Universal adhesive: the effect of different simulated pulpal pressure fluids and bonding modes to dentin.

The aim of this study was to evaluate the effect of SPP with either fetal bovine serum (FBS) or deionized water (DW) on the bond strength (µTBS) of a Universal adhesive to dentin, in both etch-and-rinse (ER) and self-etch (SE) modes. The kinematic viscosity (cSt) of FBS and DW was measured at 25 °C ± 0.1 ºC. Seventy-two sound human molars were sectioned and randomly divided into three groups according to the SPP conditions: (1) Control (0 cm H2O), (2) SPP (15 cm H2O) with FBS, (3) SPP (15 cm H2O) with DW. Each group was subdivided (n = 10) based on the bonding modes: ER (37% phosphoric acid + ScothBond Universal Adhesive) or SE (ScothBond Universal Adhesive). Samples were then submitted to µTBS. Data were analyzed by Student's t test, two-way ANOVA and Tukey tests (p < 0.05). The cSt results showed that DW (23.59 ± 0.39) had significantly higher values than FBS (22.33 ± 0.06). With regard to SPP, the control group (36.1 MPa) had significantly higher values of µTBS when compared to the SPP using FBS (31.06 MPa) and SPP with DW (26.55 MPa). According to ANOVA, the bonding modes and the interaction of simulated pulpal pressure (SPP) did not statistically influence the results (p < 0.05). The presence of SPP reduced the bond strength of Universal adhesive to dentin. DW during SPP had significantly reduced bonding values when compared to FBS. Bonding strategies were not affected by SPP when evaluated in a short period of time (24 h).

Bond Strength of Universal Self-Etch 1-Step Adhesive Systems for Orthodontic Brackets.

OBJECTIVE: The objective of this study was to assess the shear bond strength (SBS) of orthodontic brackets bonded to uncut enamel with universal self-etch 1-step adhesive systems. METHODS: Extracted uncut premolars (n = 160) were randomly divided into 4 groups for treatment with Scotchbond Universal Adhesive (SU), All-Bond Universal (BU), Clearfil Universal Bond (CU) or the control, Adper Scotchbond Multi-Purpose Adhesive. Following bonding of brackets on tooth surfaces, teeth were stored in distilled water for 24 h and 6 months, and brackets were tested for SBS. The adhesive remnant index (ARI) and quantitative percentage of remaining resin (%RR) were recorded. Scanning electron microscopy was used to analyze debonded surfaces qualitatively. SBS and %RR data were analyzed by 2-way ANOVA followed by the Tukey test (α = 0.05). RESULTS: At neither time did these universal adhesives achieve satisfactory SBS for orthodontic treatment. The control group had the highest SBS, ARI score and mean %RR (and these differences were significant), while the BU group had the lowest SBS. SBS mean values and ARI scores decreased over time for SU and BU, but remained stable for CU. There was no difference in %RR among the universal adhesives tested. CONCLUSION: None of the universal adhesives used in self-etch mode achieved SBS values (at 24 h and 6 months) that were satisfactory for orthodontic treatment.

Effect of different concentrations and application times of proanthocyanidin gels on dentin erosion.

