In vitro effect of nanoleakage expression on resin-dentin bond strengths analyzed by microtensile bond test, SEM/EDX and TEM

This study evaluated the effect of multiple consecutive adhesive resin coatings of adhesive bonded to human dentin on nanoleakage and resin-dentin bond strength. Resin bonded dentin specimens were prepared using a total-etch adhesive (One-Step Plus) applied as multiple consecutive coating, or using two self-etch adhesive systems (iBond or Fluoro Bond). For the total-etch adhesive, resin application and air evaporation were performed 1, 2, 3, or 4 times. The self-etch adhesives were applied according to manufacturers' instructions. Resin-dentin bonded beams were prepared and immersed in water (control) or ammoniacal silver nitrate. After storage, microtensile bond strengths were measured. The fractured surfaces were examined by scanning and transmission electron microscopy (SEM and TEM), and energy-dispersive X-ray spectrometry (EDX). No significant differences in bond strength were found between water and silver nitrate storage groups. Several types of silver depositions (spotted, reticular, or water trees) were found in adhesive joints. The bond strengths of the single coated specimens of the total-etch adhesive were significantly lower than those receiving 2-4 coatings. Single coats produced more nanoleakage than multiple coats. However, no correlation was found between the bond strengths and nanoleakage between the different adhesives (total-etch adhesive with different conditions or self-etch adhesives).     

Micromorphological changes in resin-dentin bonds after 1 year of water storage

The purpose of this study was to evaluate the degradation of resin-dentin bonds after 1 year of water storage. Resin-dentin-bonded specimens were prepared with the use of an adhesive resin system (One-Step: Bisco). Half of the experimental specimens were sectioned perpendicular to the adhesive interface to produce a beam (adhesive area: 0.9 mm(2)) before being stored in distilled water at 37 degrees C for 1 year. The remaining half of the bonded specimens were sectioned into beams of similar dimensions after 1 year of water storage. Additional bonded specimens that had been stored in water for 24 h before sectioning into beams were used as controls. The beams in the two experimental groups and the control group were subjected to microtensile bond testing. Fractography was performed on all fractured beams with the use of FE-SEM. There were significant (p <.05) differences in bond strength among the control specimens (55.9 +/- 12.9 MPa), specimens that had been sectioned into beams after water storage (68.9 +/- 18.6 MPa), and specimens that had been sectioned into beams before water storage (28.1 +/- 9.3 MPa). Fractography revealed that the resin material was gradually extracted from the periphery to the center portion of the beam. This probably accounted for the decrease in bond strength after 1 year of water storage.     

Effects of a ferric chloride primer on collagen-depleted dentin bonding between tri-n-butylborane initiated self-curing resin and dentin

This study was designed to evaluate the micro-tensile bond strength between a carboxylic resin and dentin, when the dentin surface was modified with an experimental dentin primer. The three primers tested were ED primer II (ED), 0.3% ferric chloride aqueous solution (FE), and ED containing 0.3% ferric chloride (ED/FE). Three commercial dentin conditioners [40% phosphoric acid, 10% NaOCl, and 10% citric acid with 3% ferric chloride (10-3)] were also used. The coronal surfaces of extracted human molars were ground flat to dentin. The dentin surfaces were treated with phosphoric acid, NaOCl, or with one of the primers. The 10-3 was used without phosphoric acid or NaOCl as a control. A composite material rod was bonded to the dentin surface with 4-META/MMA-TBB resin. After 24-h immersion in 37 degrees C distilled water, 0.9 mm x 0.9 mm composite-dentin beams cut from the bonded specimens were stressed to failure in tension at 0.6 mm/min. The bond strengths were also evaluated after 5000 thermocycles. The bond strength of the group ED/FE was significantly higher than those of the 10-3, ED, and FE. After 5000 thermocycling, 10-3, ED and FE showed significant decrease in bond strength, although no significant decrease was seen for ED/FE. It was concluded that dentin surface treatment with phosphoric acid, NaOCl, and the ED/FE primer improved the bonding (p < 0.05) between 4-META/MMA-TBB resin and dentin, with or without thermocycling, while the bond strengths in the control group fell 34% following 5000 thermocycles.     

