Laser Photobiomodulation 808 nm: Effects on Gene Expression in Inflammatory and Osteogenic Biomarkers in Human Dental Pulp Stem Cells.

The tissue engineering of dental oral tissue is tackling significant advances and the use of stem cells promises to boost the therapeutical approaches of regenerative dentistry. Despite advances in this field, the literature is still scarce regarding the modulatory effect of laser photobiomodulation (PBM) on genes related to inflammation and osteogenesis in Postnatal Human Dental Pulp Stem cells (DPSCs). This study pointedly investigated the effect of PBM treatment in proliferation, growth and differentiation factors, mineralization, and extracellular matrix remodeling genes in DPSCs. Freshly extracted human third molars were used as a source for DPSCs isolation. The isolated DPSCs were stimulated to an inflammatory state, using a lipopolysaccharide (LPS) model, and then subjected or not to laser PBM. Each experiment was statistically evaluated according to the sample distribution. A total of 85 genes related to inflammation and osteogenesis were evaluated regarding their expression by RT-PCR. Laser PBM therapy has shown to modulate several genes expression in DPSCs. PBM suppressed the expression of inflammatory gene TNF and RANKL and downregulated the gene expression for VDR and proteolytic enzymes cathepsin K, MMP-8 and MMP-9. Modulation of gene expression for proteinase-activated receptors (PARs) following PBM varied among different PARs. As expected, PBM blocked the odontoblastic differentiation of DPSCs when subjected to LPS model. Conversely, PBM has preserved the odontogenic potential of DPSCs by increasing the expression of TWIST-1/RUNEX-2/ALP signaling axis. PBM therapy notably played a role in the DPSCs genes expression that mediate inflammation process and tissue mineralization. The present data opens a new perspective for PBM therapy in mineralized dental tissue physiology.

Performance of MDP-based system in eroded and carious dentin associated with proteolytic inhibitors: 18-Month exploratory study.

The aim of this study was to explore the impact of the interaction between an MDP-based universal adhesive system in etch-and-rinse mode and two proteolytic inhibitors on the longevity of restorations bonded to artificially-affected-dentin substrates. 90 sound human third molars were randomly distributed into three groups according to the substrate: N-no challenges-control (stored in artificial saliva), ACD-artificial caries dentin (6 h DE + 18 h-RE/5 days + 48 h RE) and ERO-artificial erosion dentin (3 × 5 min/5 days with orange juice). They were further redistributed according to dentin pretreatment: W- water (control), CHX-2% digluconate chlorhexidine and E64- 5 µM E64-Trans-Epoxysuccinyl-L-Leucylamido-(4-guanidino) butane, which resulted in the following 9 groups (n = 10): N-W, N-CHX, N-E64, ACD-W, ACD-CHX, ACD-E64, ERO-W, ERO-CHX and ERO-E64. All specimens were restored with Adper Single Bond Universal (Etch-and-rinse mode)/Filtek Z250. Sticks (0.64 mm2) were obtained and subjected to microtensile test (µTBS) in a universal testing machine at 0.5 mm/min for 7-days, 6 and 18-month analyses. Failure modes were classified using optical microscopy (40X). Data were statistically analyzed by three-way ANOVA and Tukey tests (p < 0.05). All individual factors (p < 0.0001) and interaction between factors were statistically significant (substrate X pretreatment (p = 0.00093); substrate X time (p = 0.01035) and pretreatment X time (p = 0.0035). Caries-affected substrate was the most compromised one, disregarding the pretreatment. CHX was mostly affected compared with E64 up to 18 months, possibly due to its calcium-dependent mechanism.

Profile of a 10-MDP-based universal adhesive system associated with chlorhexidine: Dentin bond strength and in situ zymography performance.

