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In vitro quantification of changes in human dentine tubule parameters using SEM and digital analysis.

Dentine hypersensitivity is recognized as a pain arising from fluid movement within dentine tubules that are open to the oral environment. Blocking the tubules is considered to be the principal aim of treatment, and the accurate assessment of tubule occlusion is the primary goal of many in vitro studies. This assessment usually comprises either measuring tubule permeability or scanning electron microscope examination of the dentine surface. Several scanning microscopy studies have claimed to quantify tubule occlusion, but are descriptive, qualitative or semi-quantitative evaluations. The present study was undertaken to assess the use of digital image analysis in quantifying the effectiveness of a selected desensitizing agent from micrographs of control and treated dentine surfaces. Using a dentine disc model, an accurate methodology was sought to investigate the occluding potential of Butler Protect (J.O. Butler, Chicago, IL, USA). Subjective examination of the images indicated there was little difference after a single application, but considerable effect after 20 applications. Quantitative digital analysis of a test image, demonstrated reproducibility between two examiners when used in fully- and semi-automated mode. After a single application of Butler Protect, multilevel statistical modelling demonstrated decreases in tubule area and maximum, minimum and mean diameter measurements (P < 0.001), whereas single level analysis showed increases in area and maximum and mean diameters. Multiple application of Butler Protect demonstrated even greater decreases in all parameters (P < 0.001). This quantitative methodology was reproducible between examiners and, when combined with good controls and multilevel statistical modelling, was able to discriminate a single application of desensitizing agent.

Humans↗

Single-step adhesives are permeable membranes.

OBJECTIVES: This study tested the hypotheses that micro-tensile bond strengths of all currently available single-step adhesives to dentine are adversely affected by delayed activation of a light-cured composite, and that such a phenomenon only occurs in the presence of water from the substrate side of the bonded interface. METHODS: In experiment I, a control three-step adhesive (All-Bond 2, Bisco) and six single-step adhesives (One-Up Bond F, Tokuyama; Etch&Prime 3.0, Degussa; Xeno CF Bond, Sankin; AQ Bond, Sun Medical; Reactmer Bond, Shofu and Prompt L-Pop, 3M ESPE) were bonded to sound, hydrated dentine. A microfilled composite was placed over the cured adhesive and was either light-activated immediately, or after leaving the composite in the dark for 20 min. In experiment II, three single-step adhesives (Etch&Prime 3.0, Xeno CF Bond and AQ Bond) were similarly bonded to completely dehydrated dentine using the same delayed light-activation protocol. In experiment III, a piece of processed composite was used as the bonding substrate for the same three single-step adhesives. The microfilled composite was applied to the cured adhesives using the same immediate and delayed light-activation protocols. Bonded specimens were sectioned for micro-tensile bond strength evaluation. Fractographic analysis of the specimens was performed using SEM. Stained, undemineralised sections of unstressed, bonded specimens were also examined by TEM. RESULTS: When bonded to hydrated dentine, delayed light-activation had no effect on the control three-step adhesive, but significantly lowered the bond strengths of all the single-step adhesives (p < 0.05). This adverse effect of delayed light-activation was not observed in the three single-step adhesives that were bonded to either dehydrated dentine or processed composite. Morphological manifestations of delayed light-activation of composite in the hydrated dentine bonding substrate were exclusively located along the composite-adhesive interface, and were present as large voids, resin globules and honeycomb structures that formed partitions around a myriad of small blisters along the fractured interfaces. CONCLUSION: These features resembled the 'overwet phenomenon' that was previously reported along the dentine-adhesive interfaces of some acetone-based three-step adhesives. The cured adhesive layer in single-step adhesives may act as semi-permeable membranes that allow water diffusion from the bonded hydrated dentine to the intermixed zone between the adhesive and the uncured composite. Osmotic blistering of water droplets along the surface of the cured adhesive layer and emulsion polymerisation of immiscible resin components probably account for the compromised bond strength in single-step adhesives after delayed activation of light-cured composites.

Biocompatible Materials↗

Diffusion of resin monomers through human carious dentin in vitro.