In vitro degradation of resin-dentin bonds analyzed by microtensile bond test,scanning and transmission electron microscopy

Our knowledge of the mechanisms responsible for the degradation of resin-dentin bonds are poorly understood. This study investigated the degradation of resin-dentin bonds after 1 year immersion in water. Resin-dentin beams (adhesive area: 0.9mm(2)) were made by bonding using a resin adhesive, to extracted human teeth. The experimental beams were stored in water for 1 year. Beams that had been stored in water for 24h were used as controls. After water storage, the beams were subjected to microtensile bond testing. The dentin side of the fractured surface was observed using FE-SEM. Subsequently, these fractured beams were embedded in epoxy resin and examined by TEM. The bond strength of the control specimens (40.3+/-15.1MPa) decreased significantly (p<0.01) after 1 year of water exposure (13.3+/-5.6MPa). Loss of resin was observed within fractured hybrid layers in the 1 year specimens but not in the controls. Transmission electron microscopic examination revealed the presence of micromorphological alterations in the collagen fibrils after 1 year of water storage. These micromorphological changes (resin elution and alteration of the collagen fibrils) seem to be responsible for the bond degradation leading to bond strength reduction.     

Differential effect of in vitro degradation on resin-dentin bonds produced by self-etch versus total-etch adhesives

OBJECTIVE: To evaluate the effect of an in vitro challenge (NaOCl immersion) on microtensile bond strength (MTBS) of five adhesive systems to dentin. METHODS: Flat dentin surfaces from 40 molars were bonded with three total-etch adhesives (Single Bond, Prime&Bond NT and the experimental Prime&Bond XP), and two self-etching agents (Clearfil SE Bond and Etch&Prime 3.0). Composite build-ups were constructed with Tetric Ceram. Teeth were then sectioned into beams of 1.0 mm2 cross-sectional area. Half of the beams were immersed in 10% NaOCl aqueous solution for 5 h. Each beam was tested in tension in an Instron machine at 0.5 mm/min. Data were analyzed by 2-way ANOVA and multiple comparisons tests (p < 0.05). RESULTS: Clearfil SE Bond and Single Bond attained higher MTBS than the other three adhesives. Prime&Bond NT and Prime&Bond XP performed equally, and Etch&Prime resulted in the lowest MTBS. After NaOCl immersion, MTBS decreased in all groups. The highest MTBS values were obtained for Clearfil SE Bond and Prime&Bond XP. Scaning electron microscopy observation of debonded sticks evidenced dissolution and microstructural alterations of intertubular dentin, except when Clearfil SE Bond was used. CONCLUSIONS: Resin-dentin bonds are prone to chemical degradation. The extent of the resin degradation is adhesive system specific. Chemical degradation of the nonresin infiltrated collagen fibers does also exist in total-etch adhesives. Both processes may reduce long-term resin-dentin bond strength.     

Effect of load cycling and in vitro degradation on resin-dentin bonds using a self-etching primer

The objective of this study was to evaluate the effect of in vitro degradation and mechanical loading on microtensile bond strength (MTBS) and microleakage (ML) of a resin composite to dentin using a self-etching primer adhesive [Clearfil SE Bond (SEB)] under two hydration statuses. Twenty-four flat dentin surfaces were divided in groups: 1) blot-dried, 2) air-dried. SEB was applied and resin buildups were performed with Tetric Ceram. Specimens were divided in four subgroups: a) sectioned into beams, b) load cycled, c) beams were immersed in NaOCl for 5 h, d) load cycled and immersed in NaOCl. Beams were tested in tension. For ML testing, 80 Class V cavities were prepared and molars divided in subgroups as described above (in group c and d, specimens were kept in distilled water for 1 year, instead of the 5-h NaOCl immersion). ML was assessed by dye penetration. Analysis of variance and multiple comparisons tests were used for MTBS. For ML, Mann-Whitney U and Wilcoxon matched pairs signed ranked were used (p<0.05). SEB applied to completely dehydrated dentin produced the highest MTBS, at 24-h evaluation. In vitro degradation always decreased MTBS, and fatigue loading only diminished MTBS on dehydrated dentin. Load cycling increased dye penetration on dentin margins. Degradation always increased ML in both enamel and dentin margins.     