The incorporation of functional monomers and proteolytic inhibitors into adhesive systems have shown to be promising strategies to improve the longevity of adhesive restorations. The aim of this study was to evaluate the long-term bonding performance and anti-gelatinolytic effect of a 10-MDP-based universal adhesive system applied in combination with 2% chlorhexidine digluconate (CHX). For that, this study assessed the resin-dentin bond strength and the in situ gelatinolytic activity profile at the adhesive interface at initial and after 6 month of storage. One hundred and two sound human third molars were prepared and randomly divided into 3 groups according to the adhesive strategy: SB (two-step etch-and-rinse adhesive, Adper Single Bond 2, 10-MDP-free control group); SU-ER (Adper Single Bond Universal, 10-MDP containing universal adhesive applied on etch-and-rinse mode); and SU-SE (SU applied on self-etching mode). The groups were subdivided into two according to the dentin pretreatment: W - water or CHX- 2% chlorhexidine digluconate aqueous solution (SB-W; SB-CHX; SU-ER-W; SU-ER-CHX; SU-SE-W; SU-SE-CHX) and subsequently restored according to the manufacturer's instructions. Bond strength (n = 12) was assessed by a microtensile test (µTBS) (500N/0.5 mm/min) after 24h or after 6 months of storage. In situ zymography was performed to evaluate anti-gelatinolytic activity (n = 5). Resin-dentin samples were incubated with fluorescein-conjugated gelatin for 24 h at 37 °C and analyzed by confocal laser scanning microscopy. Fluorescence indicating gelatinolytic activity at hybrid layer zone and adjacencies was quantified using Image J. Data was analyzed by three-way ANOVA and Tukey post-hoc tests (p < 0.05). Results: SU-SE showed the highest bond strength values, while similar results were observed for SU-ER and SB. No statistical significant differences were observed between pretreatment (CHX vs. W) or storage time (initial vs. 6 months of aging). For in situ zymography, fluorescence was detected in all groups and CHX pre-treatment was able to inhibit the gelatinolytic activity in all conditions. The 10-MDP-based universal adhesive system in self-etching mode was the strategy that showed the best bonding performance irrespective of its combination with chlorhexidine. Pre-treatment with CHX did not impair the bond strength when used in combination with 10-MDP and it may promote collagen stability overtime.

Effect of protease inhibitor specificity on dentin matrix properties.

OBJECTIVE: To evaluate protease activity of dentin matrices subjected to treatment with non-specific (chlorhexidine - CHX), cysteine cathepsin specific (E-64), and cysteine cathepsin-K (CT-K) specific (Odanacatib - ODN) inhibitors. METHODS: Pulverized dentin powder obtained from human dentin disks (0.5 mm thickness) completely demineralized with 10% H3PO4 were challenged in 1 mL lactic acid (LA) (0.1M, pH 5.5) or stored in deionized water for 30 min. Aliquots of dentin powder were then immersed in 1 mL of CHX (2%), E-64 (10 µM and 20 µM) or Odanacatib (0.2 nM and 1 µM) for 30min. Degradation of dentin collagen was determined by telopeptide assays measuring the sub-product release of C-terminal cross-linked telopeptides (ICTP) and C-terminal peptide (CTX) in incubation media, which correlates with matrix metalloproteinases (MMP) and CT-K activities respectively (n = 3). The ICTP and CTX data were normalized to concentration of total protein (ICTPtp and CTXtp) in the media, measured by bicinchoninic acid assay. Dentin matrix properties were also measured by gravimetric change (n = 8) and ultimate tensile strength (UTS) (n = 10). Data were analyzed by one-way ANOVA followed by Tukey's post-hoc test and independent t-test (α = 5%). RESULTS: Telopeptide assays showed significantly lower CTXtp values after treatment with E-64 and Odanacatib. E-64 and Odanacatib at all tested concentrations significantly reduced the release of ICTPtp. Gravimetric analysis showed no significant difference between the tested inhibitors and control except for CHX after lactic acid challenge. UTS results showed significantly higher values for E-64 (20 µM) and Odanacatib (0.2 nM and 1 µM) groups in deionized water. SIGNIFICANCE: Dentin therapies targeting enzymes such as CT-K by specific inhibitors may provide superior pharmacokinetics and optimum efficacy due to precise protein binding, consequently limiting collagen degradation directly or indirectly by enzyme related pathways.

Profiling cytokine levels in chlorhexidine and EGCG-treated odontoblast-like cells.