PURPOSE: To analyze the effect of different concentrations and application times of proanthocyanidin gels on dentin before an erosive challenge in order to evaluate if there is a dose-response or application time-response relationship in the use of these gels for erosion prevention. METHODS: 80 bovine root dentin blocks were randomly and equally divided into 10 groups and treated according to the two factors under study (purified grape seed proanthocyanidin gel concentration and time of application): 0.05P1: 0.05% proanthocyanidin gel during 1 minute; 0.05P5: 0.05% proanthocyanidin gel during 5 minutes; 1P1: 1% proanthocyanidin gel during 1 minute; 1P5: 1% proanthocyanidin gel during 5 minutes; 5P1: 5% proanthocyanidin gel during 1 minute; 5P5: 5% proantho-cyanidin gel during 5 minutes; 10P1: 10% proanthocyanidin gel during 1 minute; 10P5: 10% proanthocyanidin gel during 5 minutes; Control 1: placebo gel during 1 minute; and Control 5: placebo gel during 5 minutes. The gels were applied over dentin blocks once before the first erosive challenge. After that, the blocks were subjected to three erosive cycles per day, during 5 days. Profilometry was used to quantify the dentin loss (µm). Data were analyzed by two-way ANOVA and Fisher's test (P< 0.05). RESULTS: Statistical analysis showed that there was no significant difference between the application times. The different concentrations of proanthocianidin gels presented similar results (P> 0.05). All tested gels resulted in significantly less wear when compared to the placebo gel. CLINICAL SIGNIFICANCE: Grape seed proanthocyanidin gels could be considered as a promising therapy to diminish erosive dentin wear because it may interact with the exposed collagen, enhancing the demineralized organic matrix stabilization, which acts as a barrier against the diffusion of the acids from erosion.

Chemical and Ultrastructural Characterization of Dentin Treated with Remineralizing Dentifrices.

The aim of this study is to investigate dentin chemical and ultrastructural changes upon exposure to remineralizing dentifrices. Dentin disks were obtained from permanent human molars and treated for 7 days with the dentifrices: (1) C group-control (no dentifrice); (2) S group-Sensodyne Repair & Protect; (3) D group-Dentalclean Daily Regenerating Gel; and (4) DB group-D group + Dentalclean regenerating booster. Afterwards, samples were submitted to an additional 7 days of toothbrushing associated with daily acidic challenge. Samples were imaged and analyzed (days 1, 7, and 14) for Young's modulus by atomic force microscopy (AFM), scanning electron microscopy (SEM) and energy-dispersive X-ray spectroscopy (EDX), transmission electron microscopy (TEM) and selected area electron diffraction (SAED). SEM and AFM revealed precipitate deposition on dentin surfaces in groups S, D, and DB, formed as early as day 1. Surface elemental analysis showed a Si increase on all brushed surfaces. Similar surface morphology was maintained after the acidic challenge period. Bright-field TEM/SAED revealed the formation of nanocrystalline hydroxyapatite inside the dentin tubules of groups S, D, and DB after day 7. Group C presented a gradual reduction of Young's modulus from days-1-14, whereas all remaining groups had increased values. All evaluated dentifrices led to successful formation of hydroxyapatite and increased dentin stiffness.

Longitudinal bond strength of a universal adhesive and chemical dentin characterization under different acid etching protocols.

OBJECTIVE: This study aimed to analyze the longitudinal bond strength of a universal adhesive and chemically characterize the dentin substrate under different acid etching protocols. METHODOLOGY: Dentin samples were etched with polyacrylic acid 25% (PAA) for 10 seconds (n=3) and phosphoric acid 32% (PA) for 15 seconds (n=3) and analyzed by Fourier transform infrared spectroscopy - attenuated total reflectance (FTIR-ATR) before and after treatment. For collagen degradation, samples (n=12) were divided into 3 groups: PAA, PA, and Deionized water (control), and analyzed by the quantity of solubilized type I collagen C-terminal cross-linked telopeptides and solubilized C-terminal peptide in relation to total protein concentration (ICTPtp and CTXtp) and by their ultimate tensile strength (UTS). For the adhesive interface analysis, dentin samples (n=72) were divided into 3 groups: PAA, PA, and Self-etch (SE), and subdivided into 2 groups: 24 h (baseline) and 1 year. The following tests were performed: microtensile bond strength (µTBS) (n=48), scanning electron microscopy (SEM) (n=12), and nanoleakage (n=12). RESULTS: The FTIR of PAA showed lower reduction of the peaks in the phosphate group when compared to PA. For ICTPtp, PA showed a significantly higher value. For CTXtp, PA and PAA groups failed to statically differ from each other. UTS was significantly lower for PA. For µTBS, storage time significantly affected bond strength. The results were unaffected by the etching protocol. For SEM, after 1 year, PA had little evidence of degradation in the upper third of the adhesive interface in comparison to the other groups. Nanoleakage showed no considerable silver impregnation after 1 year in the SE group. CONCLUSION: The use of PAA prior to a universal adhesive (when compared to PA) represents a less aggressive type of etching to dentin. However, self-etching still seems to be the best option for universal adhesive systems that have functional monomers in their composition.