The diffusion of 2-hydroxyethylmethacrylate (HEMA) and triethylene glycol dimethacrylate (TEGDMA) from light cured bonding resin-composite resin restorations through human carious dentin was investigated. Extracted human molar teeth with different degrees of caries were obtained from consenting donors. Teeth were classified into three groups according to caries severity (mild, moderate and severe) using subjective criteria. The outer carious lesions were then removed guided by a proprietary caries detector dye. Teeth with exposure of the pulp space after caries removal were excluded from the study. A polypropylene chamber was attached to the cemento-enamel junction of each tooth to contain 1 ml distilled water. Each cavity was restored with a HEMA containing bonding resin then a TEGDMA-containing resin composite. Water samples were retrieved over a time course and analyzed by high performance liquid chromatography. There was great variation between teeth in HEMA and TEGDMA permeability. The cumulative amounts released were of similar magnitude to those observed in non-carious teeth for the mild and moderately-severe groups. However, the cumulative amounts released were markedly greater in severely carious teeth than in those with moderate or mild caries.

Adult↗

The effects of common errors on sealing ability of total-etch adhesives.

OBJECTIVES: This study evaluated the effect of errors commonly made in using total-etch adhesives, on the resulting bond strength, fluid movement and nanoleakage of resin dentin bonds. METHODS: Two total-etch adhesives were used for bonding to dentin according to the manufacturers' recommendations, with meticulous solvent evaporation (control), or with the introduction of common bonding errors-wet bonding without solvent evaporation (no evaporation), and dry bonding. RESULTS: The 24-hour bond strength of the control was significantly higher than the other groups (p <0.05). For all groups, the higher initial permeability declined significantly after 24 h. The fluid movement across bonded dentin was similar in the control and dry bonding for both adhesives, whereas significantly higher permeability (p<0.05) was recorded for the no evaporation groups even after 24 h. Extensive silver impregnation within hybrid layers was seen by TEM in the no evaporation and dry bonding specimens after 24 h. Dry bonding caused collapse of the collagen matrix and interfered with resin infiltration. In contrast, inadequate solvent evaporation and/or residual water during dentin bonding results in dilution or incomplete polymerization of the resin, leading to severe nanoleakage formation. SIGNIFICANCE: Increased permeability associated with incomplete solvent evaporation in total-etch adhesives may lead to poor bond strength.

Acid Etching, Dental↗

Factors contributing to the incompatibility between simplified-step adhesives and self-cured or dual-cured composites. Part II. Single-bottle, total-etch adhesive.

PURPOSE: As adhesives containing hydrophilic/acidic resin components are vulnerable to water movement after polymerization, this study tested the hypothesis that coupling of a single-bottle adhesive (OptiBond Solo Plus) to self/dual-cured composites is compromised by adhesive permeability, even with the adjunctive use of chemical co-initiators. MATERIALS AND METHODS: Two versions of chemical co-initiators (activators) were investigated: the proprietary resin-containing OptiBond Solo Plus Activator (A), and a resin-free solution of 2% benzene sulphinic acid sodium salt in ethanol (B). For microtensile bond testing, hydrated (H) or dehydrated (D) bonded human dentin were coupled to a dual-cured composite (Bis-Core) under light- (L) or self-activation (C) mode. A delayed light-activation mode (DL) was also employed to simulate the slower rate of polymerization of self-cured composites but without the influence from adverse chemical interaction. Nine groups were tested: 1) L-H (control); 2) DL-H; 3) DL-D; 4) C-H; 5) C-D; 6) CA-H; 7) CA-D; 8) CB-H; and 9) CB-D. For transmission electron microscopy, a light-cured and an experimental self-cured composite of the same composition were used for the nine groups. RESULTS: Only the bond strength results of the experimental groups DL-D (Group 3) and CB-D (Group 9) were not significantly different from the control group L-H (p > 0.05). TEM revealed the presence of discrete silver-filled water blisters along the adhesive-composite interface in groups 2, 6, and 8, and within the composite in group 4. Adverse chemical interaction in groups 4 and 5 resulted in the observation of a line of silver deposits along the adhesive composite interface. CONCLUSION: The coupling of composites after prolonged contact with hydrated dentin bonded with OptiBond Solo Plus is affected by the intrinsic permeability of the adhesive. The adjunctive use of the Activator is only slightly effective in improving the coupling of this adhesive with self/dual-cured composites. Although the use of resin-free benzene sulphinic acid sodium salt solution completely eliminates the adverse chemical interaction, the inherent permeability of the polymerized adhesive precludes optimal coupling of self/dual-cured composites to bonded hydrated dentin.