OBJECTIVE: To screen the effect of two compounds, chlorhexidine diacetate (CHX) and epigallocatechin-gallate (EGCG), on the levels of cytokines produced by odontoblast-like cells (MDPC-23). METHODS: Cells were seeded at 24h and 48h with serial dilution of the compounds to determine cell metabolic activity by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay (n=3). Cells with no compound treatment were used as control (Ctr). For the highest equal non-cytotoxic compound dilution tested at 48h cell treatment, total protein concentration was measured using a Pierce bicinchoninic acid (BCA) assay (n=3), and expression of 23 cytokines was analyzed using the Bio-Plex cytokine assay (n=2). Data were analyzed by one-way ANOVA and Tukey's test (α=5%). RESULTS: The MTT assay revealed that at 24h and 48h, CHX and EGCG did not reduce cell metabolic activity at concentrations of 2.5-20µM (CHX) and 2.5-160µM (EGCG), respectively (p>0.05). At 48h, total protein levels were consistent across all groups for 20µM compound dilution (Ctr: 1.04mg/mL; CHX: 0.98mg/mL; and EGCG: 1.06mg/mL). At 20µM dilution, both CHX and EGCG significantly increased the secretion of IL-1ß, IL-10, IL-12, KC, MIP-1α, IFN-γ and IL-6 (p<0.05). Treatment with CHX significantly increased secretion of IL-4 and RANTES (p<0.05). TREATMENT: with EGCG significantly increased Eotaxin secretion (p<0.05). Both CHX and EGCG significantly decreased secretion of IL-17 (p<0.05). GM-CSF and TNF-α did not present significant change in secretion after treatment with either CHX or EGCG (p>0.05). SIGNIFICANCE: Both CHX and EGCG modulate secretion of various inflammatory and anti-inflammatory mediators in odontoblastic cells.

Co-distribution of cysteine cathepsins and matrix metalloproteases in human dentin.

It has been hypothesized that cysteine cathepsins (CTs) along with matrix metalloproteases (MMPs) may work in conjunction in the proteolysis of mature dentin matrix. The aim of this study was to verify simultaneously the distribution and presence of cathepsins B (CT-B) and K (CT-K) in partially demineralized dentin; and further to evaluate the activity of CTs and MMPs in the same tissue. The distribution of CT-B and CT-K in sound human dentin was assessed by immunohistochemistry. A double-immunolabeling technique was used to identify, at once, the occurrence of those enzymes in dentin. Activities of CTs and MMPs in dentin extracts were evaluated spectrofluorometrically. In addition, in situ gelatinolytic activity of dentin was assayed by zymography. The results revealed the distribution of CT-B and CT-K along the dentin organic matrix and also indicated co-occurrence of MMPs and CTs in that tissue. The enzyme kinetics studies showed proteolytic activity in dentin extracts for both classes of proteases. Furthermore, it was observed that, at least for sound human dentin matrices, the activity of MMPs seems to be predominant over the CTs one.

Effect of sealing infected dentin with glass ionomer cement on the abundance and localization of MMP-2, MMP-8, and MMP-9 in young permanent molars in vivo.

BACKGROUND: The study of MMPs' behavior in carious lesions contributes to the understanding of the mechanisms involved in dentin reorganization after restoration. AIM: To compare the abundance and localization of MMPs 2, 8, and 9 in infected dentin before and after restoration. DESIGN: The sample consisted of 23 young permanent molars with active deep carious lesions. Infected carious dentin samples were collected from the same tooth at baseline and 60 days after cavity lining with GIC and composite resin restoration and processed for immunohistochemistry assays. After digital images were obtained, two calibrated operators analyzed the samples according to the immunostaining intensity and the MMPs' localization. Chi-square test was used for statistical analysis. RESULTS: The intensity of immunostaining for MMP-8 was reduced after 60 days (P = 0.02), and no difference was observed for MMP-2 (P = 0.32) and MMP-9 (P = 0.14). The MMPs' distribution was generalized in the intertubular dentin and absent or located in the intratubular dentin, regardless of the period. CONCLUSION: The sealing of infected carious dentin in young permanent molars reduced the expression of MMP-8, which is consistent with the initial remodeling process of the dentin matrix.

Chlorhexidine does not improve but preserves bond strength to eroded dentin.

PURPOSE: To evaluate the effect of aqueous solutions of chlorhexidine digluconate (CHX) in different concentrations on bond strength to eroded dentin up to 6 months, using normal dentin as a control. METHODS: Exposed flat dentin of extracted third molars was only ground with 600-grit SiC paper/1 minute (normal dentin - N), or subsequently eroded by a regular-cola soft-drink (eroded dentin - E). N and E were acid-etched, washed, dried and rehydrated with 1.5 µL, respectively, of distillated water (control - NC / EC); of 0.004% CHX (N0.004% / E0.004%); or of 2% CHX (N2% / E2%). Adper Single Bond 2 was applied in all specimens and resin composite buildups were constructed with Filtek Z350. Specimens were sectioned in beams, which were tested (µTBS) immediately or after 6 months of aging. RESULTS: Microtensile bond strength to eroded dentin was always significantly lower than that to normal dentin. Application of tested CHX solutions did not exert a significant effect immediately; however, after aging, the 2% CHX prevented abrupt bond strength loss both to eroded and normal dentin.

