Self-assembled peptide P11-4 interacts with the type I collagen C-terminal telopeptide domain and calcium ions.

OBJECTIVES: Evaluate molecularly the role of P11-4 self-assembly peptide in dentin remineralization and its interaction with collagen I. METHODS: The calcium-responsive P11-4 peptide was analyzed by intrinsic fluorescence emission spectrum, circular dichroism spectrum (CD), and atomic force microscope (AFM). Differential light scattering was used to monitor the nucleation growth rate of calcium phosphate nanocrystals in the absence or in the presence of P11-4. AFM was used to analyze the radial size (nm) of calcium phosphate nanocrystals formed in the absence or in the presence of P11-4, as well as to verify the spatial structure of P11-4 in the absence or in the presence of Ca2+. RESULTS: The interaction of Ca2+ with the P11-4 (KD = 0.58 ± 0.06 mM) promotes the formation of ß-sheet antiparallel structure, leads to its precipitation in saturated solutions of Ca/P = 1.67 and induces the formation of parallel large fibrils (0.6 - 1.5 µm). P11-4 organized the HAP nucleation by reducing both the growth rate and size variability of nanocrystals, analyzed by the F test (p < 0.0001, N = 30). P11-4 interacts (KD = 0.75 ± 0.06 µM) with the KGHRGFSGL motif present at the C-terminal collagen telopeptide domain. P11-4 also increased the amount of HAP and collagen in the MDPC-23 cells. SIGNIFICANCE: The presented data propose a mechanism that will help future clinical and/or basic research to better understand a molecule able to inhibit structural collagen loss and help the impaired tissue to remineralize.

Self-assembling peptide-laden electrospun scaffolds for guided mineralized tissue regeneration.

OBJECTIVES: Electrospun scaffolds are a versatile biomaterial platform to mimic fibrillar structure of native tissues extracellular matrix, and facilitate the incorporation of biomolecules for regenerative therapies. Self-assembling peptide P11-4 has emerged as a promising strategy to induce mineralization; however, P11-4 application has been mostly addressed for early caries lesions repair on dental enamel. Here, to investigate P11-4's efficacy on bone regeneration, polymeric electrospun scaffolds were developed, and then distinct concentrations of P11-4 were physically adsorbed on the scaffolds. METHODS: P11-4-laden and pristine (P11-4-free) electrospun scaffolds were immersed in simulated body fluid and mineral precipitation identified by SEM. Functional groups and crystalline phases were analyzed by FTIR and XRD, respectively. Cytocompatibility, mineralization, and gene expression assays were conducted using stem cells from human exfoliated deciduous teeth. To investigate P11-4-laden scaffolds potential to induce in vivo mineralization, an established rat calvaria critical-size defect model was used. RESULTS: We successfully synthesized nanofibrous (∼ 500 nm fiber diameter) scaffolds and observed that functionalization with P11-4 did not affect the fibers' diameter. SEM images indicated mineral precipitation, while FTIR and XRD confirmed apatite-like formation and crystallization for P11-4-laden scaffolds. In addition, P11-4-laden scaffolds were cytocompatible, highly stimulated cell-mediated mineral deposition, and upregulated the expression of mineralization-related genes compared to pristine scaffolds. P11-4-laden scaffolds led to enhanced in vivo bone regeneration after 8 weeks compared to pristine PCL. SIGNIFICANCE: Electrospun scaffolds functionalized with P11-4 are a promising strategy for inducing mineralized tissues regeneration in the craniomaxillofacial complex.

Effect of a Self-Assembly Peptide on Surface Roughness and Hardness of Bleached Enamel.

After bleaching, enamel surfaces are damaged, contributing to erosion and tooth sensitivity. Although fluoride is used after bleaching to try and revert alterations, it is not capable of repairing tooth structure. This study compared the effect of a self-assembly peptide (P11-4), with and without fluoride, and sodium fluoride (NaF 2%) on the Knoop microhardness (KHN) and surface roughness (Ra (µm)) of bleached enamel with an in-office bleaching regimen. Enamel blocks of bovine teeth (5 × 5 × 2 mm) with standardized surface hardness were bleached with 35% carbamide peroxide, following the manufacturer's instructions. The teeth were randomly divided into the following groups (n = 7) according to post-bleaching treatment: no treatment (negative control) (C-); 2% NaF (NaF); Curodont™ Repair (Repair); and Curodont™ Protect (Protect). Specimens were stored in artificial saliva at 37 °C. To evaluate the effect of the post-bleaching treatments, KHN and Ra were measured before bleaching (baseline) and 24 h and 7 days after bleaching. Data were submitted to repeated measures ANOVA and Bonferroni tests (α = 0.05). There were significant interactions between the study factors (p = 0.001). After 7 days, Repair (572.50 ± 79.04) and Protect (583.00 ± 74.76) specimens showed increased surface KHN, with values higher than the NaF (465.50 ± 41.50) and C- (475.22 ± 58.95) baseline values. There was no significant difference in KHN at 24 h among groups (p = 0.587). At 24 h after bleaching, Repair was significantly different from all groups (p < 0.05). Repair showed the lowest Ra (µm) values (0.133 ± 0.035). After seven days, there was no significant difference in Ra values among groups when compared to the baseline. The use of P11-4-based materials after bleaching resulted in the fastest recovery to baseline enamel properties.

