Role of indoleamine 2,3-dioxygenase in an inflammatory model of murine gingiva.

BACKGROUND AND OBJECTIVE: Indoleamine 2,3-dioxygenase (IDO) is one of the major pathways for metabolism of tryptophan in a variety of cells, including immune cells. Increasing evidence indicates that IDO is a critical player in establishing the balance between immunity and tolerance and ultimately in the maintenance of homeostasis. By inducing inflammation in gingival tissue, we tested the hypothesis that IDO is a pivotal player in regulating the immune and inflammatory responses of gingiva. MATERIAL AND METHODS: We utilized the IDO knockout mouse model in conjunction with lipopolysaccharide (LPS)-induced inflammation. Accordingly, wild-type and IDO knockout mice were injected with LPS or vehicle in the anterior mandibular gingiva, twice over a 2-wk period, which was followed by procurement of gingival tissue for histopathology and preparation of tissue for flow cytometry-based studies. RESULTS: Clinical and histological examinations revealed a marked adverse impact of IDO deficiency on gingival inflammation. These observations were consistent with a more marked increase in the number of cells positive for the proinflammatory cytokine interleukin (IL)-17, but no significant change in the number of cells positive for the anti-inflammatory cytokine IL-10, in LPS-treated IDO knockout mice. Consistent with the more marked proinflammatory impact of IDO deficiency, the percentage of regulatory T cells was much reduced in gingival tissue of LPS-treated IDO knockout mice than in gingival tissue of wild-type mice. These proinflammatory changes were accompanied with a prominent increase in apoptotic and necrotic cell death in gingival tissue of IDO knockout mice compared with wild-type mice. CONCLUSION: Collectively, our findings support a major role for IDO in the development of gingival inflammation, as an example of an inflammatory condition, and lay the foundation for subsequent studies to explore it as a novel immunotherapy target.

Effect of air-drying on the solvent evaporation, degree of conversion and water sorption/solubility of dental adhesive models.

This study evaluated protocols to eliminate acetone from dental adhesives and their effect on the kinetic of water sorption and percent of conversion of these adhesives. Experimental methacrylate-based adhesives with increasing hydrophilicity (R2, R3, R5) were used as reference materials. Primer-like solutions were prepared by addition of 50 wt% acetone. Acetone elimination was measured gravimetrically before and after: a spontaneous evaporation, an application of air-drying at room temperature or application of 40°C air-drying. Protocols were performed from 15 to 60 s. Specimens of adhesive/acetone mixtures were photo-activated and tested for degree of conversion, water sorption and solubility. Data were analyzed by ANOVA and Bonferroni's tests (α = 0.05). Complete acetone elimination was never achieved, but it was significantly greater after the 40°C air-drying application. Higher acetone elimination was observed for the least hydrophilic adhesive. Longer periods for acetone evaporation and heated air-stream can optimize polymerization and reduce the water sorption/solubility of adhesive system models.

Mechanical Stability and Proteolytic Activity of Resin-dentin Bonds Using the Cross-linked Dry Bonding Technique.

OBJECTIVE: To evaluate the mechanical stability and the proteolytic activity of bonds created by a two-step, etch-and-rinse adhesive applied to cross-linked and air-dried etched dentin. METHODS: Flat dentin surfaces were produced in 64 extracted sound human molars. The dentin was etched with 35% phosphoric acid for 15 seconds, and then the teeth were divided into groups according to the cross-linking solution applied on the etched dentin. Group 1: 5% grape seed extract (GSE), Group 2: 5% glutaraldehyde, Group 3: Gluma Desensitizer, or Group 4: deionized water (control). Solutions were applied for 60 seconds, followed by rinse and blot drying. Then, the teeth were separated into two subgroups where the etched dentin was kept moist or air-dried. The adhesive was applied followed by a composite resin buildup. After 24 hours, the teeth were cut into beams (0.81 mm²) that were tested for microtensile strength immediately or after 12 months of aging in a 37°C saliva-like buffer. Additional teeth (n=32) were bonded as described and cut into 0.5-mm-thick slabs. The slabs were prepared for nanoleakage (scanning electron microscopy) and in situ zymography (EnzChek Protease Assay Kit). Bond strength data were submitted to ANOVA and Tukey tests (α =0.05). RESULTS: Significant reduction in immediate bond strength (ca 65%) and increase in proteolytic activity was seen when the etched dentin was air dried without previous cross-linking biomodification. Conversely, bond strengths did not differ from those produced on wet dentin when collagen was cross-linked before air drying, irrespective of the solution applied. For both moist and air-dried etched dentin, collagen cross-linking resulted in mechanically stable bonds and reduced proteolytic activity after 12 months of storage. CONCLUSION: Bonds produced by the application of a two-step, etch-and-rinse adhesive to cross-linked, air-dried, etched dentin were mechanically stable and revealed reduced proteolytic activity after 1 year of aging.