Transdentinal cytotoxicity of experimental adhesive systems of different hydrophilicity applied to ethanol-saturated dentin.

The aim of this study was to evaluate the transdentinal cytotoxicity of experimental adhesive systems (EASs) with different hydrophilicity and dentin saturation solutions on odontoblast-like cells. One hundred 0.4-mm-thick dentin discs were mounted in in vitro pulp chambers and assigned to 10 groups. MDPC-23 cells were seeded onto the pulpal side of the discs, incubated for 48h. The EASs with increasing hydrophilicity (R1, R2, R3 and R4) were applied to the occlusal side after etching and saturation of etched dentin with water or ethanol. R0 (no adhesive) served as controls. R1 is a non-solvated hydrophobic blend, R2 is similar to a simplified etch-and-rinse adhesive system and R3 and R4 are similar to self-etching adhesives. After 24h, cell metabolism was evaluated by MTT assay (n=8 discs) and cell morphology was examined by SEM (n=2 discs). Type of cell death was identified by flow cytometry and the degree of monomer conversion (%DC) was determined by infrared spectroscopy (FTIR) after 10s or 20s of photoactivation. Data were analyzed by the Kruskal-Wallis and Mann-Whitney tests (α=0.05). Dentin saturation with ethanol resulted in higher necrotic cell death ratios for R2, R3 and R4 compared with water saturation, although R2 and R3 induced higher SDH production. Photoactivation for 20s significantly improved the %DC of all EASs compared with 10s. A significant positive correlation was observed between the degree of hydrophilicity and %DC. In conclusion, except for R1, dentin saturation with ethanol increased the cytotoxicity of EASs, as expressed by the induction of necrotic cell death.

Cytotoxicity of adhesive systems of different hydrophilicities on cultured odontoblast-like cells.

This study evaluated the cytotoxicity of experimental adhesive systems (EASs) on odontoblast-like cells. Paper discs (n = 132) were impregnated with 10 µL of each EAS-R1, R2, R3, R4, and R5 (in an ascending order of hydrophilicity), followed by photoactivation. R1 and R2 are nonsolvated hydrophobic blends, R3 represents a simplified etch-and-rinse adhesive system, and R4 and R5 represent simplified self-etch adhesive systems. Discs were immersed in Dulbecco's modified Eagle's medium for 24 h to obtain eluates applied on MDPC-23 cell cultures. No material was applied on discs used as control (R0). Cell viability [3-(4,5-dimethythiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay], total protein (TP) production, alkaline phosphatase (ALP) activity, type of cell death, and degree of monomer conversion Fourier transform infrared (%DC-FTIR) were evaluated. Data were analyzed by Kruskal-Wallis and Mann-Whitney tests (α = 0.05). Considering R0 (control) as having 100% of cell viability, R1, R2, R3, R4, and R5 reduced the metabolic activity of cells by 36.4, 3.1, 0.2, 21.5, and 65.7%, respectively, but only R1 and R5 differed from R0. Comparing with R0, lower TP production was observed for R1, R4, and R5, while ALP activity decreased for R1 and R5. Necrotic cell death was predominant for all EASs, but only R1, R4, and R5 differed from R0. Only R5 presented a different apoptotic cell death ratio from R0. R1 presented the lowest %DC (ca. 37%), whereas R4 and R5 presented the highest (ca. 56%). In conclusion, R2 and R3 were not toxic to the MDPC-23 cells, suggesting that the degree of hydrophilicity or %DC of the EASs alone were not responsible for their cytopathic effects.

Microtensile bond strength test and failure analysis to assess bonding characteristics of different adhesion approaches to ground versus unground enamel.

This study evaluated the bonding characteristics to ground and unground enamel obtained with different strategies. For this purpose, 24 sound third-molars were bisected mesiodistally to obtain tooth halves. A flat enamel area was delimited in the tooth sections, which were randomly distributed into 8 groups (n=6), according to the enamel condition (ground and unground) and adhesive system (Adper Single Bond 2 - SB2; Adper Prompt L-Pop - PLP; Adper Prompt - AD; Clearfil SE Bond - SE). Each system was applied according manufacturers' instructions and a 6-mm-high resin composite "crown" was incrementally built up on bonded surfaces. Hourglass-shaped specimens with 0.8 mm(2) cross-section were produced. Microtensile bond strength (µTBS) was recorded and the failure patterns were classified. Results were analyzed by two-way ANOVA and Tukey's test (α=0.05). There were no statistically significant differences among the µTBS values of SB2, PLP and AD (p>0.05). SE values were significantly lower (p0.05). There was prevalence of cohesive failure within enamel, adhesive system and resin composite for SB2. The self-etch systems produced higher incidence of cohesive failures in the adhesive system. Enamel condition did not determine significant differences on bonding characteristics for the same bonding system. In conclusion, the bonding systems evaluated in this study resulted in specific µTBS and failure patterns due to the particular interaction with enamel.