Comparative proteomic analysis of dental cementum from deciduous and permanent teeth.

BACKGROUND AND OBJECTIVES: Dental cementum (DC) is a mineralized tissue covering tooth roots that plays a critical role in dental attachment. Differences in deciduous vs. permanent tooth DC have not been explored. We hypothesized that proteomic analysis of DC matrix would identify compositional differences in deciduous (DecDC) vs. permanent (PermDC) cementum that might reflect physiological or pathological differences, such as root resorption that is physiological in deciduous teeth but can be pathological in the permanent dentition. METHODS: Protein extracts from deciduous (n = 25) and permanent (n = 12) teeth were pooled (five pools of DecDC, five teeth each; four pools of PermDC, three teeth each). Samples were denatured, and proteins were extracted, reduced, alkylated, digested, and analyzed by liquid chromatography-mass spectrometry (LC-MS/MS). The beta-binomial statistical test was applied to normalized spectrum counts with 5% significance level to determine differentially expressed proteins. Immunohistochemistry was used to validate selected proteins. RESULTS: A total of 510 proteins were identified: 123 (24.1%) exclusive to DecDC; 128 (25.1%) exclusive to PermDC; 259 (50.8%) commonly expressed in both DecDC and PermDC. Out of 60 differentially expressed proteins, 17 (28.3%) were detected in DecDC, including myeloperoxidase (MPO), whereas 43 (71.7%) were detected in PermDC, including decorin (DCN) and osteocalcin (BGLAP). Overall, Gene Ontology (GO) analysis indicated that all expressed proteins were related to GO biological processes that included localization and response to stress, and the GO molecular function of differentially expressed proteins was enriched in cell adhesion, molecular binding, cytoskeletal protein binding, structural molecular activity, and macromolecular complex binding. Immunohistochemistry confirmed the trends for selected differentially expressed proteins in human teeth. CONCLUSIONS: Clear differences were found between the proteomes of DecDC and PermDC. These findings may lead to new insights into developmental differences between DecDC and PermDC, as well as to a better understanding of physiological/pathological events such as root resorption.

Glass-ionomer cement modifies the gene expression of Streptococcus mutans providing a lower virulent biofilm.

PURPOSE: To evaluate the effect of glass-ionomer cement (GIC) on gene expression (gtfC, gtfD, covR, and vicR) of Streptococcus mutans (S. mutans) biofilms at 2, 4 and 24 hours. METHODS: Six groups were tested according to the materials and time observation, as follows: ceramic (IPS Empress Esthetic), as the control group, and GIC (Ketac Molar Easymix); and time points of S. mutans biofilm formation (2, 4, and 24 hours). Round-shaped samples (10 x 2 mm) of each material were prepared according to the manufacturers' specifications. GIC discs were handled in a laminar flow hood under aseptic conditions and stored at 100% relative humidity at 37°C for 24 hours to complete setting reaction. The samples were placed in a 24-well plate and immersed in 1.5 ml BHI + 1% sucrose with an inoculum of S. mutans UA159 to allow biofilm growth during 2, 4, and 24 hours. Next, the samples were removed, vortexed and centrifuged to collect cell pellets (n=5) for each material and time point. Pellets were stored at -80°C. Then, RNA was purified using the RNeasy Mini Kit protocol. The RNA was converted in cDNA using iScript cDNA Synthesis according to the manufacturer's recommendations. Analysis of gtfC, gtfD, vicR, and covR expressions was performed using Step One Real-Time qPCR device with specific primers for each gene and the analysis normalized by 16S reference gene expression. Data from gtfC, gtfD, and vicR were analyzed by t-test to compare between groups while Mann-Whitney was used to analyze covR expression (α= 0.05). RESULTS: No significant differences at 2 and 4 hours between materials for all analyzed genes were noted. However, in the 24-hour period, a significant decrease in gtfC and vicR expressions were observed, while covR expression increased when GIC was compared to ceramic. CLINICAL SIGNIFICANCE: The use of glass-ionomer cement decreased the virulence of S. mutans biofilms, which may imply a reduced bacterial cariogenic potential.

Wettability and pre-osteoblastic behavior evaluations of a dense bovine hydroxyapatite ceramics.