Micro-rotary fatigue of tooth-biomaterial interfaces

The bonding effectiveness of restorative materials to tooth tissue is typically measured statically. Clinically tooth/composite bonds are however subjected to cyclic sub-critical loads. Therefore, in vitro fatigue testing of dental adhesives should predict better the in vivo performance of adhesives. The objective of this study was to determine the fatigue resistance of two representative adhesives, a self-etch and an etch&rinse adhesive, bonded to enamel and dentin. Therefore, tooth/composite interfaces were cyclically loaded using a miniaturized version of a rotating beam fatigue testing device. Subsequently, the load at which 50% of the specimens fail after 10(5) cycles, was determined as the median micro-rotary fatigue resistance (microRFR). For both adhesives, the microRFR was about 30-40% lower than the corresponding micro-tensile bond strength (microTBS) to both enamel and dentin. Analysis of the fracture surfaces by Feg-SEM revealed typical fatigue fracture patterns. It is concluded that resin/tooth interfaces are vulnerable to progressive damage by sub-critical loads, with the 3-step etch&rinse adhesive being more resistant to fatigue than the 2-step self-etch adhesive.     

Nanoleakage and microshear bond strength in deproteinized human dentin

This study evaluated the influence of dentin deproteinization with NaOCl on the microshear bond strength (microSBS) and the nanoleakage patterns of three dentin bonding systems (DBS). Occlusal dentin surfaces, obtained from extracted noncarious human molars, were divided into two experimental groups, according to dentin surface treatment: Group I-37% H(3)PO(4)/15s and Group II-37% H(3)PO(4)/15s + 10% NaOCl/1 min. The dentin surfaces were bonded with one of the following DBS: Scotchbond Multipurpose-SBMP, Prime & Bond NT-PB and Clearfil SE Bond-SE. After 1 week storage in water at 37 degrees C, the specimens were subjected to the microSBS test. The data were analyzed by two-way ANOVA and Student-Newman-Keuls' test (p = 0.05). The nanoleakage was evaluated using scanning electron microscopy (SEM) in backscattered electron imaging regime. No significant difference in microSBS between dentin treatments was found for SBMP. For PB, microSBS increased after NaOCl dentin treatment. SE showed a reduction in microSBS in deproteinized specimens. SEM analysis showed different nanoleakage patterns for each DBS. Irrespective of dentin treatments, all SBMP specimens showed nanoleakage. SE did not show nanoleakage with the two dentin treatments. PB showed nanoleakage within the hybrid layer only in acid-etched specimens. The influence of dentin deproteinization was dependent on the dentin bonding system formulation.     

In vitro microTBS of one-bottle adhesive systems: sound versus artificially-created caries-affected dentin

This in vitro study aimed to evaluate a pH-cycling model for simulation of caries-affected dentin (CAD) surfaces, by comparing the bond strength of etch-and-rinse adhesive systems on sound and artificially-created CAD. Dentin substrates with different mineral contents and morphological patterns were created by submitting buccal bovine dentin to the following treatments: (1) immersion in artificial saliva during the experimental period (sound dentin, SD), or (2) induction to a CAD condition by means of a dynamic pH-cycling model (8 cycles, demineralization for 3 h followed by mineralization for 45 h). The bond strength of Excite or Prime and Bond NT adhesive systems was assessed using the microtensile bond strength (microTBS) test. Dentin microhardness was determined by cross-sectional Knoop evaluations. Resin-dentin morphology after the treatments was examined by scanning electron microscopy. SD produced significantly higher microTBS than CAD for both adhesives evaluated, without differences between materials. CAD exhibited lower microhardness than SD. Morphological analysis showed marked distinctions between SD and CAD bonded interfaces. Under the conditions of this study, differences in morphological pattern and dentin mineral content may help to explain resin-dentin bond strengths. The proposed pH-cycling model may be a suitable method to simulate CAD surfaces for bonding evaluations.     