Microtensile bond strength test and failure analysis to assess bonding characteristics of different adhesion approaches to ground versus unground enamel

This study evaluated the bonding characteristics to ground and unground enamel obtained with different strategies. For this purpose, 24 sound third-molars were bisected mesiodistally to obtain tooth halves. A flat enamel area was delimited in the tooth sections, which were randomly distributed into 8 groups (n=6), according to the enamel condition (ground and unground) and adhesive system (Adper Single Bond 2 - SB2; Adper Prompt L-Pop - PLP; Adper Prompt - AD; Clearfil SE Bond - SE). Each system was applied according manufacturers' instructions and a 6-mm-high resin composite "crown" was incrementally built up on bonded surfaces. Hourglass-shaped specimens with 0.8 mm² cross-section were produced. Microtensile bond strength (μTBS) was recorded and the failure patterns were classified. Results were analyzed by two-way ANOVA and Tukey's test (α=0.05). There were no statistically significant differences among the μTBS values of SB2, PLP and AD (p>0.05). SE values were significantly lower (p0.05). There was prevalence of cohesive failure within enamel, adhesive system and resin composite for SB2. The self-etch systems produced higher incidence of cohesive failures in the adhesive system. Enamel condition did not determine significant differences on bonding characteristics for the same bonding system. In conclusion, the bonding systems evaluated in this study resulted in specific μTBS and failure patterns due to the particular interaction with enamel.

Este estudo avaliou a união ao esmalte íntegro e desgastado obtida com diferentes estratégias. Para tanto, 24 terceiros molares hígidos foram seccionados ao meio. Um plano de esmalte foi delimitado nos fragmentos de dente, aleatoriamente distribuídos em 8 grupos (n=6) conforme a condição do esmalte (íntegro ou desgastado) e o adesivo (Adper Single Bond 2: SB2; Adper Prompt L-Pop: PLP; Adper Prompt: AD; Clearfil SE Bond: SE). Estes foram aplicados seguindo recomendações dos fabricantes e uma "coroa" de compósito (altura - 6 mm) incrementalmente construída. Espécimes (ampulheta/secção transversal - 0,8 mm²) foram confeccionados. O ensaio de resistência da união à micro-tração (RUµT) foi realizado e os padrões de fratura classificadas. Os resultados analisados pela ANOVA (dois fatores) e teste de Tukey (α=0,05). Os valores de RUµT do SB2, PLP e AD não foram significativamente diferentes entre si (p>0,05); SE foi inferior (p0,05). Houve prevalência de fratura no esmalte, adesivo e compósito no SB2. Nos adesivos autocondicionantes, predominou a fratura no adesivo. A condição do esmalte não influenciou significativamente as características da união, considerando o mesmo adesivo. Em conclusão, os adesivos apresentaram RUµT e padrões de fratura específicos, devido a forma de interação com o esmalte.

Effect of oxalate desensitizer on the durability of resin-bonded interfaces.

Potassium oxalate desensitizers were previously shown to effectively reduce the immediate permeability of resin-bonded dentin. The current study evaluated whether the effect of the combined application of oxalate with etch-and-rinse adhesives interferes with the durability of resin-dentin bonds when using etch-and-rinse adhesives. The bond strength of resin-bonded dentin specimens composed of two-step or three-step etch-and-rinse adhesives (Single Bond, One-Step and Scotchbond Multi-Purpose, respectively) was tested immediately (24 hours) and after 12 months of water storage. The adhesives were used either according to the manufacturers' instructions (control groups) or after treating acid-etched dentin with a potassium oxalate gel (BisBlock, BISCO, Inc). The treatment of dentin with potassium oxalate was shown to negatively affect the baseline bond strength of resin-bonded dentin specimens, regardless of the adhesive used (p < 0.05). After storage, the bond strength of the resin-bonded interfaces was significantly reduced for all the tested groups (p < 0.001). Nevertheless, the rate of decreasing bond strength was significantly lower for oxalate-treated specimens than for the controls (p < 0.05).