In this study, the wettability, cell viability, and roughness of an experimental dense bovine hydroxyapatite [Ca10(PO4)6(OH)2] ceramic block were evaluated so that, in the future, it could be used as a base material for dental implants. The results to commercial zirconia and a commercially pure titanium (Ti) alloy were compared. The surface roughness and contact angles were measured. An in vitro evaluation was conducted by means of tests in which pre-osteoblastic MC3T3-E1 cells were placed in indirect and direct contact with these materials. For cell viability, a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay and crystal violet test were conducted. A qualitative analysis was conducted using variable pressure scanning electron microscopy (SEM). No statistically significant differences were observed in wettability and roughness tests among the groups. In both the MTT assay and crystal violet test, all groups demonstrated satisfactory results without cytotoxicity. SEM showed cell adhesion and cell proliferation results on the material surfaces after 24 h and 48 h. In conclusion, this dense Ca10 (PO4)6(OH)2 ceramic can be considered as a potential biocompatible material.

Influence of the incorporation of triclosan methacrylate on the physical properties and antibacterial activity of resin composite.

The incorporation of antimicrobials in the composites as an attempt to reduce bacterial adhesion without jeopardizing mechanical properties is a challenge for Dentistry. OBJECTIVE To evaluate the bacterial adhesion and physical properties of a composite containing the methacrylate triclosan- derivative monomer (TM). METHODOLOGY TM was synthesized and added to an experimental composite. Samples were divided into two groups: Control and TM (13.4 wt%). Antibacterial Activity: Three specimens of each material were prepared and placed on bacterial suspensions of Streptococcus mutans for 1, 5 and 10 days. After these periods the counting of the colonies (log10) was performed. Assays was performed in triplicate. Physical Properties: Three-body Abrasion (TBA): Ten specimens of each material were prepared and stored at 37°C/24 h. The surface roughness (Ra) and hardness (KHN) were analyzed. Next, the specimens were submitted to abrasive wear (30,000 cycles) and re-evaluated for Ra and KHN; Sorption/solubility (SS): cylindrical specimens (n=10) were prepared and weighted. The specimens were immersed in deionized water for 7 days at 37°C and then their weight was verified again. SS were calculated using accepted formulas; Diametral tensile strength (DTS): specimens (n=10) underwent test performed in an Instron universal testing machine at a crosshead speed of 1 mm/min. Data were submitted to appropriate statistical tests according to data distribution and assay (p<0.05). RESULTS Bacterial Adhesion: TM showed a significant reduction on biofilm accumulation in the evaluated periods: 1 day (1.537±0.146); 5 days (2.183±0.138) and 10 days (4.469±0.155) when compared with Control: 1 day (4.954±0.249); 5 days (5.498±0.257) and 10 days (6.306±0.287). Physical Properties: For TBA, SS and DTS no significant difference was found between groups Control and TM. The incorporation of methacrylate triclosan-based monomer in the experimental composite reduce bacterial adhesion of S. mutans and did not affect important polymer properties.

Membrane proteome characterization of periodontal ligament cell sets from deciduous and permanent teeth.

BACKGROUND: Physiological roles for the periodontal ligament (PDL) include tooth eruption and anchorage, force absorption, and provision of proprioceptive information. Despite the advances in understanding the biology of PDL cells, there is a lack of information regarding the molecular signature of deciduous (DecPDL) and permanent (PermPDL) PDL tissues. Thus, the present study was designed to characterize the membrane proteome of DecPDL and PermPDL cells. METHODS: Primary PDL cells were obtained (n = 6) and a label-free quantitative proteome of cell membrane-enriched components was performed. Proteome findings were validated by quantitative polymerase chain reaction and Western blot assays in fresh human tissues (n = 8) and primary cell cultures (n = 6). In addition, confocal microscopy was used to verify the expression of target factors in the PDL cell cultures. RESULTS: Comparative gene ontology enrichment analysis evidenced that most stickling differences involved "endomembrane system" (PICALM, STX4, and LRP10), "hydrolase activity" (NCSTN and XRCC6), "protein binding" (PICALM, STX4, GPNMB, VASP, extended-synaptotagmin 2 [ESYT2], and leucine-rich repeat containing 15 [LRRC15]), and "isomerase activity" (FKBP8). Data are available via ProteomeXchange with identifier PXD010226. At the transcript level, high PICALM in DecPDL and ESYT2 and LRRC15 in PermPDL were confirmed in fresh PDL tissues. Furthermore, Western blot analysis confirmed increased levels of PICALM, LRRC15, and ESYT2 in cells and/or fresh tissues, and confocal microscopy confirmed the trends for PICALM and LRRC15 expression in PDL cells. CONCLUSION: We report the first comprehensive characterization of the membrane protein machinery of DecPDL and PermPDL cells, and together, we identified a distinct molecular signature for these cell populations, including unique proteins for DecPDL and PermPDL.

Synthesis and application of triclosan methacrylate monomer in resin composites.