Microtensile bond strength of etch and rinse versus self-etch adhesive systems

The aim of this study was to compare the microtensile bond strength of the etch and rinse adhesive versus one-component or two-component self-etch adhesives. Twelve intact human molar teeth were cleaned and the occlusal enamel of the teeth was removed. The exposed dentin surfaces were polished and rinsed, and the adhesives were applied. A microhybride composite resin was applied to form specimens of 4?mm height and 6?mm diameter. The specimens were sectioned perpendicular to the adhesive interface to produce dentin-resin composite sticks, with an adhesive area of approximately 1.4?mm(2). The sticks were subjected to tensile loading until failure occurred. The debonded areas were examined with a scanning electron microscope to determine the site of failure. The results showed that the microtensile bond strength of the etch and rinse adhesive was higher than that of one-component or two-component self-etch adhesives. The scanning electron microscope examination of the dentin surfaces revealed adhesive and mixed modes of failure. The adhesive mode of failure occurred at the adhesive/dentin interface, while the mixed mode of failure occurred partially in the composite and partially at the adhesive/dentin interface. It was concluded that the etch and rinse adhesive had higher microtensile bond strength when compared to that of the self-etch adhesives.     

Bonding characteristics of newly developed all-in-one adhesives

This study evaluated the microtensile bond strength and the interfacial morphology of newer adhesives. The occlusal surfaces of extracted teeth were ground flat for random allocation to four equal groups. Resin composite was bonded to each surface using either Clearfil SE Bond [SEB], Clearfil Protect Bond [PB], G-Bond [GB], or an experimental adhesive, SSB-200 [SSB]. After storage for 24 h in water at 37 degrees C, they were sectioned into beams (cross-sectional area 1 mm(2)) for microtensile bond strength testing (muTBS) at a crosshead speed of 1 mm/min. The load at failure of each was recorded; the data were analyzed by one-way ANOVA and Games Howell tests. The surfaces of the fractured specimens were observed using SEM. For the ultra-morphology of the interface, the occlusal surfaces of four more teeth were prepared as before and a thin layer of flowable resin composite was bonded to each surface using one of the four adhesives.The mean muTBS ranged from 39.68 MPa (GB) to 64.97 MPa (SEB). There were no statistical differences between SEB and SSB, or between PB and GB (p > 0.05). The muTBS of SEB and SSB were significantly greater than that of PB and GB (p < 0.05). SEMs of the fractured surfaces revealed a mixed (cohesive/interfacial) failure. TEM examination highlighted differences in the hybrid layer; SEB had a thicker layer than the others. In conclusion, the newer all-in-one adhesives produced a thin hybrid layer but varied in their bond strengths. The 2-step self-etching adhesives do not necessarily produce higher bond strengths than that of the all-in-one systems.     

The effect of two configuration factors,time, and thermal cycling on resin to dentin bond strengths

Most in vitro testing of bonding systems is performed using specimens made in a mold with a low configuration (C) factor (ratio of bonded/unbonded surfaces) whereas clinically the C-factor is usually much greater. This study compared the effect of thermal cycling on the measured shear bond strength of 3M Single Bond dental adhesive bonded to dentin using molds with two different C-factors. The hypothesis was that neither C-factor nor thermal cycling would affect measured bond strengths. Resin composite was bonded to human dentin in cylindrical molds with an internal diameter of 3.2mm and either 1mm or 2.5mm deep. The 1mm deep molds had a C-factor of 2.2 and the 2.5mm deep molds had a C-factor of 4.1. Specimens were debonded either 10min after they had been bonded to dentin, or after they had been stored for 7 days in water at 37+/-1 degrees C, or after thermal cycling 5000 times for 7 days. Two-way ANOVA showed that overall both the C-factor and the storage condition had a significant effect on bond strength (p<0.001). There was a significant interaction (p<0.001) between the C-factor and how the specimens had been stored. The GLM/LSMEANS procedure with Sidak's adjustment for multiple comparisons showed that overall the specimens made in the mold with a high C-factor (4.1) had a lower bond strength than those that had been made in the mold with a lower (2.2) C-factor (p<0.001). Thermal cycling had a negative effect on the bond strength only for specimens made in molds with a C-factor of 4.1 (p<0.001).     