Gingival microleakage of class V composite restorations with fiber inserts.

AIM: This study investigated the effect of different fiber inserts (glass and polyethylene), bonding agents, and resin composites on the gingival margin microleakage of class V composite restorations. MATERIALS AND METHODS: Sixty premolars were sterilized and mounted in acrylic resin bases. Class V cavities were prepared buccally and lingually, 1 mm below the cementoenamel junction, comprising 12 groups (n = 10). In the experimental groups fiber inserts were cut and placed at the gingival seat, while the control groups had no inserts. Combinations of two composite materials, Filtek-Z250 and Filtek-LS (3M-ESPE), and four bonding agents, Clearfil SE bond (Kuraray) (C), Scotch Bond Multipurpose (3M-ESPE) (SB), Prime and Bond NT (Dentsply) (PB), and Filtek-LS (3M-ESPE) (LS) were used. Restorations were incrementally inserted and polymerized for 40s. Specimens were then stored in distilled water for 7 days and thermocycled for 500 cycles. Teeth surfaces were sealed with nail polish except for 1 mm around restoration margins and immersed in 2% red procion dye. Teeth were then sectioned buccolingually and dye penetration was assessed with five-point scale. Data were statistically-analyzed by Kruskal-Wallis, ANOVA and Tukey's tests (α = 5%). RESULTS: Mean microleakage scores varied from 0.40 (Groups C, C with polyethylene, LS, LS with polyethylene) to 1.50 (SB). CONCLUSION: Different bonding agents led to differences in microleakage scores where C and LS showed significantly lower microleakage than PB. SB had highest mean microleakage score, however, incorporation of fibers resulted in significant reduction in microleakage. CLINICAL SIGNIFICANCE: Class V resin composite restorations bonded with a total-etch adhesive had a significant reduction in mean microleakage scores when glass or polyethylene fibers were placed at the gingival cavo-surface margin. In contrast, for two self-etch adhesive systems, the incorporation of fibers had no significant effect on mean microleakage scores.

Effect of Odanacatib on the release of NTX (Amino Terminal Telopeptide) from LPS contaminated type I dentin collagen.

OBJECTIVE: To evaluated the Odanacatib inhibitor treatment on lipopolysaccharide (LPS) contamination effect on cathepsin-K mediated dentin degradation by analysis of type I collagen C- and N-termini telopeptides. METHODS: Pulverized and disks of human dentin were demineralized and LPS contaminated, or stored in deionized water (DW) for 12 h. Samples were challenged with lactic acid (LA). Aliquots of dentin powder were treated with 1 mL Odanacatib or stored in DW for 30 min. Dentin collagen degradation was determined by sub-product release of C-terminal (ICTP and CTX) and N-terminal (NTX) telopeptides, normalized to total protein (tp) concentration (n = 3). Dentin matrix was evaluated for gravimetric (n = 8) and ultrastructural changes. Data were analyzed by Student t-test, one-way ANOVA and Tukey's test (α = 5 %). RESULTS: LA incubation significantly increased telopeptide release compared with DW (p < 0.05). In untreated groups, significantly higher CTXtp, NTXtp telopeptide rates were observed for LA+LPS samples compared with DW (p < 0.01). Odanacatib significantly reduced ICTPtp, CTXtp, and NTXtp telopeptide release for LPS, LA, and LA+LPS conditions. In untreated groups, LPS and LA+LPS challenge significantly increased dentin weight loss (p = 0.02). Within each storage condition, Odanacatib treatment did not affect weight change (p > 0.05) of dentin disks. SIGNIFICANCE: This study showed that LPS contamination resulted in significantly higher rates of NTX than CTX from dentin matrix. Odanacatib significantly reduced telopeptide release rates of LPS contaminated dentin matrix.