Canal and isthmus debridement efficacies of two irrigant agitation techniques in a closed system.

AIM: To compare canal and isthmus debris debridement efficacies of the manual dynamic irrigation (MDI) and apical negative pressure (ANP) techniques in the mesial root of mandibular first molars with narrow isthmi, using a closed canal design. METHODOLOGY: Micro-computed tomography was employed to select 20 teeth, each containing a narrow isthmus. Each root was sealed at the apex with hot glue and embedded in polyvinylsiloxane to simulate a closed canal system. The teeth were submitted to a standardized instrumentation protocol. Final irrigation was performed with either the MDI or the ANP technique using the EndoVac system (N=10). Masson trichrome-stained sections were prepared from completely demineralized roots at 10 canal levels between 1 and 2.8mm of the anatomical apices. Areas occupied by canals and isthmus of each root and debris in the corresponding regions were digitized by the NIH Image J software and statistically analysed using two-way repeated measures anova. RESULTS: For the instrumented canals, there were no differences between the two groups (P=0.131) in the area occupied by debris at all canal levels (P=0.343). Conversely, for the isthmus, less debris was found in the ANP group (P<0.001) but no differences were seen in each group with respect to the 10 canal levels (P=0.352). CONCLUSION: Neither technique completely removed debris from the isthmus regions. However, the EndoVac system, which encompasses the ANP concept, removed considerably more debris from narrow isthmi in mandibular mesial roots.

Root canal debridement using manual dynamic agitation or the EndoVac for final irrigation in a closed system and an open system.

AIM: This study examined canal debridement efficacy by testing the null hypothesis that there is no difference between a 'Closed' and an 'Open' system design in smear layer and debris removal using either manual dynamic agitation or the EndoVac for irrigant delivery. METHODOLOGY: Forty teeth were divided into four groups and submitted to a standardized instrumentation protocol. Final irrigation was performed with either manual dynamic agitation or the EndoVac on groups of teeth with or without a sealed apical foramen. Smear and debris scores were evaluated using SEM and analysed using Cochran-Mantel-Haenszel statistic. RESULTS: The ability of manual dynamic agitation to remove smear layer and debris in a closed canal system was significantly less effective than in an open canal system and significantly less effective than the EndoVac (P<0.001). CONCLUSION: The null hypothesis was rejected; the presence of a sealed apical foramen adversely affected debridement efficacy when using manual dynamic agitation but not the EndoVac. Apical negative pressure irrigation is an effective method to overcome the fluid dynamics challenges inherent in closed canal systems.

The effect of ultrasonic removal of various root-end filling materials.

AIM: To compare residual root-end filling material in apical root-end cavities following their removal with ultrasonic retrotips. METHODOLOGY: Thirty single-rooted teeth were filled with Thermafil and AH Plus sealer. Root-ends were resected at 90 degrees, 3 mm from the apex. Root-end cavities were prepared with diamond burs and ultrasonic retrotips and filled with one of three filling materials: group I: Retro-TC (calcium silicate-based cement), group II: IRM (Dentsply, Germany), group III: Vitrebond (3M ESPE, USA). After 30 days of storage, ultrasonic retrotips were used to remove materials from the root-end cavities. The ultrasonic application time was fixed at 60 s. Polyether impressions and replicas of the root-ends were made. Root apices and replicas were examined by one operator under a scanning electron microscope. Remnants of residual materials were evaluated using a four-level scoring system; fractures, smear layer and exposed dentinal tubules were also examined. RESULTS: Forty per cent of the specimens filled with Retro-TC revealed complete removal of the material with exposure of dentinal tubules, whilst 60% contained residual cement. Twenty per cent of specimens filled with IRM were completely devoid of material, whereas 80% had retained material. Ten per cent of specimens filled with Vitrebond retained a moderate amount of material whilst 90% had substantial retention of the material. Statistically significant differences were found (P < 0.05) amongst the three groups of materials. CONCLUSIONS: Retro-TC was successfully removed in 40% of cases using ultrasonics retrotips for 60 s, whereas IRM and Vitrebond specimens had evidence of retained material in 80% and 90% of all specimens respectively.