OBJECTIVES: To evaluate the antibacterial activity, bacterial viability, cytotoxicity, and mechanical/physical properties of a novel methacrylate triclosan-derivative monomer (TM) incorporated in dental resin composite. METHODS: TM was synthesized by esterification and, after characterization by FT-IR, was added to an experimental composite. Samples were divided into two groups according to TM presence, i.e., C1 (control) and C2 (C1 + 14.4% TM). Microbiological properties: Specimens (C1 and C2) were prepared and placed on bacterial suspensions of Streptococcus mutans. Antibacterial activity, MTT, and live/dead bacterial viability were used to test the resin composites. All assays were performed in triplicates. Mechanical properties: Specimens underwent compression (CS) and flexural strength (FS) tests conducted in an Instron universal testing machine at a crosshead speed of 0.5 mm/min. Physical properties: Specimens were assessed for Knoop hardness (KHN) and crosslink density (CD). Fourier transform infrared spectroscopy allowed the degree of conversion (DC) to be evaluated. Data were subjected to appropriate statistical tests according to data distribution and assay (p < 0.05). RESULTS: Microbiological properties: C2 showed the lowest biofilm accumulation and the highest membrane-compromised bacteria in the biofilm. Mechanical/physical properties: For CS, FS, KHN, and DC, there was no significant difference between groups C1 and C2; however, significant difference was observed for the CD assay. CONCLUSIONS: The triclosan methacrylate reduces bacterial adhesion of S. mutans and decreased the formation of bacterial biofilm without affecting important polymer properties. The triclosan methacrylate incorporated in resin composite could greatly reduce the live bacterial adhesion of S. mutans and decrease the formation of bacterial biofilm without affecting important polymer properties. CLINICAL SIGNIFICANCE: The resin composites containing triclosan methacrylate could greatly reduce the bacterial adhesion and biofilm formation. That might prevent the secondary caries round the margins of the restorations.

Influence of the incorporation of triclosan methacrylate on the physical properties and antibacterial activity of resin composite

Abstract The incorporation of antimicrobials in the composites as an attempt to reduce bacterial adhesion without jeopardizing mechanical properties is a challenge for Dentistry. Objective: To evaluate the bacterial adhesion and physical properties of a composite containing the methacrylate triclosan- derivative monomer (TM). Methodology: TM was synthesized and added to an experimental composite. Samples were divided into two groups: Control and TM (13.4 wt%). Antibacterial Activity: Three specimens of each material were prepared and placed on bacterial suspensions of Streptococcus mutans for 1, 5 and 10 days. After these periods the counting of the colonies (log10) was performed. Assays was performed in triplicate. Physical Properties: Three-body Abrasion (TBA): Ten specimens of each material were prepared and stored at 37°C/24 h. The surface roughness (Ra) and hardness (KHN) were analyzed. Next, the specimens were submitted to abrasive wear (30,000 cycles) and re-evaluated for Ra and KHN; Sorption/solubility (SS): cylindrical specimens (n=10) were prepared and weighted. The specimens were immersed in deionized water for 7 days at 37°C and then their weight was verified again. SS were calculated using accepted formulas; Diametral tensile strength (DTS): specimens (n=10) underwent test performed in an Instron universal testing machine at a crosshead speed of 1 mm/min. Data were submitted to appropriate statistical tests according to data distribution and assay (p<0.05). Results: Bacterial Adhesion: TM showed a significant reduction on biofilm accumulation in the evaluated periods: 1 day (1.537±0.146); 5 days (2.183±0.138) and 10 days (4.469±0.155) when compared with Control: 1 day (4.954±0.249); 5 days (5.498±0.257) and 10 days (6.306±0.287). Physical Properties: For TBA, SS and DTS no significant difference was found between groups Control and TM. The incorporation of methacrylate triclosan-based monomer in the experimental composite reduce bacterial adhesion of S. mutans and did not affect important polymer properties.

In-vitro comparative study of the use of 2 % chlorhexidine on microtensile bond strength of different dentin adhesives: a 6 months evaluation / Estudio comparativo in vitro del uso de clorhexidina al 2 % sobre la resistencia de unión microtensil de diferentes adhesivos dentinarios: una evaluación de 6 meses

The aim of this study was to evaluate the effect of chlorhexidine (CHX) dentin treatment on microtensile bond strength (µTBS) of adhesive systems in different storage times. Occlusal enamel was removed from ninety third-molars and flat surfaces of middle dentin were exposed. Teeth were randomly divided in 6 groups according to adhesive system (etch-and-rinse : Adper Scotchbond 1XT - ASB ; self-etch: Adper Prompt L-Pop ­ APP; and universal: Single Bond Universal - SBU) and chlorhexidine (CHX) dentin treatment (2 % CHX application for 20 s prior Primer). After resin composite build up, teeth were sectioned to obtain beam specimens and divided in 3 subgroups (n=5): 72h, 3 and 6 months storage times. After the storage times, teeth were tested in tension until failure (0.5 mm/min). SEM was performed to observe hybrid layer of adhesive systems. Data were analyzed using one-way ANOVA and Tukey tests. At 72 h, all equivalent groups (same adhesive system, different dentin treatments) maintained their µTBS when compared CHX-treatment. At 3 and 6 months, non-treated CHX groups showed less µTBS than CHX-treated ones. Six months storage time did not significantly decrease µTBS, except for G2-ASB. The effect of CHX on dentin µTBS depends on storage times and adhesive systems. While immediate µTBS was not affected by CHX treatment, CHX improved dentin µTBS after 3 and 6 months.