An 8-year follow-up of resin infiltration on anterior white spot lesions.

White spot lesions (WSLs) are sites of enamel surface and subsurface demineralization that increases tissue porosity and affects the teeth appearance. The resin infiltration technique proved to be a valid alternative to arrest caries lesion progression and to masking a color change in noncavitated WSLs. Thus, this study aims to report a clinical case of anterior WSLs treated with resin infiltration technique with an 8-year follow-up. The resin infiltration protocol was performed in an 18-year-old female patient presenting WSLs on the maxillary right lateral incisor, left central incisor, and left canine. The protocol followed the manufacturer's recommendations. The patient reported satisfaction with the smile appearance, at the end of the appointment. Infiltrated areas remained unchanged after an 8-year follow-up, showing an acceptable result for the patient's esthetic desires. After 8 years of evaluation, the resin infiltration technique proved to be a resistant and reliable alternative in preventing caries progression and in color masking WSLs.

Synergistic antimicrobial potential of EGCG and fosfomycin against biofilms associated with endodontic infections.

OBJECTIVE: This study aimed to evaluate the cytotoxicity and synergistic effect of epigallocatechin gallate (EGCG) and fosfomycin (FOSFO) on biofilms of oral bacteria associated with endodontic infections. METHODOLOGY: This study determined minimum inhibitory and bactericidal concentration (MIC/MBC) and fractionated inhibitory concentration (FIC) of EGCG and FOSFO against Enterococcus faecalis, Actinomyces israelii, Streptococcus mutans, and Fusobacterium nucleatum. Monospecies and multispecies biofilms with those bacteria formed in polystyrene microplates and in radicular dentin blocks of bovine teeth were treated with the compounds and control chlorhexidine (CHX) and evaluated by bacterial counts and microscopy analysis. Toxicity effect of the compounds was determined on fibroblasts culture by methyl tetrazolium assays. RESULTS: The combination of EGCG + FOSFO demonstrated synergism against all bacterial species, with an FIC index ranging from 0.35 to 0.5. At the MIC/FIC concentrations, EGCG, FOSFO, and EGCG+FOSFO were not toxic to fibroblasts. EGCG+FOSFO significantly reduced monospecies biofilms of E. faecalis and A. israelli, whereas S. mutans and F. nucleatum biofilms were eliminated by all compounds. Scanning electron microscopy of multispecies biofilms treated with EGCG, EGCG+FOSFO, and CHX at 100x MIC showed evident biofilm disorganization and substantial reduction of extracellular matrix. Confocal microscopy observed a significant reduction of multispecies biofilms formed in dentin tubules with 84.85%, 78.49%, and 50.6% of dead cells for EGCG+FOSFO, EGCG, and CHX at 100x MIC, respectively. CONCLUSION: EGCG and fosfomycin showed a synergistic effect against biofilms of oral pathogens related to root canal infections without causing cytotoxicity.

Cytotoxicity and Biomineralization Potential of Flavonoids Incorporated into PNVCL Hydrogels.

This study aimed to evaluate the effects of flavonoids incorporated into poly(N-vinylcaprolactam) (PNVCL) hydrogel on cell viability and mineralization markers of odontoblast-like cells. MDPC-23 cells were exposed to ampelopsin (AMP), isoquercitrin (ISO), rutin (RUT) and control calcium hydroxide (CH) for evaluation of cell viability, total protein (TP) production, alkaline phosphatase (ALP) activity and mineralized nodule deposition by colorimetric assays. Based on an initial screening, AMP and CH were loaded into PNVCL hydrogels and had their cytotoxicity and effect on mineralization markers determined. Cell viability was above 70% when MDPC-23 cells were treated with AMP, ISO and RUT. AMP showed the highest ALP activity and mineralized nodule deposition. Extracts of PNVCL+AMP and PNVCL+CH in culture medium (at the dilutions of 1/16 and 1/32) did not affect cell viability and stimulated ALP activity and mineralized nodules' deposition, which were statistically higher than the control in osteogenic medium. In conclusion, AMP and AMP-loaded PNVCL hydrogels were cytocompatible and able to induce bio-mineralization markers in odontoblast-cells.

