Effect of solvent content on resin hybridization in wet dentin bonding

With wet bonding techniques, the channels between the demineralized dentin collagen fibrils are filled with debris, solvent, and water. Commercial adhesives include solvents such as ethanol or acetone to facilitate resin-infiltration into this wet substrate. Under in vivo conditions, the solvent may be diluted because of repeated exposure of the material to the atmosphere, or concentrated because of separation of the bonding liquids into layers within the bottle. The purpose of this study was to investigate the effect of different concentrations of ethanol (10-50%) on infiltration of the adhesive resin and collagen fibril encapsulation in the adhesive/dentin interface using light microscopy, micro-Raman spectroscopy, and scanning electron microscopy. The results indicated that under wet bonding conditions the hybridization process was highly sensitive to the initial solvent concentration in the adhesive system. The staining and scanning electron microscopy results showed that the quality of the interfacial hybrid layer was poor at the lower (10%) or higher (50%) ethanol content. Micro-Raman analysis indicated that there was a distinct difference in the degree of adhesive penetration among adhesives containing different concentrations of ethanol. Adhesives containing 10 or 50% ethanol did not realize effective penetration; the penetration of the adhesive monomers increased dramatically when the initial ethanol content was 30%. The amount of solvents are essential for achieving effective bonding to dentin.     

Recent advances in the theory and mechanism of adhesive resin bonding to dentin: a critical review

Dentin bonding issues involving adhesive resins have attracted considerable research interest in recent years. An important advance due to the ongoing research is the concept of hybridization of the tissue with primer/adhesive systems. Hybridization involves permeation of primer monomer into the tissue substrate. Although the mechanism of adhesive permeation and interaction with tissue may be complex, significant advances have been made. In systems where etching precedes priming and bonding steps, the Hoy's solubility parameter compatibility of the primer formulation with that of demineralized dentin matrix may determine adhesive permeability. Monomer permeation brings the primer atoms in closer contact with the substrate atoms, leading to adhesive interactions through van der Waals, hydrogen bonding, and electrostatic interactions. In self-etch primer systems, stronger electrostatic interaction between primer monomers and hydroxyapatite has been used to explain the adhesion process. These interactions have been computer-modeled and analyzed. Such interactions and subsequent polymerization of the monomer promote improved bond strength and efficient margin sealing. Incomplete permeation of monomer into the full depth of demineralized region may, however, leave exposed collagen fibrils and cause nanoleakage of water into these regions through a 20-100 nm sized marginal gap, leading to subsequent hydrolytic degradation of these collagen fibrils and the hybrid layer. Microleakage is also a problem in some single step formulations. In this review, we analyze these current theoretical and mechanism-related issues of interest in adhesive resin bonding to dentin, and outline the continuing problems that need to be overcome in the future.     

Infiltration/evaporation-induced shrinkage of demineralized dentin by solvated model adhesives

During dentin bonding, solvated adhesive comonomers are applied to water-saturated decalcified dentin matrices. When alcohol-solvated hydrophilic or hydrophobic methacrylate monomers are applied, they chemically remove water and cause matrix shrinkage during comonomer infiltration. Evaporation of solvent induces further shrinkage. The purpose of this work was to compare the shrinkage of water-saturated dentin matrices infiltrated with ethanol- or methanol-solvated 2-hydroxyethyl methacrylate (HEMA), 2,2-bis[4(2-hydroxy-3-methacryloyloxy-propyloxy)-phenyl] propane (BisGMA), or triethyleneglycol dimethacrylate (TEGDMA) at 90/10, 70/30, 50/50, and 30/70 mass fraction % alcohol/monomer before and after evaporation of alcohol. Thin (ca 0.2 mm) disks of human mid-coronal dentin were demineralized and placed in a well beneath the contact probe of a linear variable differential transformer (LVDT). The height of the matrix was measured before and after random application of one of the twelve alcohol/monomer mixtures. Matrix height was measured during infiltration and during solvent evaporation. Between trials, residual monomer was extracted using ethanol. These studies were repeated on specimens in which 100% alcohol was used to substitute for water in the matrix. Both studies revealed that matrices shrink 30-50% but that pretreatment of matrices with alcohol prevents BisGMA phase separations from occurring. Wet bonding with ethanol instead of water permits infiltration of relatively hydrophobic alcohol/monomers.     

