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.

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

Abstract 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.

PREVENTIVE EFFECT OF CRANBERRY GEL ON DENTIN SUBMITTED TO EROSION

Aims: Considering that Cranberry's components might inhibit dentin metalloproteinases exposed to erosive agents, the aim of this study was to evaluate in situeffect of a Cranberry gel application on dentin before an erosive challenge.Materials and methods: This crossover double-blinded study was performed in 2 phases of 5 days each, with 10 healthy volunteers who wore 2 palatal devices (1 for each phase) with 4 dentin specimens (2 specimens for each group). The groups under study were:First Phase: G1 -Erosivechallenge (Coca-cola®) over dentin without any previous treatment (1st negative control group); G2 -Erosive challenge over dentin previously treated with Cranberry gel (test group); and Second Phase: G3 -Erosive challenge over dentin previously treated with a gel without any active principle (2ndnegative control group); G4 -Erosive challenge over dentin previously treated with 0.12% Chlorhexidine gel (positive control group). Each device was immersed into the acid beverage, 3 times daily for 5 minutes during 5 days. Profilometry (µm) was used to quantify the dentin wear. Data were analyzed by Repeated Measures Analysis of Variance followed by Fisher's test (p<0.05).Results: Data (G1: 4.98 ± 1.36a; G2: 3.29 ± 1.16b; G3: 4.38 ± 1.19a; G4: 3.32 ± 1.55b) showed no statistical difference between G1 and G3. There was also no difference between G2 and G4. However, G2 and G4 presented lower wear when compared to G1 and G3, and this difference was statistically significant. Conclusion: The results of this studysuggest a significant efficacy of Cranberry gel in preventing wear of dentin subjected to dental erosion.

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.

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.

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.

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.

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

Abstract 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.

Insights on the role of cytokines in carious lesions / Insights sobre o papel das citocinas em lesões cariosas

Objetivo: A resposta imune da dentina-polpa à patogênese da cárie ainda é pouco compreendida devido à complexa interação dos processos envolvidos. O objetivo desta revisão foi explorar o papel das citocinas e sua relevância na patogênese da cárie dental. Resultados: A cárie dentária pode resultar em uma resposta inflamatória do hospedeiro na polpa dental, caracterizada pelo acúmulo de células inflamatórias levando à liberação de citocinas inflamatórias como, Interleucina-4 (IL-4), Interleucina (IL-6), Interleucina-8 (IL-8) e fator de necrose tumoral­α(TNF-α). IL-4 parece estar correlacionada com a profundidade das lesões cariosas; IL-6 está fortemente correlacionada com a doença cárie e é considerada um potente biomarcador; IL-8 pode ser um potente biomarcador tanto para cárie quanto para outras alterações presentes na polpa e sua liberação está correlacionada com TNF-α e IL-6; TNF-α desempenha um papel importante não apenas na progressão da cárie, mas também em outros processos patológicos. Conclusao: Mediadores específicos têm um grande potencial para servir como biomarcadores quanto à presença e progressão da doença cárie, o que incita a necessidade de mais investigações nesse campo (AU).

Objectives: The dentin-pulp immune response to caries pathogenesis is still poorly understood due to the complex interplay of the involving processes. The aim of this review was to explore the role of cytokines and its relevance in the pathogenesis of dental caries. Results: Dental caries can result in a host inflammatory response in the dental pulp, characterized by the accumulation of inflammatory cells leading to the release of inflammatory cytokines such as Interleukin-4 (IL-4), Interleukin (IL-6), Interleukin-8 (IL-8) and Tumor necrosis factor­ α (TNF- α ). IL-4 seems to be correlated to the depth of carious lesions; IL-6 is strongly correlated to caries disease and is considered a potent biomarker; IL-8 can be a potent biomarker for both caries and other changes present in the pulp and, its release is correlated to TNF- α and IL-6; TNF-α plays an important role not only in caries progression, but also in other pathological processes. Conclusion: Specific mediators have a great potential to serve as biomarkers alluding to the presence and progress of caries disease, urging further investigations in the field (AU)

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.

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.

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).

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.

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.

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.

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.

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.

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.

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.