Dislocation resistance of ProRoot Endo Sealer, a calcium silicate-based root canal sealer, from radicular dentine.

AIM: To examine the dislocation resistance of three root canal sealers from radicular dentine with and without immersion in a simulated body fluid (SBF), using a modified push-out test design that produced simulated canal spaces of uniform dimensions under identical cleaning and shaping conditions. METHODOLOGY: Sixty single-rooted caries-free human canine teeth were used. Standardized simulated canal spaces were created using 0.04 taper ProFile instruments along the coronal, middle and apical thirds of longitudinal tooth slabs. Following NaOCl/ethylenediamine tetra-acetic acid cleaning, the cavities were filled with ProRoot Endo Sealer, AH Plus Jet or Pulp Canal Sealer. After setting, half of the cavities were tested with a fibre-optic light-illuminated push-out testing device. The rest were immersed in SBF for 4 weeks before push-out evaluation. Failure modes were examined with stereomicroscopy and field emission (FE)-scanning electron microscopy. RESULTS: Location of the sealer-filled cavities did not affect push-out strengths. ProRoot Endo Sealer exhibited higher push-out strengths than the other two sealers particularly after SBF storage (P < 0.001). Failure modes were predominantly adhesive and mixed for Pulp Canal Sealer and AH Plus Jet, and predominantly cohesive for ProRoot Endo Sealer. Spherical amorphous calcium phosphate-like phases that spontaneously transformed into apatite-like phases were seen in the fractured specimens of ProRoot Endo Sealer after SBF storage. CONCLUSIONS: When tested in bulk without a main core, both 'sealer type' and 'SBF storage' were significant in affecting push-out results. The ProRoot Endo Sealer demonstrated the presence of spherical amorphous calcium phosphate-like phases and apatite-like phases (i.e. ex vivo bioactivity) after SBF storage.

In vivo chlorhexidine stabilization of hybrid layers of an acetone-based dentin adhesive.

The current in vivo study evaluated the degradation of dentin hybrid layers in deep occlusal-surface resin composite restorations using TEM. Caries-free premolars scheduled for extraction as part of orthodontic treatment were prepared and restored, then extracted after 12 months. The adhesive used was a single-bottle etch-and-rinse acetone-based product (Prime & Bond NT, Dentsply/Caulk). Control group restorations (n=8) were placed according to the manufacturer's instructions, while the experimental group received application of a 2% solution of chlorhexidine digluconate after etching and rinsing and prior to application of the adhesive. Extensive degradation was observed in all of the teeth in the control group after 12 months, while no degradation was observed in the experimental group. In vitro testing showed no significant difference in immediate microtensile bond strength between the control and experimental adhesive protocols.

Chitosan-Based Extrafibrillar Demineralization for Dentin Bonding.