El objetivo de este estudio fue evaluar el efecto del tratamiento con clorhexidina (CHX) de la dentina sobre la resistencia de la unión microtensil (mTBS) de los sistemas adhesivos en diferentes tiempos de almacenamiento. Se retiró el esmalte oclusal de noventa terceros molares y se expusieron superficies planas de la dentina media. Los dientes se dividieron al azar en 6 grupos de acuerdo con el sistema adhesivo (con grabado ácido: Adper Scotchbond 1XT-ASB, auto-grabado: Adper Prompt L-Pop-APP y universal: Single Bond Universal- SBU) y el tratamiento de la dentina con clorhexidina (CHX) (aplicación de CHX al 2 % 20 s antes del Primer). Después de la aplicación de la resina compuesta, los dientes fueron seccionados para obtener muestras en forma de barras y divididos en 3 subgrupos (n = 5) con tiempos de almacenamiento de 72 h, 3 y 6 meses. Después de los tiempos de almacenamiento, los dientes se sometieron a tensión hasta la fractura (0,5 mm / min). SEM se realizó para observar la capa híbrida de sistemas adhesivos. Los datos se analizaron mediante ANOVA unidireccional y pruebas de Tukey. A las 72 h, todos los grupos equivalentes (el mismo sistema adhesivo, diferentes tratamientos de dentina) mantuvieron su mTBS cuando se comparó el tratamiento CHX. A los 3 y 6 meses, los grupos CHX no tratados mostraron menos mTBS que los tratados con CHX. Seis meses de tiempo de almacenamiento no disminuyó significativamente el mTBS, excepto para el G2-ASB. El efecto de CHX sobre la dentina mTBS depende del tiempo de almacenamiento y de los sistemas adhesivos. Mientras que el mTBS inmediato no se vio afectado por el tratamiento con CHX, CHX mejoró la mTBS a dentina después de 3 y 6 meses.

Effect of 2 % chlorhexidine on dentin shear bond strength of different adhesive systems: a 6 months evaluation / Efecto de la clorhexidina al 2 % sobre la resistencia al cizallamiento de diferentes sistemas adhesivos: una evaluación de 6 meses

This study evaluated the effect of chlorhexidine dentin treatment on shear bond strength (SBS) of adhesive systems after different storages. The work included 144 third molars that had their dentin exposed and were divided in 6 groups: G1 (ASB+CHX: Adper Scotchbond 1XT + chlorhexidine 2 % prior Primer); G2(ASB); G3 (APP+CHX: Adper Prompt L-Pop + CHX); G4(APP); G5 (SBU+CHX: Single Bond Universal + CHX); and G6(SBU). Resin build-up was performed and teeth were subdivided regarding storage times (n=8): 72 h, 3 and 6 months. Next, SBS test was performed. At 72 hours, all equivalent groups (same adhesive system, different dentin treatment) showed no significant difference in SBS (P.05). Self-etch adhesive groups (with or without CHX) presented lower SBS compared to other systems (P.05). After 3 and 6 months, all CHX-treated groups presented significantly higher SBS compared to equivalent non-treated groups (P.05). For both storage times, Single Bond Universal presented the highest SBS values within the same dentin treatment (P.05), while Adper Scotchbond and Adper Prompt-L-Pop were not significantly different among them, also within the same dentin treatments [3 months (with CHX: P=.966; without: P=.958) and 6 months (with CHX: P =.887; without: P=.990)]. CHX Dentin disinfection is indicated for all classes of adhesives studied.

Este estudio evaluó el efecto del tratamiento de la dentina con clorhexidina sobre la resistencia al cizallamiento (SBS) de sistemas adhesivos después de diferentes almacenamientos. Se removió el esmalte oclusal a 144 terceros molares y se dejó su dentina media expuesta, posteriormente se dividieron al azar en 6 grupos: G1 (ASB + CHX: Adper Scotchbond 1XT + clorhexidina 2 % antes del Primer); G2 (ASB); G3 (APP + CHX: L-Pop + CHX de Adper); G4 (APP); G5 (SBU + CHX: Single Bond Universal + CHX); y G6 (SBU). Se realizó la aplicación de la resina compuesta y se subdividieron los grupos con respecto a los tiempos de almacenamiento (n = 8): 72h, 3 y 6 meses. A continuación, se realizó la prueba SBS. A las 72 horas, todos los grupos equivalentes (el mismo sistema adhesivo, diferentes tratamientos de dentina) no mostraron diferencias significativas en los valores de SBS (P.05). Los grupos de adhesivo de auto-grabado (con o sin CHX) presentaron valores de SBS más bajos en comparación con otros sistemas (P.05). Después de 3 y 6 meses, todos los grupos tratados con CHX presentaron valores de SBS significativamente mayores en comparación con los grupos no tratados equivalentes (P.05). Para ambos tiempos de almacenamiento, Single Bond Universal presentó los valores de SBS más altos dentro del mismo tratamiento dentinario (P.05), mientras que el Adper Scotchbond y el Adper Prompt-L-Pop no fueron significativamente diferentes entre ellos, también dentro de los mismos tratamientos dentinarios 3 meses (con CHX: P = .966, sin: P = .958) y 6 meses (con CHX: P = .887; sin: P = .990). La desinfección de la dentina con CHX está indicada para todas las clases de adhesivos estudiados.