Micromechanical analysis of dentin/adhesive interface by the finite element method

The interfacial microstructure and spatial distribution of the modulus of elasticity have a profound effect on load transfer at the dentin/adhesive (d/a) interface. The microstructure is influenced by the varying degree of demineralization of intertubular and peritubular dentin during etching as well as the depth of adhesive penetration into the hybrid layer. These factors lead not only to a unique microstructure in the vicinity of the dentinal tubules, but also to a mechanically graded hybrid layer. This article investigates the micromechanical stress distribution at a d/a interface with the use of finite element analysis (FEA). Such analysis is now feasible given the newly measured moduli of elasticity at micro- and nanoscales. The results indicate that the morphological and micromechanical properties of the d/a interface affects the stress field such that the fracture/failure is likely to initiate in the stress-concentration zone of peritubular dentin next to the hybrid/exposed-collagen layer. The results suggest that devising a full-depth high modulus hybrid layer may considerably reduce the stress concentration zone and the magnitude of stress concentration in the peritubular dentin next to the hybrid/exposed-collagen layer.     

The importance of size-exclusion characteristics of type I collagen in bonding to dentin matrices

The mineral phase of dentin is located primarily within collagen fibrils. During development, bone or dentin collagen fibrils are formed first and then water within the fibril is replaced with apatite crystallites. Mineralized collagen contains very little water. During dentin bonding, acid-etching of mineralized dentin solubilizes the mineral crystallites and replaces them with water. During the infiltration phase of dentin bonding, adhesive comonomers are supposed to replace all of the collagen water with adhesive monomers that are then polymerized into copolymers. The authors of a recently published review suggested that dental monomers were too large to enter and displace water from collagen fibrils. If that were true, the endogenous proteases bound to dentin collagen could be responsible for unimpeded collagen degradation that is responsible for the poor durability of resin–dentin bonds. The current work studied the size–exclusion characteristics of dentin collagen, using a gel-filtration-like column chromatography technique, using dentin powder instead of Sephadex. The elution volumes of test molecules, including adhesive monomers, revealed that adhesive monomers smaller than ～1000 Da can freely diffuse into collagen water, while molecules of 10,000 Da begin to be excluded, and bovine serum albumin (66,000 Da) was fully excluded. These results validate the concept that dental monomers can permeate between collagen molecules during infiltration by etch-and-rinse adhesives in water-saturated matrices.     

Changes in stiffness of resin-infiltrated demineralized dentin after remineralization by a bottom-up biomimetic approach

This study examined changes in elastic modulus, mineral density and ultrastructure of resin-infiltrated dentin after biomimetic remineralization. Sixty demineralized dentin beams were infiltrated with Clearfil Tri-S Bond, One-Step or Prime&Bond NT. They were immersed in simulated body fluid (SBF) for 1 week to maximize water sorption before determining the baseline elastic moduli. For each adhesive (N = 20) half of the beams remained immersed in SBF (control). The rest were immersed in a biomimetic remineralization medium. The elastic moduli were measured weekly for 15 additional weeks. Representative remineralized specimens were evaluated by X-ray microtomography and transmission electron microscopy (TEM). The elastic moduli of control resin-infiltrated dentin remained consistently low, while those immersed in the biomimetic remineralization medium increased by 55–118% after 4 months. X-ray microtomography of the remineralized specimens revealed decreases in mineral density from the beam surface to the beam core that were indicative of external mineral aggregation and internal mineral deposition. Interfibrillar and intrafibrillar remineralization of resin-sparse intertubular dentin were seen under TEM, together with remineralized peritubular dentin. Biomimetic remineralization occurs by diffusion of nanoprecursors and biomimetic analogs in completely demineralized resin-infiltrated dentin and proceeds without the contribution of materials released from a mineralized dentin base.     