Acid Etching, Dental↗

[The penetration of various adhesives into early enamel lesions in vitro].

The aim of the present study was to evaluate the penetration depth (PD) and the thickness of the oxygen inhibition layer (OIL) of a fissure sealant (Helioseal, Vivadent) and various adhesives (Heliobond, Excite, Vivadent; Resulcin, Merz; Solobond M, Voco; Prompt L-Pop, 3M-Espe) applied to enamel lesions in vitro. From 27 bovine teeth 54 enamel specimens were prepared and covered with nail varnish (control) thus obtaining three windows for treatment. After demineralisation (pH 5.0, 14 d) two of the windows were etched with phosphoric acid (20%, 5 s), whereas the third area served as control. The specimens were divided randomly into six groups (n = 9) and the respective adhesive was applied (90 s), either once or twice. Light-curing followed each application. Enamel slabs (perpendicular to the surface) were cut and studied after infiltration with a fluorescent low-viscous resin using confocal microscopy (CLSM). The image of the lesion was divided into two areas with different grey values. Lesion depths were calculated (ImageJ) from the surface to that point in the lesion where the grey value clearly changed to a darker grey value. The zone with the darker grey values marked the front of demineralisation. Mean lesion depths (+/- SD) after demineralisation were measured at 105 (+/- 21) microm. After single application, Resulcin [89 (+/- 22)%] and Helioseal [98 (+/- 6)%] had almost completely penetrated the lesion. Heliobond [126 (+/- 33)%] and Excite [184 (+/- 40)%] penetrated even deeper than the defined lesion. For Excite double application decreased the OIL significantly (p = 0.03; adjusted paired t-test). Adhesives are capable to penetrate artificial initial enamel lesions completely. Follow-up studies are needed to confirm this effect for natural lesions.

Acrylates↗

In vitro study of the effects of Nd:YAG laser irradiation on the apical sealing of endodontic fillings performed with and without dentin plugs.

OBJECTIVE: The aim of this study was to evaluate the effects of Nd:YAG laser irradiation on the apical sealing of root canal fillings with or without a dentin plug. BACKGROUND DATA: Previous studies have shown that Nd:YAG laser irradiation of root canal wall dentin promotes its melting, recrystallization, and therefore a less permeable surface. MATERIALS AND METHODS: Thirty extracted and endodontically treated teeth were divided into three groups. Eight specimens of each group were evaluated by means of quantitative analysis, measuring the marginal apical dye leakage. The remaining two specimens were evaluated by means of qualitative analysis, using the scanning electron microscopy (SEM). For the quantitative analysis, the samples of group I were obturated. In group II, the apical third of the samples were irradiated by the Nd:YAG laser at four exposures of 100 mJ/pulse, 1 W, 10 Hz, 3 sec, and obturated. In group III, an apical dentin plug was condensed and lased using the same parameters of group II, and the root canals were obturated. The samples evaluated by SEM were prepared as those analyzed quantitatively; however, their root canals were not filled. RESULTS: There were statistically significant differences among the groups regarding dye leakage. Group II showed the minor leakage average, followed, in order, by groups I and III. SEM analysis revealed open dentinal tubules in group I; melting and recrystallizing of the apical root dentin in group II; and melting and recrystallizing of a portion of the dentin plug in group III. CONCLUSION: The Nd:YAG laser irradiation of the root canal apical third without the presence of dentin plug significantly improved the quality of the apical sealing under the experimental conditions tested.

Dental Pulp Cavity↗

Single-step, self-etch adhesives behave as permeable membranes after polymerization. Part II. Silver tracer penetration evidence.