Hardness, elasticity, and ultrastructure of bonded sound and caries-affected primary tooth dentin

Biomechanical properties of bonded dentin are important factors for resin restoration. We evaluated the hardness and elastic modulus of bonded sound and caries-affected primary tooth dentin using a one-step adhesive system, and observed the microstructure of the bonded interface. Six sound and six carious primary teeth were used. For sound teeth, flat occlusal dentin surfaces were prepared with a water-cooled high-speed diamond bur. For carious teeth, infected dentin was stained with a caries detector and removed with a water-cooled low-speed round steel bur and hand instruments. The prepared dentin was bonded with One-Up Bond F Plus (Tokuyama Dental Co., Tokyo, Japan). The resin-dentin interface and dentin beneath the interface were measured with a nano-indentation tester and observed with SEM and TEM. For both the carious and sound teeth, there was no significant difference between the hardness of the interfacial dentin and dentin 10-80 microm beneath the interface. However, the Young's modulus of the interfacial dentin was significantly lower than the dentin 40-80 microm (carious teeth) or 50-80 microm (sound teeth) beneath the interface. Both the hardness and Young's modulus of the interfacial dentin were not significantly different between the carious and sound teeth. Compared to the sound dentin, the hybrid layer on the caries-affected dentin was thicker and exhibited more complicated morphologic features. The thickness of the hybrid layers was generally less than 1 microm.     

Effect of sodium hypochlorite on dentin bonding with a polyalkenoic acid-containing adhesive system

The purpose of this study was to evaluate the effect of sodium hypochlorite (NaOCl) treatment on dentin bonding by means of contact angle (CA), shear bond strength (SBS), and microleakage (ML) measurements. Ultrastructure and nanoleakage (NL) of the interfaces were examined by transmission electron microscopy (TEM). For CA, SBS, and TEM evaluation, human molars were sectioned to expose dentin surfaces and were either acid-etched (35% H3PO4) or further treated with 5% NaOCl for 2 min before the application of Single Bond adhesive. CAs were measured using the Axisymmetric Drop Shape Analysis technique. The Watanabe testing assembly was used for SBS evaluation. ML was assessed by a dye penetration method. NL was examined using a silver-staining technique. The results showed that CA values decreased after acid etching and even more after NaOCl treatment. NaOCl treatment produced lower SBS than acid-etched dentin. Both ML values and NL manifestations were similar for NaOCl-treated and acid-etched dentin. NaOCl did not completely remove the collagen matrix. NL was manifested along the base of hybrid layers and within the polyalkenoic acid copolymer in both groups. Adverse chemical interactions could have occurred between the remnant collagen matrix and/or mineralized dentin after NaOCl treatment. There is no additional advantage in using NaOCl treatment with this adhesive.     