Effect of blue and violet light on polymerization shrinkage vectors of a CQ/TPO-containing composite.

OBJECTIVE: To evaluate the effect of light-curing wavelengths on composite filler particle displacement, and thus to visualize localized polymerization shrinkage in a resin-based composite (RBC) containing camphorquinone (CQ) and Lucirin TPO (TPO). METHODS: Three light-curing units (LCUs) were used to light-cure a RBC containing CQ and TPO: a violet-only, a blue-only, and a dual-wavelength, conventional (Polywave®, emitting violet and blue wavelengths simultaneously). Zirconia fillers were added to the RBC to act as filler particle displacement tracers. LCUs were characterized for total emitted power (mW) and spectral irradiant output (mW/cm2/nm). 2-mm high, 7-mm diameter silanized glass cylindrical specimens were filled in a single increment with the RBC, and micro-computed tomography (µ-CT) scans were obtained before and after light-curing, according to each LCU (n=6). Filler particle movement identified polymerization shrinkage vectors, traced using software, at five depths (from 0 up to 2mm): top, top-middle, middle, middle-bottom and bottom. RESULTS: Considering different RBC depths within the same LCU, use of violet-only and conventional LCUs showed filler particle movement decreased with increased depth. Blue-only LCU showed homogeneous filler particle movement along the depths. Considering the effect of different LCUs within the same depth, filler particle movement within LCUs was not statistically different until the middle of the samples (P>.05). However, at the middle-bottom and bottom depths (1.5 and 2mm, respectively), blue-only LCU compared to violet-only LCU showed higher magnitude of displacement vector values (P<.05). Use of the conventional LCU showed filler displacement magnitudes that were not significantly different than blue-only and violet-only LCUs at any depth (P>.05). With respect to the direction of particle movement vectors, use of violet-only LCU showed a greater displacement when close to the incident violet LED; blue-only LCU showed equally distributed particle displacement values within entire depth among the samples; and the conventional LCU showed greater filler displacement closer to the blue LED locations. SIGNIFICANCE: Filler particle displacement in a RBC as a result of light-curing is related to localized application of light wavelength and total emitted power of the light emitted on the top surface of the RBC. When the violet LED is present (violet-only and conventional LCUs), filler particle displacement magnitude decreased with increased depth, while results using the blue-only LED show a more consistent pattern of displacement. Clinically, these results correlate to production of different characteristics of curing within a RBC restoration mass, depending on localized wavelengths applied to the irradiated surface.

Streptococcus Mutans Biofilm Influences on the Antimicrobial Properties of Glass Ionomer Cements.

The aim of this study was to evaluate the in vitro antibacterial and biofilm inhibition properties of glass ionomer restorative cements. Ketac Nano, Vitremer, Ketac Molar Easymix and Fuji IX were analyzed using the following tests: a) agar plate diffusion test to evaluate the inhibitory activity of cements against S. mutans (n=8); b) S. mutans adherence test by counting colony-forming units after 2 h of material/bacteria exposure (n=10); c) biofilm wet weight after seven days of bacterial accumulation on material disks, with growth medium renewed every 48 h (n=10); d) pH and fluoride measurements from the medium aspired at 48 h intervals during the 7-day biofilm development (n=10). Data from the a, b and c tests were submitted to Kruskal-Wallis and Mann-Whitney tests and the fluoride-release and pH data were submitted to two-way ANOVA and Tukey tests (a=5%). Vitremer followed by Ketac Nano showed the greatest inhibitory zone against S. mutans than the conventional ionomers. Vitremer also showed higher pH values than Ketac Nano and Fuji IX in the first 48 h and released higher fluoride amount than Ketac Nano e Ketac Molar Easymix throughout the experimental period. The chemical composition of restorative glass ionomer materials influenced the antibacterial properties. The resin modified glass ionomer (Vitremer) was more effective for inhibition of S. mutans and allowed greater neutralization of the pH in the first 48 h. However, the type of glass ionomer (resin modified or conventional) did not influence the weight and adherence of the biofilm and fluoride release.