Effect of taxifolin and epigallocatechin-3-gallate on biomineralization potential of stem cells from dental apical papilla.

OBJECTIVE: Considering the variety of pharmacological activities and the potential to mediate biomineralization, the flavonoids taxifolin and epigallocatechin-3-gallate (EGCG) could be explored as biomolecules in scaffolds for regenerative endodontic procedures. The aim of this study was to evaluate the effect of taxifolin and EGCG on the cell viability, differentiation, and expression of biomineralization markers in stem cells from the apical papilla. DESIGN: Stem cells from the apical papilla were exposed to single or continuous treatments with taxifolin at 200, 100 and 50 µM and EGCG at 50, 25 and 12.5 µM for 48 h, 8 and 14 days, in regular or mineralizing media. Cell viability, alkaline phosphatase activity and calcium deposition were analyzed using resazurin, p-nitrophenylphosphate and alizarin-based assays. RESULTS: None of the flavonoid groups affected cell viability at 48 h, however at 8 and 14 days, Taxifolin 200 µM and EGCG 50 µM were cytotoxic. Cells did not express alkaline phosphatase activity when grown in regular medium, even in the presence of flavonoids. Alkaline phosphatase activity and biomineralization potential were higher in cells treated with Taxifolin 50 µM and EGCG 12.5 µM. CONCLUSION: Taxifolin and EGCG exhibited a concentration, time, and therapeutic mode dependent bioactivity on stem cells from the apical papilla. Both flavonoids at the lower concentrations tested exhibited cytocompatibility and increased expression of mineralization markers in the presence of mineralizing agents.

Microbiological Properties and Cytotoxicity of PNVCL Hydrogels Containing Flavonoids as Intracanal Medication for Endodontic Therapy.

This study aimed to evaluate the cytotoxicity and microbiological properties of poly (N-vinylcaprolactam)-PNVCL hydrogels containing flavonoids as intracanal medication for endodontic therapy. Antimicrobial activity of ampelopsin (AMP), isoquercitrin and rutin was determined against Enterococcus faecalis, Actinomyces israelii, Lactobacillus casei, Streptococcus mutans, and Fusobacterium nucleatum by the microdilution method. After synthesis and characterization by rheology, PNVCL hydrogels were loaded with AMP and controls calcium hydroxide (CH) and chlorhexidine (CHX), and determined the compounds release profile. PNVCL+AMP, PNVCL+CH, PNVCL+CHX were evaluated on multi-species biofilms and analyzed by Scanning Electron Microscopy (SEM) and Confocal Laser Scanning Microscopy (CLSM). Cytotoxicity was determined after fibroblasts exposure to serial dilutions of AMP and PNVCL hydrogel extracts. AMP was effective against all of the bacteria tested, especially against S. mutans, A. israelli and F. nucleatum. SEM and CLSM analysis showed that PNVCL + AMP caused a significant decrease and disorganization of multi-species biofilms and reduction of intracanal viable cells, superior to the other groups. AMP affected fibroblast viability at concentrations above 0.125 mg/mL, and extracts of PNVCL+AMP showed low cytotoxicity. In conclusion, PNVCL containing AMP demonstrated cytocompatibility and potent effect against multi-species biofilms and could be potential intracanal medication for endodontic purposes.

Hardness of Resin Cements Polymerized through Glass-Ceramic Veneers.