Influences of surface and solvent on retention of HEMA/mixture components after evaporation.

OBJECTIVES: This study examined the retention of solvents within experimental HEMA/solvent primers after two conditions for solvent evaporation: from a free surface or from dentine surface. METHODS: Experimental primers were prepared by mixing 35% HEMA with 65% water, methanol, ethanol or acetone (v/v). Aliquots of each primer (50 microl) were placed on glass wells or they were applied to the surface of acid-etched dentine cubes (2mm x 2mm x 2mm) (n=5). For both conditions (i.e. from free surface or dentine cubes), change in primers mass due to solvent evaporation was gravimetrically measured for 10min at 51% RH and 21 degrees C. The rate of solvent evaporation was calculated as a function of loss of primers mass (%) over time. Data were analysed by two-way ANOVA and Student-Newman-Keuls (p<0.05). RESULTS: There were significant differences between solvent retention (%) and evaporation rate (%/min) depending on the solvent present in the primer and the condition for evaporation (from free surface or dentine cubes) (p<0.05). For both conditions, the greatest amount of retained solvent was observed for HEMA/water primer. The rate of solvent evaporation for HEMA/acetone primer was almost 2- to 10-times higher than for HEMA/water primer depending whether evaporation occurred, respectively, from a free surface or dentine cubes. The rate of solvent evaporation varied with time, being in general highest at the earliest periods. CONCLUSIONS: The rate of solvent evaporation and its retention into HEMA/solvent primers was influenced by the type of the solvent and condition allowed for their evaporation.

Effect of an additional hydrophilic versus hydrophobic coat on the quality of dentinal sealing provided by two-step etchand-rinse adhesives

OBJECTIVE: To test the hypothesis that the quality of the dentinal sealing provided by two-step etch-and-rinse adhesives cannot be altered by the addition of an extra layer of the respective adhesive or the application of a more hydrophobic, non-solvated resin. MATERIAL AND METHODS: full-crown preparations were acid-etched with phosphoric acid for 15 s and bonded with Adper Single Bond (3M ESPE), Excite DSC (Ivoclar/Vivadent) or Prime & Bond NT (Dentsply). The adhesives were used according to the manufacturers' instructions (control groups) or after application to dentin they were a) covered with an extra coat of each respective system or b) coated with a non-solvated bonding agent (Adper Scotchbond Multi-Purpose Adhesive, 3M ESPE). Fluid flow rate was measured before and after dentin surfaces were acid-etched and bonded with adhesives. RESULTS: None of the adhesives or experimental treatments was capable to block completely the fluid transudation across the treated dentin. Application of an extra coat of the adhesive did not reduce the fluid flow rate of adhesive-bonded dentin (p>0.05). Conversely, the application of a more hydrophobic non-solvated resin resulted in significant reductions in the fluid flow rate (p<0.05) for all tested adhesives. CONCLUSIONS: The quality of the dentinal sealing provided by etch-and-rinse adhesives can be significantly improved by the application of a more hydrophobic, non-solvated bonding agent.

Effect of an additional hydrophilic versus hydrophobic coat on the quality of dentinal sealing provided by two-step etch-and-rinse adhesives.

OBJECTIVE: To test the hypothesis that the quality of the dentinal sealing provided by two-step etch-and-rinse adhesives cannot be altered by the addition of an extra layer of the respective adhesive or the application of a more hydrophobic, non-solvated resin. MATERIAL AND METHODS: full-crown preparations were acid-etched with phosphoric acid for 15 s and bonded with Adper Single Bond (3M ESPE), Excite DSC (Ivoclar/Vivadent) or Prime & Bond NT (Dentsply). The adhesives were used according to the manufacturers' instructions (control groups) or after application to dentin they were a) covered with an extra coat of each respective system or b) coated with a non-solvated bonding agent (Adper Scotchbond Multi-Purpose Adhesive, 3M ESPE). Fluid flow rate was measured before and after dentin surfaces were acid-etched and bonded with adhesives. RESULTS: None of the adhesives or experimental treatments was capable to block completely the fluid transudation across the treated dentin. Application of an extra coat of the adhesive did not reduce the fluid flow rate of adhesive-bonded dentin (p>0.05). Conversely, the application of a more hydrophobic non-solvated resin resulted in significant reductions in the fluid flow rate (p<0.05) for all tested adhesives. CONCLUSIONS: The quality of the dentinal sealing provided by etch-and-rinse adhesives can be significantly improved by the application of a more hydrophobic, non-solvated bonding agent.