Streptococcus Mutans Biofilm Influences on the Antimicrobial Properties of Glass Ionomer Cements

Abstract The aim of this study was to evaluate the in vitro antibacterial and biofilm inhibition properties of glass ionomer restorative cements. Ketac Nano, Vitremer, Ketac Molar Easymix and Fuji IX were analyzed using the following tests: a) agar plate diffusion test to evaluate the inhibitory activity of cements against S. mutans (n=8); b) S. mutans adherence test by counting colony-forming units after 2 h of material/bacteria exposure (n=10); c) biofilm wet weight after seven days of bacterial accumulation on material disks, with growth medium renewed every 48 h (n=10); d) pH and fluoride measurements from the medium aspired at 48 h intervals during the 7-day biofilm development (n=10). Data from the a, b and c tests were submitted to Kruskal-Wallis and Mann-Whitney tests and the fluoride-release and pH data were submitted to two-way ANOVA and Tukey tests (a=5%). Vitremer followed by Ketac Nano showed the greatest inhibitory zone against S. mutans than the conventional ionomers. Vitremer also showed higher pH values than Ketac Nano and Fuji IX in the first 48 h and released higher fluoride amount than Ketac Nano e Ketac Molar Easymix throughout the experimental period. The chemical composition of restorative glass ionomer materials influenced the antibacterial properties. The resin modified glass ionomer (Vitremer) was more effective for inhibition of S. mutans and allowed greater neutralization of the pH in the first 48 h. However, the type of glass ionomer (resin modified or conventional) did not influence the weight and adherence of the biofilm and fluoride release.

Resumo O objetivo neste estudo foi avaliar in vitro as propriedades antibacterianas e a inibição do biofilme de cimentos de ionômero de vidro restauradores. Ketac Nano, Vitremer, Ketac Molar Easymix and Fuji IX foram avaliados através dos seguintes testes: a) teste de difusão em ágar para avaliar a inibição de S. mutans nos cimentos (n=8); b) adesão de S. mutans pela contagem de unidades formadoras de colônia após 2h de exposição material/bactéria (n=10); c) peso do biofilme úmido após sete dias de acúmulo bacteriano nos discos do material, com meio de cultura renovado após 48 h (n=10); d) mensuração do pH e liberação de flúor do meio aspirado nos intervalos de 48 h durante 7 dias de crescimento do biofilme (n=10). Os dados dos testes a, b e c foram submetidos aos testes Kruskal-Wallis e Mann-Whitney e os dados de liberação de flúor e pH a ANOVA dois fatores e Tukey (a = 5%). Vitremer seguido pelo Ketac Nano mostrou maior zona de inibição contra S. mutans quando comparados aos ionômeros convencionais. Vitremer também apresentou valores de pH mais elevados do que Ketac Nano e Fuji IX nas primeiras 48 h e liberou maior quantidade de flúor do que Ketac Nano e Ketac Molar Easymix durante todo o período experimental. A composição química dos ionômeros de vidro restauradores influenciou nas propriedades antibacterianas. O ionômero de vidro modificado por resina (Vitremer) foi mais eficaz na inibição de S. mutans e permitiu maior neutralização do pH nas primeiras 48 h. No entanto, o tipo de ionômero de vidro (modificado por resina ou convencional) não influenciou no peso e adesão do biofilme e na liberação de flúor.

Secretome Profiling of Periodontal Ligament from Deciduous and Permanent Teeth Reveals a Distinct Expression Pattern of Laminin Chains.

It has been suggested that there are histological and functional distinctions between the periodontal ligament (PDL) of deciduous (DecPDL) and permanent (PermPDL) teeth. Thus, we hypothesized that DecPDL and PermPDL display differences in the constitutive expression of genes/proteins involved with PDL homeostasis. Primary PDL cell cultures were obtained for DecPDL (n = 3) and PermPDL (n = 3) to allow us to perform label-free quantitative secretome analysis. Although a highly similar profile was found between DecPDL and PermPDL cells, comparative secretome analysis evidenced that one of the most stickling differences involved cell adhesion molecules, including laminin subunit gamma 1 (LAMC1) and beta 2 (LAMB2). Next, total RNA and protein extracts were obtained from fresh PDL tissues of deciduous (n = 6) and permanent (n = 6) teeth, and Western blotting and qPCR analysis were used to validate our in vitro findings. Western blot analysis confirmed that LAMC1 was increased in DecPDL fresh tissues (p<0.05). Furthermore, qPCR data analysis revealed that mRNA levels for laminin subunit beta 1 (LAMB1), beta 3 (LAMB3), LAMC1, and gamma 2 (LAMC2) were higher in DecPDL fresh tissues, whereas transcripts for LAMB2 were increased in PermPDL (p<0.05). In conclusion, the differential expression of laminin chains in DecPDL and PermPDL suggests an involvement of laminin-dependent pathways in the control of physiological differences between them.

Comparative Analysis of Root Dentin Morphology and Structure of Human Versus Bovine Primary Teeth.