Instability of resin-dentin bonds is the Achilles' heel of adhesive dentistry. To address this problem, a chelate-and-rinse extrafibrillar dentin demineralization strategy has been developed that keeps intrafibrillar minerals within collagen fibrils intact to prevent activation of endogenous proteases that are responsible for collagen degradation within hybrid layers. The objective of the present study was to evaluate the potential of using chitosan >40 kDa as an antimicrobial extrafibrillar dentin-chelating agent to enhance bond durability. Transmission electron microscopy provided evidence for retention of intrafibrillar minerals and smear plugs in dentin conditioned with 1 wt% chitosan. Analyzed by Kruskal-Wallis analysis of variance, Dunn's statistic, and separate Mann-Whitney tests, tensile bond strengths to wet- and dry-bonded dentin indicated that chelating dentin with chitosan for 60 s prior to bonding did not result in a significant decline in resin-dentin bond strength when compared with that of phosphoric acid etching ( P > 0.05). Gelatinolytic activity within the hybrid layers was examined via in situ zymography after 24-h storage or after thermomechanical cycling and analyzed with 3-factor analysis of variance. After 24 h, enzymatic activity was detected only within completely demineralized phosphoric acid-etched dentin, with values derived from dry bonding significantly higher than those derived from wet bonding ( P < 0.05). Negligible fluorescence was detected within hybrid layers when dentin was conditioned with chitosan, even after thermomechanical cycling, as compared with the controls. Reduction in water permeability in chitosan-conditioned dentin, attributed to smear plug retention, also fostered long-term bond stability. Antibacterial testing performed with live/dead staining indicated that the acetic acid-solubilized chitosan possessed antibacterial activities against 3 single-species biofilms: Streptococcus mutans, Actinomyces naeslundii, and Enterococcus faecalis. Taken together, the new chitosan-based extrafibrillar demineralization strategy retains intrafibrillar minerals, reduces endogenous protease-initiated collagen degradation, prevents water permeation within hybrid layers, and kills bacteria on dentin surfaces, which are crucial factors for enhancing resin-dentin bond durability.

Changes in resin-infiltrated dentin stiffness after water storage.

Plasticization of polymers by water sorption lowers their mechanical properties in a manner that is predictable by the polarity of their component resins. This study tested the hypothesis that when adhesive resins were used to create resin-infiltrated dentin, the reductions in their flexural moduli after water storage would be lowered proportional to their hydrophilic characteristics. Three increasingly hydrophilic resin blends were used to fabricate polymer beams and macro-hybrid layer models of resin-infiltrated dentin for testing with a miniature three-point flexure device, before and after 1-4 weeks of water storage. Flexural modulus reductions in macro-hybrid layers were related to, and more extensive than, reductions in the corresponding polymer beams. Macro-hybrid layers that were more hydrophilic exhibited higher percent reductions in flexural modulus, with the rate of reduction proportional to the Hoy's solubility parameters for total intermolecular attraction forces (delta(t)) and polar forces (delta(p)) of the macro-hybrid layers.

Chlorhexidine release and water sorption characteristics of chlorhexidine-incorporated hydrophobic/hydrophilic resins.

OBJECTIVES: The aim of this study was to evaluate chlorhexidine release from unfilled non-solvated methacrylate-based resins of increasing hydrophilicity and to examine relationships among Hoy's solubility parameters, water sorption, solubility and the rate of chlorhexidine release. METHODS: Resin discs were prepared from light-cured, experimental resin blends (R1, R2, R3, R4 and R5) containing 0.0, 0.2, 1.0 and 2.0 wt.% chlorhexidine diacetate (CDA). Discs were immersed in distilled water at 37 degrees C, and mass changes were recorded at different periods. Spectral measurements were made to follow change in optical densities of storage solution to examine chlorhexidine release kinetics. After a 28-day period, water sorption, solubility, and the cumulative chlorhexidine release were obtained. Additionally, antibacterial study was performed by observing the presence of inhibition zone against Streptococcus mutans. RESULTS: The most hydrophilic resin (R5) exhibited the highest chlorhexidine release rate. The most hydrophobic resin (R1) exhibited the lowest rate. However, no inhibition zone was produced by any specimens stored in water for 2 weeks. The addition of CDA increased solubility significantly but had no effect on water sorption. Significant positive correlations were seen between water sorption and the cumulative chlorhexidine release. SIGNIFICANCE: Chlorhexidine release from resins may be related to water-induced swelling, which in turn is enhanced by the hydrophilicity of cured polymer matrix.

Microscopic appearance and apical seal of root canals filled with gutta-percha and ProRoot Endo Sealer after immersion in a phosphate-containing fluid.