Microtensile bond strength of adhesive systems to dentin with or without application of an intermediate flowable resin layer.

This study evaluated the effect of flowable composite resin application on the microtensile bond strength (microTBS) of adhesive systems to dentin. Occlusal surfaces of human third molars were ground to obtain flat dentin surfaces. The crown of each tooth was sectioned occluso-gingivally into four quarters with a water-cooled diamond saw. One of the following adhesive systems was applied to dentin surface in each quarter of the same tooth, following manufacturers' instructions: Scotchbond Multipurpose, Single Bond Adper Prompt and Clearfil SE Bond. Experimental and control groups received the same treatment, except for the fact that a layer of flowable composite (Filtek Flow) was placed and light-cured on top of the adhesive layer in the specimens of the experimental groups. Resin composite (Filtek Z250) crown buildups were then made on the bonded surfaces and incrementally light-cured for 20 s. The restored teeth were stored in water at 37 degrees C for 24 h. Each tooth quarter was serially cut in a longitudinal direction in order to obtain several bonded sticks (0.9-mm(2) in cross-section). Maximal microtensile stress (in MPa) at failure was recorded using a universal testing machine. Data were analyzed by two-way ANOVA and Tukey's test at 5% significance level. Placement of a low-viscosity resin to the hybridized dentin increased the microTBS for all tested adhesive systems. However, such increase was significant only for Clearfil SE Bond (p<0.05). When the restorations were placed without low-viscosity resin, Clearfil SE Bond presented higher microTBS than Adper Prompt (p<0.05). For the groups treated with flowable composite, Clearfil SE Bond produced the highest microTBS means to dentin. In conclusion, the effect of the addition of an intermediate flowable composite layer on microTBS to dentin was material-dependent and resulted in an increased adhesion for all tested materials, though with significance only for Clearfil SE Bond.

Microtensile bond strength of adhesive systems to dentin with or without application of an intermediate flowable resin layer

This study evaluated the effect of flowable composite resin application on the microtensile bond strength (muTBS) of adhesive systems to dentin. Occlusal surfaces of human third molars were ground to obtain flat dentin surfaces. The crown of each tooth was sectioned occluso-gingivally into four quarters with a water-cooled diamond saw. One of the following adhesive systems was applied to dentin surface in each quarter of the same tooth, following manufacturers' instructions: Scotchbond Multipurpose, Single Bond Adper Prompt and Clearfil SE Bond. Experimental and control groups received the same treatment, except for the fact that a layer of flowable composite (Filtek Flow) was placed and light-cured on top of the adhesive layer in the specimens of the experimental groups. Resin composite (Filtek Z250) crown buildups were then made on the bonded surfaces and incrementally light-cured for 20 s. The restored teeth were stored in water at 37ºC for 24 h. Each tooth quarter was serially cut in a longitudinal direction in order to obtain several bonded sticks (0.9-mm² in cross-section). Maximal microtensile stress (in MPa) at failure was recorded using a universal testing machine. Data were analyzed by two-way ANOVA and Tukey's test at 5 percent significance level. Placement of a low-viscosity resin to the hybridized dentin increased the muTBS for all tested adhesive systems. However, such increase was significant only for Clearfil SE Bond (p<0.05). When the restorations were placed without low-viscosity resin, Clearfil SE Bond presented higher muTBS than Adper Prompt (p<0.05). For the groups treated with flowable composite, Clearfil SE Bond produced the highest muTBS means to dentin. In conclusion, the effect of the addition of an intermediate flowable composite layer on muTBS to dentin was material-dependent and resulted in an increased adhesion for all tested materials, though with significance only for Clearfil SE Bond.