This study evaluated the structural and morphological differences between human and bovine primary root canals. Primary human maxillary central incisors (H) (n=9) and primary bovine incisors (B) (n=9) were selected. The roots were sectioned in the vestibular-lingual direction, planed and delimited in cervical, middle, and apical thirds. Tubule density (number of tubules per mm2) and diameter were analyzed by scanning electron microscopy (1,000 and 5,000×) using Image J 1.47 software. Data were submitted to two-way repeated measures ANOVA and Tukey tests (α=0.05). The highest tubule density was observed for B (28.527±1.717 mm2) compared with H (15.931±0.170 mm2) (p<0.01). Regarding root thirds, the cervical third presented a greater tubule density (26.417±11.654 mm2) than the apical third (17.999±5.873 mm2). The diameter of the dentin tubules was not different for cervical (3.50±0.08 µm), middle (3.45±0.30 µm) and apical thirds (3.42±0.33 µm) and substrate (H-3.29±0.14 µm; B-3.63±0.06 µm). It could be concluded that: (1) the radicular dentin structure of human and bovine primary teeth and root thirds differ in terms of the tubule density; (2) the radicular dentin morphology of human and bovine primary teeth and root thirds are similar in terms of the diameter of the dentin tubules.

In situ bacterial accumulation on dental restorative materials. CLSM/COMSTAT analysis.

PURPOSE: To evaluate the influence of different restorative materials on the biofilm structure accumulated in situ. METHODS: 15 discs of each material (ceramic; resin composite; resin-modified and conventional glass-ionomers; amalgam) were adapted to palatal devices in order to accumulate biofilm in situ, under a cariogenic challenge (20% sucrose solution, 10x/day). After 7 days, the specimens were carefully removed and visualized by confocal laser scanning microscopy (CLSM). The images were analyzed qualitatively (descriptive analysis about cell viability and architecture) and quantitatively using COMSTAT software (area, bio-volume, mean thickness, maximum thickness and roughness coefficient of the biofilm). The statistical analysis was performed by using the Kolmogorov-Smirnov and Kruskal-Wallis tests (P ≤ 5%). RESULTS: The medians of the biofilm parameters analyzed showed no statistical difference regarding different materials. However, qualitatively, glass-ionomer cements and amalgam showed visually a prevalence of non-viable cells forming small clusters distributed by the biofilm, and voids were presented in smaller proportion in the biofilm volume compared to composite and ceramic.

Inhibitory effects of a cured antibacterial bonding system on viability and metabolic activity of oral bacteria.

OBJECTIVES: To evaluate the antimicrobial efficacy of Clearfil SE Protect (CP) and Clearfil SE Bond (CB) after curing and rinsed against five individual oral microorganisms as well as a mixture of bacterial culture prepared from the selected test organisms. METHODS: Bacterial suspensions were prepared from single species of Streptococcus mutans, Streptococcus sobrinus, Streptococcus gordonii, Actinomyces viscosus and Lactobacillus lactis, as well as mixed bacterial suspensions from these organisms. Dentin bonding system discs (6 mm×2 mm) were prepared, cured, washed and placed on the bacterial suspension of single species or multispecies bacteria for 15, 30 and 60 min. MTT, Live/Dead bacterial viability (antibacterial effect), and XTT (metabolic activity) assays were used to test the two dentin system's antibacterial effect. All assays were done in triplicates and each experiment repeated at least three times. Data were submitted to ANOVA and Scheffe's f-test (5%). RESULTS: Greater than 40% bacteria killing was seen within 15 min, and the killing progressed with increasing time of incubation with CP discs. However, a longer (60 min) period of incubation was required by CP to achieve similar antimicrobial effect against mixed bacterial suspension. CB had no significant effect on the viability or metabolic activity of the test microorganisms when compared to the control bacterial culture. CP was significantly effective in reducing the viability and metabolic activity of the test organisms. SIGNIFICANCE: The results demonstrated the antimicrobial efficacy of CP both on single and multispecies bacterial culture. CP may be beneficial in reducing bacterial infections in cavity preparations in clinical dentistry.

Bonding orthodontics brackets to enamel using experimental composites with an iodonium salt.

OBJECTIVES: This study investigated whether the addition of diphenyliodonium chloride (DPC) to experimental resin bonding agents would allow fixation of brackets to enamel using shorter light exposure times. METHODS: Photoactivated dimethacrylate-based composites were prepared containing DPC molar concentrations of 0 (control), 0.5 (R05), or 1 per cent (R1). Metallic brackets were bonded to bovine incisors and the bond strengths were evaluated using a shear test. In total, 18 groups were tested (n = 15 per group) defined by three bonding materials (control, R05, or R1), three light-activation time (8, 20, or 40 seconds), and two storage periods (10 minutes or 24 hours). The adhesive remnant index (ARI) was scored under magnification. Data were statistically analysed at a 5 per cent significance level. RESULTS: At 10 minutes, the control composite showed lower bond strengths than the DPC-modified bonding agents for all light-activation times. Differences in bond strengths between both DPC-modified agents were not significant. Lower bond strengths at 10 minutes were generally observed for groups light activated for 8 seconds compared with groups light activated for 20 and 40 seconds. At 24 hours, no significant differences were observed among the light-activation times. The bond strengths at 24 hours were higher than the bond strengths at 10 minutes for all groups. A predominance of ARI scores 2 and 3 was generally observed. CONCLUSION: The use of a ternary photoactivation system containing an iodonium salt in bonding composites may allow bonding brackets to enamel using reduced light exposure times.