Evaluation of the fatigue behavior of the resin-dentin bond with the use of different methods

The clinical performance of directly bonded resin composites is fundamentally dependent on durable adhesion to prevent gap formation over time. The goal of this investigation was to evaluate the effectiveness of various dentin adhesives by means of quasistatic and dynamic dentin bond strengths, and also to determine marginal and internal gap formation after loading in an artificial oral environment. Three hundred thirty human third molars were used within four weeks of extraction. Adhesives used were A.R.T. Bond, OptiBond FL, Scotchbond Multi-Purpose Plus, Single Bond, Prime & Bond NT, and One Up Bond F for bonding of one resin composite (Z 250). Buccal and lingual aspects of 90 teeth were ground flat to expose dentin, then resin composite cylinders were bonded. Initial bond strengths (n = 10) and adhesive fatigue limits (n = 20) were determined with the use of a shear test apparatus. One hundred eighty conical cavities were prepared into dentin discs and filled with the same materials. After 21 days of storage, initial push-out bond strengths (n = 10) and adhesive fatigue limits (n = 20) were measured. Sixty molars with MO cavities (n = 10) with margins below the cement-enamel junction were filled. Before and after thermomechanical loading (100000 x 50 N and 2500 x thermocycling between + 5 and + 55 degrees C), marginal gap formation and internal adaptation (only after loading) were analyzed under a SEM (x 200). The one-bottle systems showed higher shear bond strengths when evaluated statically and dynamically. However, cyclic fatigue push-out bond strengths resulted in higher values for older multistep systems. Marginal and internal gap analysis confirmed the results, in favor of older adhesive systems (p <.05; Mann-Whitney U test).     

Adhesion of a self-etching system to dental substrate prepared by Er:YAG laser or air abrasion

The purpose of this study was to assess the microtensile bond strength of a self-etching adhesive system to enamel and dentin prepared by Er:YAG laser irradiation or air abrasion, as well as to evaluate the adhesive interfaces by scanning electron microscopy (SEM). For microtensile bond strength test, 80 third molars were randomly assigned to five groups: Group I, carbide bur, control (CB); II, air abrasion with standard tip (ST); III, air abrasion with supersonic tip (SP); IV, Er:YAG laser 250 mJ/4 Hz (L250); V, Er:YAG laser 300 mJ/4 Hz (L300). Each group was divided into two subgroups (n = 8) (enamel, E and dentin, D). E and D surfaces were treated with the self-etching system Adper Prompt L-Pop and composite buildups were done with Filtek Z-250. Sticks with a cross-sectional area of 0.8 mm(2) (+/-0.2 mm(2)) were obtained and the bond strength tests were performed. Data were submitted to ANOVA and Tukey's test. For morphological analysis, disks of 30 third molars were restored, sectioned and prepared for SEM. Dentin presented the highest values of adhesion, differing from enamel. Laser and air-abrasion preparations were similar to enamel. Dentin air-abrasion with standard tip group showed higher bond strength results than Er:YAG-laser groups, however, air-abrasion and Er:YAG laser groups were similar to control group. SEM micrographs revealed that, for both enamel and dentin, the air-abrasion and laser preparations presented irregular adhesive interfaces, different from the ones prepared by rotary instrument. It was concluded that cavity preparations accomplished by both Er:YAG laser energies and air abrasion tips did not positively influence the adhesion to enamel and dentin.     

Deproteinized dentin: a favorable substrate to self-bonding resin cements?

The adhesive performance on deproteinized dentin of different self-adhesive resin cements was evaluated through microtensile bond strength (μTBS) analysis and scanning electron microscopy (SEM). Occlusal dentin of human molars were distributed into different groups, according to the categories: adhesive cementation with two-step bonding systems-control Groups (Adper Single Bond 2 + RelyX ARC/3M ESPE; One Step Plus + Duolink/Bisco; Excite + Variolink I/Ivoclar Vivadent) and self-adhesive cementation-experimental groups (Rely X Unicem/3M ESPE; Biscem/Bisco; MultiLink Sprint/Ivoclar Vivadent). Each group was subdivided according to the dentin approach to: α, maintenance of collagen fibers and β, deproteinization. The mean values were obtained, and submitted to ANOVA and Tukey test. Statistical differences were obtained to the RelyX Unicem groups (α = 13.59 MPa; β = 30.19 MPa). All the BIS Group specimens failed before the mechanical tests. Dentinal deproteinization provided an improved bond performance for the self-adhesive cement Rely X Unicem, and had no negative effect on the other cementing systems studied.     