AIM: To investigate the sealing quality of ProRoot Endo Sealer, a calcium silicate-based sealer and its morphologic characteristics after immersion in a phosphate-containing fluid (PCF). METHODOLOGY: Single-rooted canals were filled with gutta-percha and either ProRoot Endo Sealer or two commercially available zinc oxide eugenol (ZOE)-based and epoxy resin-based sealers. The sealers were allowed to set for 6 days and the filled teeth were immersed in PCF for 24 h before fluid leakage evaluation. After initial leakage evaluation at the 7th day, each filled root was restored and reimmersed in PCF for 28 days before the second phase of leakage evaluation at 35 days. Cryofractured specimens of additional teeth filled with the three sealers were examined using scanning electron microscopy after immersion in PCF for the two periods. RESULTS: One-way repeated measures anova and Tukey test revealed significant differences between the ZOE-based sealer at 35 days and the calcium silicate-based sealer at 35 days (P < 0.001), and between the ZOE-based sealer at 7 days and the calcium silicate-based sealer at 35 days (P = 0.001). No difference was found between the epoxy resin-based sealer and the calcium silicate-based sealer after both storage periods. Cryofractured calcium silicate-based sealer specimens demonstrated apatite-like crystalline deposits along the apical and middle thirds of the canal walls via transformation from amorphous calcium phosphate-like precursors. CONCLUSIONS: ProRoot Endo Sealer is comparable in sealing quality to the epoxy resin-based sealer and seals better than the ZOE-based sealer after immersion in PCF. The calcium silicate-based sealer also demonstrates ex vivo bioactivity when it comes into contact with phosphate ions.

Discovering new treatments for sensitive teeth: the long path from biology to therapy.

Tooth sensitivity is a common dental pain condition where sufferers experience brief episodes of sharp well-localized pain when their teeth are subjected innocuous stimuli such as cold, air-currents and probing with a metallic instrument. In this review, we will make no attempt to describe all the treatments that have been developed to treat tooth sensitivity. We will review the basic anatomic and physiological mechanisms responsible for sensitivity. The insights into the dental lesions responsible for tooth sensitivity, as well as the physiological processes linking stimuli and pain generation have suggested several treatments and preventive strategies. Unfortunately, many tooth sensitivity treatments fail to perform better than placebos in clinical trials that seek to assess the effect of agents on pain symptoms. In the case of the most commonly used self-applied desensitizing agent, potassium salts, the mechanism of action established by laboratory and animal models may not apply to clinical use. Thus results obtained with laboratory and animal models must be applied with care to clinical use. Clinical literature suggests that tooth sensitivity is the symptomatic manifestation of significant dental problems, such as wear and other forms of non-carious tooth structure loss. These conditions are increasing in frequency as people age, retaining their natural teeth longer. They are frequently the consequences of aggressive oral hygiene practices and diets rich in acids. Treatments directed at the underlying causes rather than the symptoms of tooth sensitivity would hinder the development of these lesions and provide researchers with objective targets for assessing therapeutic efficacy.

Zymography of Hybrid Layers Created Using Extrafibrillar Demineralization.

A chelate-and-rinse extrafibrillar calcium chelation dentin bonding concept has recently been developed and investigated for its effectiveness in improving resin-dentin bonding by bridging the gap between wet and dry dentin bonding. The objective of the present study was to evaluate the gelatinolytic activity of hybrid layers (HLs) created using the chelate-and-rinse bonding technique. Gelatinolytic activity within the HL was examined using in situ zymography and confocal laser-scanning microscopy after 24-h storage or after thermomechanical cycling. Dentin specimens were bonded with Prime&Bond NT (Dentsply Sirona) after conditioning with 15 wt% phosphoric acid for 15 s (control) or 15 wt% polymeric chelators (sodium salt of polyacrylic acid; PAAN) of 2 different molecular weights for 60 s. For each reagent, bonding was performed using dry-bonding and wet-bonding techniques ( n = 10). Slices containing the adhesive-dentin interface were covered with fluorescein-conjugated gelatin and examined with a confocal laser-scanning microscope. Fluorescence intensity emitted by the hydrolyzed fluorescein-conjugated gelatin was quantified. Gelatinolytic activity was expressed as the percentage of green fluorescence emitted within the HL. After storage for 24 h, enzymatic activity was only detected within the completely demineralized phosphoric acid-etched dentin, with values derived from dry bonding higher than those from wet bonding ( P < 0.05). Almost no fluorescence signals were detected within the HL when dentin was conditioned with PAANs compared with the controls ( P < 0.05). After thermomechanical cycling, enzymatic activities significantly increased for the phosphoric acid-conditioned, drying-bonding group compared with 24-h storage ( P < 0.05). The present study showed that the use of the chelate-and-rinse bonding concept for both dry-bonding and wet-bonding approaches results in the near absence of matrix-bound collagenolytic activities in the HL even after aging. This may be attributed to fossilization of endogenous proteases via preservation of intrafibrillar minerals within the dentin collagen matrix.