(1) Background: The aim of this study is to evaluate the hardness of resin cements polymerized through ceramic disks under different process factors (ceramic type and thickness, light-polymerization units and polymerization time); (2) Method: Three types of ceramic blocks were used (IPS e.max CAD; Celtra Duo; VITABLOCS). Ceramic disks measuring 0.5 mm, 1.0 mm and 1.5 mm were cut from commercial blocks. Two resin cements (Rely X Veneer and Variolink Esthetic) were polymerized through the ceramic specimens using distinct light-polymerization units (Deep-cure; Blue-phase) and time intervals (10 and 20 s). Hardness of cement specimens was measured using microhardness tester with a Knoop indenter. Data were statistically analyzed using factorial ANOVA (α = 5%); (3) Results: Mean microhardness of Rely X Veneer cement was significantly higher than that of Variolink Esthetic. Deep-cure resulted in higher mean microhardness values compared to Blue-phase at 0.5- and 1-mm specimen thicknesses. Moreover, a direct correlation was found between polymerization time and hardness of resin cement; (4) Conclusions: Surface hardness was affected by resin cement type and ceramic thickness, and not affected by ceramic types, within evaluated conditions. Increasing light-polymerization time significantly increased the hardness of the cement.

Interaction of epigallocatechin-gallate and chlorhexidine with Streptococcus mutans stimulated odontoblast-like cells: Cytotoxicity, Interleukin-1ß and co-species proteomic analyses.

OBJECTIVES: The dentin therapeutic agent chlorhexidine has inflammatory and cytotoxic characteristics urging investigation of alternatives like the natural compound epigallocatechin-gallate. The aim is to verify the effect of epigallocatechin-gallate and chlorhexidine on viability, interleukin-1ß (IL-1ß) and differential protein expression of MDPC-23 odontoblast-like cells stimulated by Streptococcus mutans. DESIGN: Cells were stimulated with heat-killed S. mutans at multiplicity of infection (MOI) of 100-1000 and subsequently treated with 100-1 µM of epigallocatechin-gallate. Cells with no treatment or chlorhexidine were controls. Combined stimulated/treated cells were tested for cytotoxicity (Alamar-Blue, N = 3, n = 3), total protein (N = 3, n = 3), IL-1ß (ELISA, N = 3, n = 3), and differential protein expression by liquid chromatography-tandem mass spectrometry (LC-MS/MS, n = 2). RESULTS: Cells stimulated at MOI 100/1000 and treated with 10 µM epigallocatechin-gallate and chlorhexidine did not present cytotoxicity. IL-1ß significantly increased in both un-stimulated and stimulated chlorhexidine 10 µM groups when compared to un-treated control (p < 0.05). MOI 100 chlorhexidine 10 µM group significantly increased IL-1ß compared to un-stimulated chlorhexidine 10 µM and epigallocatechin-gallate 10 µM groups, as well as to MOI 100 epigallocatechin-gallate 10 µM group (p < 0.05). LC-MS/MS revealed S. mutans and mammalian proteins, with tooth-specific proteins exhibiting different abundance levels, depending on the tested condition. CONCLUSIONS: Odontoblast-like cells stimulated with S. mutans at different MOI combined with epigallocatechin-gallate treatment did not cause cytotoxicity. S. mutans stimulation combined with chlorhexidine 100 µM treatment decreased cell viability, while treatment with chlorhexidine 10 µM concentration significantly increased IL-1ß. S. mutans stimulation and treatment of cells resulted in varied protein expression.

Methacrylation of epigallocatechin-gallate for covalent attachment with a dental polymer.