PURPOSE: To investigate the existence of potential water-filled channels in resin-dentin interfaces of some single-step, self-etch adhesives. METHODS: In Experiment I, Prompt L-Pop (3M ESPE) or an experimental adhesive RZ II (Sun Medical) were bonded to hydrated dentin and then coupled with a light-cured composite that was light-activated immediately upon application. In Experiment II, RZ II and FuturaBond (Voco) were similarly bonded to hydrated dentin. Unfilled resins were coupled in the dark to the bonded dentin using the chemical-cured mode, or light-cured mode with a 20-minute delay before light-activation. Bonded assemblies were challenged with ammoniacal silver nitrate as a tracer and examined with transmission electron microscopy. RESULTS: In Experiment I, two types of silver patterns were observed within adhesive layers and hybrid layers: interconnecting reticular, finger-like and tree-like patterns that were collectively named "water trees", and discrete silver grains. In Experiment II, apart from the patterns described above, silver-filled water blisters were also observed between the adhesive and the unfilled coupling resin in RZ II. They were linked to the pre-existing "water trees" in the adhesive layers via multiple, short water bridges. In FuturaBond, similar blisters were located between the first adhesive layer and the second adhesive layer that was light-activated after placement of the coupling resin.

Acid Etching, Dental↗

Effect of new generation surface sealants on the marginal permeability of Class V resin composite restorations.

This study evaluated, in vitro, the effectiveness of 5 surface sealants (Biscover, Optiguard, Seal-n-Shine, PermaSeal and DuraFinish); 1 pit and fissure sealant (Helioseal) and 1 dentin bonding agent (Adper Scotchbond MultiPurpose Adhesive) on the marginal sealing ability of Class V hybrid resin composite (Esthet.X) restorations. Ninety-six non-carious human molars were randomly assigned to 8 groups (n=12). Class V cavities were prepared on either the facial or lingual surfaces, with coronal margins in enamel and apical margins in cementum (dentin), and restored following manufacturers' instructions. Following finishing and polishing procedures, the covering agents were applied to each restoration and adjacent tooth surface, except for the control group restorations, which were not sealed. The teeth were thermocycled, immersed in a 1% methylene blue dye solution, sectioned and analyzed for dye penetration (leakage) using a 20x binocular microscope. Microleakage was evaluated at the coronal and apical margins using an 0-3 ordinal grading scale. Non-parametric data was analyzed at a p< or =0.05 level of significance. The Kruskal-Wallis test showed a significant difference was exhibited among groups at the coronal margins, with Helioseal pit/fissure sealant and DuraFinish surface sealant exhibiting significantly less leakage than the control and Adper Scotchbond MultiPurpose adhesive. At the apical margins, DuraFinish surface sealant showed significantly less leakage than the Biscover, Seal-n-Shine and PermaSeal surface sealants or Helioseal pit/fissure sealant, Adper Scotchbond MultiPurpose adhesive and the control group. According to the Wicoxon signed-rank test, significantly greater leakage was revealed at the apical margins compared to the enamel margins of the material groups.

Acrylates↗

Effect of dentin primer on shear bond strength of composite resin to moist and dry enamel.