Tubular occlusion optimizes bonding of hydrophobic resins to dentin.

Although hydrophobic resins may be bonded to acid-etched dentin with an ethanol wet-bonding technique, the protocol is sensitive to moisture contamination when bonding is performed in deep dentin. This study tested the hypothesis that the use of oxalate or poly(glutamic) acid-modified, diluted ceramicrete (PADC) for dentinal tubule occlusion prevents fluid contamination and improves the bonding of an experimental hydrophobic adhesive to acid-etched, ethanol-dehydrated dentin. Mid-coronal and deep acid-etched moist dentin pre-treated with oxalate or PADC was dehydrated by ethanol wet-bonding and infiltrated with the experimental three-step etch-and-rinse hydrophobic adhesive under simulated pulpal pressure. Tensile bond strengths to deep dentin without pre-treatment were severely compromised. Conversely, oxalate and PADC pre-treatments reduced dentin permeability, prevented water contamination, and improved bond strengths. Minimal nanoleakage was identified within hybrid layers created in the oxalate- and PADC-pre-treated deep dentin. The use of tubular occluding agents optimized bonding of hydrophobic resins to dentin.

Bonding BisGMA to dentin--a proof of concept for hydrophobic dentin bonding.

The use of TEGDMA as a diluent comonomer in the formulation of hydrophobic adhesives for ethanol wet-bonding is a concern, due to its leaching potential, higher water sorption, and bio-incompatibility. This study tested the hypothesis that hydrophobic bonding to acid-etched dentin may be accomplished with the use of ethanol-solvated BisGMA only. Phosphoric-acid-etched, oxalate-occluded, deep coronal dentin bonded under 20 cm water pressure with experimental BisGMA adhesives by ethanol wet-bonding exhibited tensile strengths that were not significantly different from that achieved with OptiBond FL bonded according to the manufacturer-recommended protocol, with similar acid-/base-resistant hybrid layers, resin tags, and nanoleakage distribution. Ethanol replacement of water-saturated dentin produced wider interfibrillar spaces, more extensive shrinkage of the collagen fibrils, and narrower hybrid layers. Experimental BisGMA adhesives provide the proof of concept that relatively hydrophobic resins may be coupled to acid-etched dentin by increasing its hydrophobic characteristics via ethanol replacement. They should be further optimized before clinical application.

Chlorhexidine preserves dentin bond in vitro.

Loss of hybrid layer integrity compromises resin-dentin bond stability. Matrix metalloproteinases (MMPs) may be partially responsible for hybrid layer degradation. Since chlorhexidine inhibits MMPs, we hypothesized that chlorhexidine would decelerate the loss of resin-dentin bonds. Class I preparations in extracted third molars were sectioned into two halves. One half was customarily restored (etch-and-rinse adhesive/resin composite), and the other was treated with 2% chlorhexidine after being acid-etched before restoration. Specimens were stored in artificial saliva with/without protease inhibitors. Microtensile bond strengths and failure mode distribution under SEM were analyzed immediately after specimens' preparation and 6 months later. With chlorhexidine, significantly better preservation of bond strength was observed after 6 months; protease inhibitors in the storage medium had no effect. Failure analysis showed significantly less failure in the hybrid layer with chlorhexidine, compared with controls after 6 months. In conclusion, this in vitro study suggests that chlorhexidine might be useful for the preservation of dentin bond strength.

Zymographic analysis and characterization of MMP-2 and -9 forms in human sound dentin.