OBJECTIVE: Synthesize novel epigallocatechin-gallate (EGCG) methacrylate monomers with the ability to copolymerize with dental methacrylate resins. METHODS: EGCG was reacted with 1/3 (E33), 2/3 (E67) and 1 (E100) molar equivalents of methacyloyl chloride introducing three degrees of polymerizablility. EGCG-methacrylates were characterized by Fourier Transform Infrared Spectroscopy (FTIR) and Nuclear Magnetic Resonance (NMR). E33, E67, E100 and neat EGCG were incorporated into TEGDMA at 0.5-20% ratios (m/m). Copolymers were tested for degree of conversion (%DC), EGCG release, gel content (%GC), degree of swelling (%DS), flexural properties and bacterial viability (Streptococcus mutans, baseline/30-days). Neat TEGDMA and TEGDMA passively loaded with EGCG (E0) were used as controls. Data were analysed by one-way ANOVA, Tukey, and Dunnett's method (α=5%). Two-way ANOVA and Bonferroni were used to investigate factor interaction. RESULTS: FTIR/NMR confirmed synthesis of desired compounds. All of E100 incorporated ratios had %DC similar to TEGDMA. Remaining groups had reduction in %DC at 2% in E0, 10% in E33 and 20% in E67 ratios. EGCG was stable within ECGC-methacrylate copolymers. Release of EGCG from E0 significantly increased with higher EGCG ratios. Except for E100, higher EGCG or EGCG-methacrylate ratios led to decreased %CG and %DS. At baseline, E0 had the lowest bacterial survival rates (1-10% survival) at all ratios compared to E33, E67, E100, and neat TEGDMA. However, E33, E67 and E100 still had statistically lower survival rates (7-53%) compared with neat TEGDMA. After 30-days, all compounds had similar survival rates for all ratios, which were lower than that of neat TEGDMA. SIGNIFICANCE: Demonstration of methacrylate functionalized EGCG- with inherited antibacterial activity for improved restoration longevity.

Experimental self-etching resin infiltrants on the treatment of simulated carious white spot lesions.

OBJECTIVES: To evaluate the penetration depth (µm) of experimental resin infiltrants containing different percentages of triethylene glycol dimethacrylate (TEGDMA) and phosphoric acid 2-hydroxyethyl methacrylate ester (PAM) in artificial carious white spot lesions (WSL). METHODS: WSL were produced in 65 bovine flat enamel specimens by pH cycling protocol, which were treated with either Icon (control) or experimental acidic infiltrants based on different percentages of TEGDMA and PAM monomers (acidic), and their association or not with previous acid-etching with phosphoric acid. Ten readings using Confocal Laser Scanning Microscopy were conducted on each specimen and the penetration depth was calculated from the surface until the deepest point with the fluorescent dye Rhodamine B (0.02 mg/mL). The pH and the viscosity of the experimental infiltrants were also tested. Data were statistically analyzed with two-way ANOVA and Tukey tests (α < 0.05). RESULTS: The material factor and the interaction material*acid-etching were statistically significant. The lowest penetration depth was observed for the samples treated with the commercial infiltrant after etching with 15% hydrochloric acid. When specimens were pre-treated with PA, highest penetration was seen for specimens treated with 100% TEGDMA, which differed from all other groups. The lowest penetration was seen for those treated with 50:50 TEGDMA:PAM infiltrants. When specimens were not previously etched, highest penetration was seen for Icon, which differed only from those treated with 25% TEGDMA 75% PAM, where the lowest values were seen. The values of viscosity increased and the pH decreased with the addition of PAM in the infiltrant formulations. CONCLUSION: the association between TEGDMA and PAM seems to allow similar infiltration depth reached by Icon infiltrant without acid etching the enamel surface.

Analysis of permeability and biological properties of dentin treated with experimental bioactive glasses.

OBJECTIVES: To evaluate obliterating capability and biological performance of desensitizing agents. METHODS: 50 dentin blocks were distributed according to the desensitizing agent used (n = 10): Control (Artificial saliva); Ultra EZ (Ultradent); Desensibilize Nano P (FGM); T5-OH Bioactive Glass (Experimental solution); F18 Bioactive Glass (Experimental solution). Desensitizing treatments were performed for 15 days. In addition, specimens were subjected to acid challenge to simulate oral environment demineralizing conditions. Samples were subjected to permeability analysis before and after desensitizing procedures and acid challenge. Cytotoxicity analysis was performed by using Alamar Blue assay and complemented by total protein quantification by Pierce Bicinchoninic Acid assay at 15 min, 24-h and 48-h time points. Scanning electron microscopy and energy dispersion X-ray spectroscopy were performed for qualitative analysis. Data of dentin permeability was analyzed by two-way repeated measures ANOVA and Tukey's test. For cytotoxicity, Kruskal-Wallis and Newman-Keuls tests. RESULTS: for dentin permeability there was no significant difference among desensitizing agents after treatment, but control group presented highest values (0.131 ± 0.076 Lp). After acid challenge, control group maintained highest values (0.044 ± 0.014 Lp) with significant difference to other groups, except for Desensibilize Nano P (0.037 ± 0.019 Lp). For cytotoxicity, there were no significant differences among groups. CONCLUSION: Bioglass-based desensitizers caused similar effects to commercially available products, regarding permeability and dentin biological properties. CLINICAL SIGNIFICANCE: There is no gold standard protocol for dentin sensitivity. The study of novel desensitizing agents that can obliterate dentinal tubules in a faster-acting and long-lasting way may help meet this clinical need.