Effects of sodium hypochlorite gel and sodium hypochlorite solution on dentin bond strength

The aim of this study was to determine the effect of 10% NaOCl gel and 10% NaOCl solution on dentin bond strengths of four adhesive systems. One hundred eighty bovine incisors were ground to achieve a flat polished surface, then divided into 12 groups: Gluma One Bond [G1-control; G2-NaOCl solution; G3-NaOCl gel]; Prime & Bond 2.1 [G4-control; G5-NaOCl solution; G6-NaOCl gel]; Single Bond [G7-control; G8-NaOCl solution; G9-NaOCl gel]; Prime & Bond NT [G10-control; G11-NaOCl solution; G12-NaOCl gel]. Dentin was etched, rinsed, and blot dried. For the experimental groups, after acid etching, 10% NaOCl solution or 10% NaOCl gel was applied for 60 s, rinsed, and blot dried. Composite resin was inserted and light cured. Shear bond strengths were tested with a crosshead speed of 0.5 mm/min. The mean values MPa (SD) were analyzed with two-way ANOVA and Tukey's tests (alpha < 0.01). Ten percent NaOCl solution significantly increased Gluma One Bond strength. No effect was observed for the other adhesives. The 10% NaOCl gel did not affect bond strengths. Ten percent NaOCl gel was less effective on collagen removal as compared to 10% NaOCl solution. The influence of collagen removal on bond strength is dependent on adhesive system, where both the solvent and the monomer can influence the results.     

Microleakage and interfacial morphology of self-etching adhesives in class V resin composite restorations

The purpose of the study was to evaluate the marginal leakage of three adhesive systems in Class V resin composite restorations. Two adhesive systems containing acidic primers: Clearfil SE Bond (CSEB) and Etch & Prime 3.0 (E&P), were compared with a conventional water-based primer: Scotchbond Multipurpose Plus (SBMP). Class V cavities were made at the cementum-enamel junction of extracted human molars, which were then divided between three groups. One of the adhesive systems was applied to each group following manufacturers' instructions. Composite restorations were placed, light cured for 40 s, and polished. Specimens were then immersed in a solution of 2% basic fuchsin dye for 24 h. Longitudinal sections were obtained and studied with a stereomicroscope for assessment of the microleakage according to the degree of dye penetration (scale of 0-3). Data were analyzed by Kruskal-Wallis one-way ANOVA, Mann-Whitney tests, and the Wilcoxon matched-pairs signed rank test. Two specimens for each group were analyzed by scanning-electron microscopy (SEM). Bonded interfaces of dentin were also examined by transmission-electron microscopy (TEM). On enamel, there were no significant differences between the three groups. On dentin, CSEB showed the lowest dye penetration values among the three adhesive systems. SEM and TEM studies showed hybrid layer and resin tag formations in all groups.     

Removal of noncollagenous components affects dentin bonding

The structural integrity of fibrillar type I collagen is critical for effective dentin bonding. Since most noncollagenous matrix components in dentin are closely associated with collagen, we hypothesized that they may also contribute to dentin bonding. To test this hypothesis, bovine dentin was acid-etched, treated with chondroitinase ABC (C-ABC), endo-beta-galactosidase (Endo-beta), or trypsin. Controls were prepared in the same manner but without the enzymes. All control and experimental specimens were then bonded with One-Step. Bond strength data were analyzed by one-way ANOVA and Fisher's PLSD test (p < 0.05). When dentin was treated with C-ABC or trypsin, bond strengths significantly decreased for the rewetted groups (p < 0.05). The treatment with Endo-beta showed no effects on bond strengths (p > 0.05). When the treated dentin surfaces were observed under SEM, the C-ABC and trypsin treated groups revealed significant loss of collagen fibril architecture. The results indicate that chondroitin sulfate glycosaminoglycans and trypsin-digestible noncollagenous proteins play roles in maintaining the open dimensions of the collagen fibril scaffold, which is essential for optimal dentin bonding.