mm); low viscosity bulk fill resin (Filtek™ Bulk fill Flow, 3M ESPE) (3mm increments covered with a 1-mm layer of nanoparticulated resin); high viscosity bulk fill resin (Filtek™ Bulk fill, 3M ESPE) (single 4-mm increment). After 24h, half of samples was submitted to thermocycling (1,000 cycles). All samples (n=5) were sectioned in half to measure the internal microhardness at bottom and top of restoration. Results: Analysis of variance indicated that, for nanoparticulated composite resin, without thermocycling, the microhardness at the top was statistically lower than at the bottom. After thermocycling, there was no difference in microhardness between the top and the bottom. For low viscosity bulk fill resin, without thermocycling, there was no significant difference in microhardness means between the top and the bottom. After thermocycling, significantly higher microhardness was found at the top than at the bottom (p <0.05). For high viscosity bulk fill resin there was no significant difference between the microhardness values at the top and bottom, regardless of thermocycling (p> 0.05). In all composite resins, an increase in microhardness was observed after thermocycling (p <0.05). Conclusion: Thermocycling increased the internal microhardness of resin restorations, and, for the low viscosity bulk fill resin, the microhardness at the top was higher than at the bottom after thermocycling.

RESUMO Objetivo: Avaliar o efeito da termociclagem na microdureza Knoop interna de resinas bulk fill de alta e baixa viscosidade aplicadas em cavidades classe I. Métodos: Trinta terceiros molares com cavidades Classe I foram divididos aleatoriamente em três grupos de acordo com o sistema restaurador: Resina composta nanoparticulada (Filtek™ Z350 XT 3M ESPE) (incrementos oblíquos de 2mm); resina bulk fill de baixa viscosidade (Filtek™ Bulk fill Flow, 3M ESPE) (incremento de 3mm mais 1mm de resina nanoparticulada); resina bulk fill de alta viscosidade (Filtek ™ Bulk fill, 3M ESPE) (único incremento de 4mm). Após 24h, metade das amostras foi submetida a termociclagem (1.000 ciclos). Todas as amostras (n=5) foram seccionadas ao meio para mensuração da microdureza interna na base e topo da restauração. Resultados: Análise de variância indicou que, para a resina composta nanoparticulada e antes da termociclagem, a microdureza no topo foi estatisticamente inferior do que na base (p<0,05). Após a termociclagem, não houve diferença entre topo e base. Já para a resina bulk fill de baixa viscosidade, antes da termociclagem, não se constatou diferença significativa nos valores de microdureza entre topo e base. Após termociclagem, valores significativamente mais elevados de microdureza foram encontrados no topo do que na base (p<0,05). Para resina bulk fill de alta viscosidade, não houve diferença significativa na microdureza entre topo e base, realizando-se ou não termociclagem (p>0,05). Em todas as resinas compostas foi verificado aumento de microdureza após termociclagem (p<0,05). Conclusão: A termociclagem aumentou a microdureza para todas as resinas compostas, sendo que para a resina bulk fill de baixa viscosidade a microdureza no topo foi maior do que na base após termociclagem.