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.

In situ effect of a proanthocyanidin mouthrinse on dentin subjected to erosion.

BACKGROUND: Proanthocyanidin has been shown to be efficient in inhibiting matrix metalloproteinases. OBJECTIVE: The aim of this in situ study was to evaluate the protective effect of Proanthocyanidin-based mouthrinses either with naturally acidic or with a neutral pH applied on dentin subjected to erosion. METHODOLOGY: Eight volunteers wore one palatal device in two phases (7 days washout) with 16 samples per group (n=8). The groups under study were: First Phase/ G1 - 10% proanthocyanidin mouthrinse (pH 7.0, Experimental group 1 - Purified Grape Seeds Oligomeric Proanthocyanidins), G2 - 10% proanthocyanidin mouthrinse (pH 3.0, Experimental group 2 - Purified Grape Seeds Oligomeric Proanthocyanidins). Second Phase/ G3 - 0.12% chlorhexidine mouthrinse (pH 7.0, Positive control group), G4 - no previous treatment (Negative control group). Each device was subjected to 3 erosive cycles (5 minutes) per day for 5 days. Treatments with different mouthrinses were applied once after the second erosive challenge (5 minutes). Profilometry was used to quantify dentin loss (µm). RESULTS: Data were analyzed by repeated measures of ANOVA followed by Fisher's test (p<0.05). G1 (1.17±0.69) and G3 (1.22±0.25) showed significantly lower wear values with no statistical difference between them. G2 (2.99±1.15) and G4 (2.29±1.13) presented higher wear values with no significant differences between them. CONCLUSION: The 10% proanthocyanidin mouthrinse (pH 7.0) could be a good strategy to reduce dentin wear progression.

Influence of glutaraldehyde priming on bond strength of an experimental adhesive system applied to wet and dry dentine.

OBJECTIVES: This study tested the following null hypotheses: (1) there is no difference in resin-dentine bond strength when an experimental glutaraldehyde primer solution is added prior to bonding procedures and (2) there is no difference in resin-dentine bond strength when experimental glutaraldehyde/adhesive system is applied under dry or wet demineralized dentine conditions. METHODS: Extracted human maxillary third molars were selected. Flat, mid-coronal dentine was exposed for bonding and four groups were formed. Two groups were designated for the dry and two for the wet dentine technique: DRY: (1) Group GD: acid etching+glutaraldehyde primer (primer A)+HEMA/ethanol primer (primer B)-under dried dentine+unfilled resin; (2) Group D: the same as GD, except for primer A application; WET: (3) Group GW: the same as GD, but primer B was applied under wet dentine condition; (4) Group W: the same as GW, except for primer A application. The bonding resin was light-cured and a resin core was built up on the adhesive layer. Teeth were then prepared for microtensile bond testing to evaluate bond strength. The data obtained were submitted to ANOVA and Tukey's test (alpha=0.05). RESULTS: Glutaraldehyde primer application significantly improved resin-dentine bond strength. No significant difference was observed when the same experimental adhesive system was applied under either dry or wet dentine conditions. These results allow the first null hypothesis to be rejected and the second to be accepted. CONCLUSION: Glutaraldehyde may affect demineralized dentine properties leading to improved resin bonding to wet and dry substrates.