Este estudo avaliou o efeito da aplicação de uma resina flow na resistência de união (RU) (microtração) de sistemas adesivos à dentina. Superfícies oclusais de terceiros molares humanos foram abrasionadas para formar superfícies dentinárias planificadas. As coroas foram seccionadas no sentido ocluso-gengival em quatro partes de tamanhos similares com disco diamantado sob refrigeração. Os sistemas adesivos (Scotchbond Multipurpose; Single Bond; Adper Prompt e Clearfil SE Bond) foram aplicados nas superfícies dentinárias, de acordo com as instruções dos fabricantes, sendo cada um deles em uma das quatro partes de um mesmo dente. Os grupos experimentais e controle receberam o mesmo tratamento, porém a resina flow (Filtek Flow) foi aplicada e fotoativada sobre os sistemas adesivos nos espécimes dos grupos experimentais. Um bloco de compósito (Filtek Z250) foi confeccionado em incrementos sobre as superfícies tratadas com o sistema adesivo e fotoativado por 20 s. Os dentes restaurados foram armazenados em água a 37ºC por 24 h e seccionados longitudinalmente para obtenção de espécimes com formato de paralelepípedo, com 0,9 mm² na área de união. O ensaio de tração foi realizado em máquina universal de ensaios e os valores de RU foram expressos em MPa. Os dados foram analisados pela ANOVA (dois fatores) e teste de Tukey (5 por cento). A aplicação da resina flow na dentina previamente hibridizada aumentou a RU para todos os sistemas adesivos, entretanto, foi estatisticamente significativa somente para o adesivo Clearfil SE Bond (p<0,05). Quando as restaurações foram confeccionadas sem a resina de baixa viscosidade, o Clearfil SE Bond obteve maior valor de RU que o adesivo Adper Prompt (p<0,05). Para os grupos com a resina flow, o Clearfil SE Bond resultou no maior valor médio de RU à dentina. Conclui-se que o efeito da aplicação da resina flow foi depende do sistema adesivo utilizado e indicou um aumento na RU para todos os sistemas adesivos...

Water sorption and solubility of dentin bonding agents light-cured with different light sources.

OBJECTIVE: The aim of this study was to compare water sorption (WS) and solubility (WSB) of different dentin bonding agents (DBA) as regards classification and light-activation system. The null hypotheses were: (1) there is no difference among DBA with respect to water sorption and solubility; (2) there is no effect of light-curing source on water sorption and solubility of DBA. METHODS: The tested materials were: three-step etch-and-rinse (ScotchBond multi-purpose and Heliobond-control groups), two-step etch-and-rinse (Excite, Adper Single Bond, Adper Single Bond 2), self-etching (Adhse) and all-in-one (Xeno III) systems. For each material, six specimens were prepared (0.8mmx5.5mm) for each light-activation device: halogen lamp (control) or light-emitting diodes (LED). They were transferred to desiccators until a constant mass was obtained (m(1)), subsequently immersed in deionized water until no alteration was obtained (m(2)). Following, the specimens were reconditioned to constant mass in desiccators (m(3)). The volume (V) of each specimen was calculated (mm(3)). WS and WSB were determined using the equations (m(2)-m(3)/V) and (m(3)-m(1)/V), respectively. Data were subjected to one-way analysis of variance and Tukey test (p<0.05). The influence of photo-activation units was analyzed by t-test. RESULTS: Higher values of WS and WSB were observed for the two-step etch-and-rinse and all-in-one systems. The employed light-activation unit also affected the tested characteristics. Thus, the anticipated null hypotheses were rejected. CONCLUSIONS: Simplified systems were more susceptible to water adverse and they may become more prone to degradation over time regardless of the type of light-activation source.

Effects of a combined application of potassium oxalate gel/adhesive agent on dentin permeability in vitro.

PURPOSE: To test the effects of sequential application of potassium oxalate gel/adhesive agent on in vitro dentin permeability. MATERIALS AND METHODS: Full crown preparations were made in extracted human molars to expose deep coronal dentin. The roots and pulp were removed and the resulting crown segments were connected to a special device (Flodec) to permit the measurement of the permeability of the specimens before and after treatments. Minimum and maximum permeability were recorded after smear layer and phosphoric acid treatment. A new smear layer was created and the permeability measured after the crowns were bonded with Single Bond (3M ESPE), One-Up Bond F (Tokuyama), and AdheSE (Ivoclar Vivadent), either according to manufacturer's instructions or after treating the acid-etched dentin with a 3 wt% potassium oxalate gel. The results were expressed as a percentage of maximum permeability values. Impressions and epoxy resin replicas from the crown segments were produced for SEM examination. RESULTS: None of the adhesives were able to eliminate the fluid flow through dentin. Two-way ANOVA revealed that the application of potassium oxalate prior to the bonding procedures was the most effective technique in reducingthe dentin permeability (p < 0.05), regardless of the adhesive used. SEM micrographs showed that transudation of dentinal fluid could be identified on the surfaces of all replicas. CONCLUSION: The use of potassium oxalate gel was effective in reducing the permeability of bonded dentin.