The etched enamel-composite resin bond is the most reliable bond known to us. Moisture and dentin primers are the two most important variables that can interfere with this bond. This study investigated the effect of dentin primer on bond strengths of composite resin to moist and dry enamel. One hundred freshly extracted molar teeth were used for shear bond strength testing. The teeth were mounted in phenolic rings with an approximal enamel surface exposed. The exposed enamel surface on each tooth was flattened using 320- 400- and 600-grit silicon carbide papers and etched using 34-38% phosphoric acid gel. The teeth were then divided into 10 groups (n = 10). Four groups were assigned to each of the two dentin bonding systems, Scotchbond Multi-Purpose and OptiBond FL. Two groups were assigned to the single-bottle bonding agent (Single Bond). Each bonding system was tested on moist and dry enamel. OptiBond FL and Scotchbond MP were tested with and without the use of primer. All samples were thermocycled and tested in shear. Fracture analysis was performed using a binocular microscope. For scanning electron microscopy, approximal samples of enamel (1 mm thick) were flattened, etched, and bonded with and without primer on moist and dry enamel. A 1 mm-thick layer of Z100 was bonded to the specimens, which were then immersed in 10% HCl for 24 hours to dissolve the enamel. The specimens were viewed under a scanning electron microscope. Results indicated that the use of primer on dry enamel did not significantly affect (P > 0.05) shear bond strengths for the two bonding systems, Scotchbond MP (primed 24.10 +/- 4.83 MPa, unprimed 29.57 +/- 7.49 MPa) and OptiBond FL (primed 26.82 +/- 4.44, unprimed 25.66 +/- 2.95). However, the use of primer was found to be essential on moist enamel to obtain acceptable bond strengths with both Scotchbond MP (primed 25.61 +/- 10.29 MPa, unprimed 3.26 +/- 0.95 MPa) and OptiBond FL (primed 30.28 +/- 3.49 MPa, unprimed 8.37 +/- 3.31 MPa). Moisture on enamel did not significantly affect (P > 0.05) bond strengths for the single-bottle bonding agent, Single Bond (moist enamel 31.34 +/- 9.03 MPa, dry enamel 27.93 +/- 5.41 MPa). Fracture analysis revealed that most fractures were adhesive or mixed, with a greater percentage being cohesive for the groups with dry enamel or with primer on moist enamel. Scanning electron micrographs corroborated the shear bond strength data. The specimens without primer on moist enamel showed very poor penetration of adhesive and composite resin into the etched enamel microporosities.

Analysis of Variance↗

Bonding characteristics of self-etching adhesives to intact versus prepared enamel.

PURPOSE: This study tested the null hypothesis that the preparation of the enamel surface would not affect the enamel microtensile bond strengths of self-etching adhesive materials. MATERIALS AND METHODS: Ten bovine incisors were trimmed with a diamond saw to obtain a squared enamel surface with an area of 8 x 8 mm. The specimens were randomly assigned to five adhesives: (1) ABF (Kuraray), an experimental two-bottle self-etching adhesive; (2) Clearfil SE Bond (Kuraray), a two-bottle self-etching adhesive; (3) One-Up Bond F (Tokuyama), an all-in-one adhesive; (4) Prompt L-Pop (3M ESPE), an all-in-one adhesive; and (5) Single Bond (3M ESPE), a two-bottle total-etch adhesive used as positive control. For each specimen, one half was roughened with a diamond bur for 5 seconds under water spray, whereas the other half was left unprepared. The adhesives were applied as per manufacturers' directions. A universal hybrid composite resin (Filtek Z250, 3M ESPE) was inserted in three layers of 1.5 mm each and light-cured. Specimens were sectioned in X and Y directions to obtain bonded sticks with a cross-sectional area of 0.8 +/- 0.2 mm2. Sticks were tested in tension in an Instron at a cross-speed of 1 mm per minute. Statistical analysis was carried out with two-way analysis of variance and Duncan's test at p < .05. Ten extra specimens were processed for observation under a field-emission scanning electron microscope. RESULTS: Single Bond, the total-etch adhesive, resulted in statistically higher microtensile bond strength than any of the other adhesives regardless of the enamel preparation (unprepared = 31.5 MPa; prepared = 34.9 MPa, not statistically different at p < .05). All the self-etching adhesives resulted in higher microtensile bond strength when enamel was roughened than when enamel was left unprepared. However, for ABF and for Clearfil SE Bond this difference was not statistically significant at p > .05. When applied to ground enamel, mean bond strengths of Prompt L-Pop were not statistically different from those of Clearfil SE Bond and ABF. One-Up Bond F did not bond to unprepared enamel. Commercial self-etching adhesives performed better on prepared enamel than on unprepared enamel. The field-emission scanning electron microscope revealed a deep interprismatic etching pattern for the total-etch adhesive, whereas the self-etching systems resulted in an etching pattern ranging from absent to moderate.

Acid Etching, Dental↗