The role and function of dentin matrix metalloproteinases (MMPs) are not well-understood, but they may play a key role in dentinal caries and the degradation of resin-bonded dentin matrices. To test the null hypothesis that MMP-9 is not found in dentin matrix, we used gelatin zymography to extract and isolate all molecular forms of gelatinolytic MMPs in demineralized mature sound dentin powder obtained from extracted human molars, characterizing and identifying the enzymes by Western blotting. Gelatinolytic MMPs were detected in extracts of demineralized dentin matrix and identified as MMP-2 and MMP-9. Acidic extracts (pH 2.3) yielded 3-8 times more MMP activity than did EDTA (pH 7.4). Their activation may contribute to dentin matrix degradation, which occurs during caries progression and following resin bonding. Inhibition of MMP-2 and -9 proteolytic activity may slow caries progression and increase the durability of resin-dentin bonds.

Effect of dimethyl sulfoxide on dentin collagen.

OBJECTIVES: Infiltration of adhesive on dentin matrix depends on interaction of surface and adhesive. Interaction depends on dentin wettability, which can be enhanced either by increasing dentin surface energy or lowering the surface energy of adhesive. The objective was to examine the effect of dimethyl sulfoxide (DMSO) on demineralized dentin wettability and dentin organic matrix expansion. METHODS: Acid-etched human dentin was used for sessile drop contact angle measurement to test surface wetting on 1-5% DMSO-treated demineralized dentin surface, and linear variable differential transformer (LVDT) to measure expansion/shrinkage of dentinal matrix. DMSO-water binary liquids were examined for surface tension changes through concentrations from 0 to 100% DMSO. Kruskal-Wallis and Mann-Whitney tests were used to test the differences in dentin wettability, expansion and shrinkage, and Spearman test to test the correlation between DMSO concentration and water surface tension. The level of significance was p<0.05. RESULTS: Pretreatment with 1-5% DMSO caused statistically significant concentration-dependent increase in wetting: the immediate contact angles decreased by 11.8% and 46.6% and 60s contact angles by 9.5% and 47.4% with 1% and 5% DMSO, respectively. DMSO-water mixtures concentration-dependently expanded demineralized dentin samples less than pure water, except with high (≥80%) DMSO concentrations which expanded demineralized dentin more than water. Drying times of LVDT samples increased significantly with the use of DMSO. SIGNIFICANCE: Increased dentin wettability may explain the previously demonstrated increase in adhesive penetration with DMSO-treated dentin, and together with the expansion of collagen matrix after drying may also explain previously observed increase in dentin adhesive bonding.

Substantivity of Carbodiimide Inhibition on Dentinal Enzyme Activity over Time.

The use of 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide HCl (EDC) has recently been investigated for its effectiveness in the prevention of collagen degradation over time and the improvement of resin-dentin bond durability. The objective of the present study was to evaluate the effects of a 0.3 M EDC-containing conditioner on endogenous enzymatic activities within the hybrid layer (HL) created by a self-etch or an etch-and-rinse adhesive after 1 y. The activity within the HL was examined using in situ zymography and confocal laser scanning microscopy after 24 h or 1-y storage in artificial saliva. Dentin specimens were bonded with Clearfil SE Bond (CSE) or XP Bond (XPB). For CSE, the self-etching primer was applied and treated with 0.3 M EDC for 1 min, and then the bonding agent was applied. For XPB, dentin was etched and treated with 0.3 M EDC for 1 min and then bonded with the primer-bonding agent. Control specimens were prepared without EDC treatment. Slices containing the adhesive-dentin interface were covered with fluorescein-conjugated gelatin and observed with a multiphoton confocal microscope. Fluorescence intensity emitted by hydrolyzed fluorescein-conjugated gelatin was quantified, and the amount of gelatinolytic activity was represented by the percentage of green fluorescence emitted within the HL. After 24 h of storage, enzymatic activity was detected by in situ zymography within the HLs of both tested adhesives, with XPB higher than CSE ( P < 0.05). Almost no fluorescence signal was detected when specimens were pretreated with EDC compared to controls ( P < 0.05). After 1 y of storage, enzymatic activities significantly increased for all groups (excluding XPB control) compared to 24-h storage ( P < 0.05), with EDC pretreated specimens exhibiting significantly lower activity than controls ( P < 0.05). The present study showed, for the first time, that the use of EDC for both the self-etch and the etch-and-rinse approaches results in the reduction but not complete inhibition of matrix-bound collagenolytic enzyme activities over time in the HL.