Glycosaminoglycan mimetics (RGTA) modulate adult skeletal muscle satellite cell proliferation in vitro

Confocal Raman microspectroscopy (CRM) provides an important and novel means of analyzing the chemical composition of the adhesive/dentin (a/d) interface. The purpose of this study was to develop a method for quantitative determination of the degree of adhesive penetration at the a/d interface using CRM. Three commercial dentin adhesive systems [Scotchbond Multipurpose Plus (SBMP+), Single Bond (SB), and Primer Bond NT (PBNT)] based on the total etch and "wet" bonding technique were examined in this study. Human dentin specimens treated with these adhesives were analyzed with CRM mapping across the a/d interface. Also, Raman spectra were collected on model mixtures of adhesive and type I collagen, and the ratios of the relative intensities of the Raman bands corresponding to adhesive and collagen were used for the construction of calibration curves. By comparing the Raman band ratios of interface specimens to the calibration curves, the percent of adhesive as a function of spatial position across the a/d interface was determined. The results show that there is a gradual decrease in penetration as a function of position for all three adhesive systems while the adhesive concentration gradient decreases in the order of SBMP+ > SB > PBNT. These differences in penetration of the three adhesives at the a/d interface also are discussed relative to the composition and phase segregation in adhesives. Additionally, our results indicate that confocal Raman microspectroscopy is a reliable in situ analytical technique for simple and rapid quantitative determination of adhesive penetration at its interface with prepared dentin.     

A transmission electron microscopy study of mineralization in age-induced transparent dentin

It is known that fractures are more likely to occur in altered teeth, particularly following restoration or endodontic repair; consequently, it is important to understand the structure of altered forms of dentin, the most abundant tissue in the human tooth, in order to better define the increased propensity for such fractures. Transparent (or sclerotic) dentin, wherein the dentinal tubules become occluded with mineral as a natural progressive consequence of aging, is one such altered form. In the present study, high-resolution transmission electron microscopy is used to investigate the effect of aging on the mineral phase of dentin. Such studies revealed that the intertubular mineral crystallites were smaller in transparent dentin, and that the intratubular mineral (larger crystals deposited within the tubules) was chemically similar to the surrounding intertubular mineral. Exit-wave reconstructed lattice-plane images suggested that the intratubular mineral had nanometer-size grains. These observations support a “dissolution and reprecipitation” mechanism for the formation of transparent dentin.     

Application of hydrophobic resin adhesives to acid-etched dentin with an alternative wet bonding technique

Hydrophilic dentin adhesives are prone to water sorption that adversely affects the durability of resin-dentin bonds. This study examined the feasibility of bonding to dentin with hydrophobic resins via the adaptation of electron microscopy tissue processing techniques. Hydrophobic primers were prepared by diluting 2,2-bis[4(2-hydroxy-3-methacryloyloxy-propyloxy)-phenyl] pro- pane/triethyleneglycol dimethacrylate resins with known ethanol concentrations. They were applied to acid-etched moist dentin using an ethanol wet bonding technique that involved: (1) stepwise replacement of water with a series of increasing ethanol concentrations to prevent the demineralized collagen matrix from collapsing; (2) stepwise replacement of the ethanol with different concentrations of hydrophobic primers and subsequently with neat hydrophobic resin. Using the ethanol wet bonding technique, the experimental primer versions with 40, 50, and 75% resin exhibited tensile strengths which were not significantly different from commercially available hydrophilic three-step adhesives that were bonded with water wet bonding technique. The concept of ethanol wet bonding may be explained in terms of solubility parameter theory. This technique is sensitive to water contamination, as depicted by the lower tensile strength results from partial dehydration protocols. The technique has to be further improved by incorporating elements of dentin permeability reduction to avoid water from dentinal tubules contaminating water-free resin blends during bonding.     

Influence of dentinal polyelectrolytes on wet demineralized dentin,a bonding substrate

The objective of this study was to show the influence of dissolved dentinal polyelectrolytes on the characteristics of dentin (bonding substrate) demineralized by citric acid in the absence or presence of ferric chloride. The demineralizing agent was an aqueous mixture of 0, 1, 3, or 10% ferric chloride in 10% citric acid (10-0, 10-1, 10-3, 10-10, respectively). The hypothesis was that the concentration of dissolved dentinal noncollagenous substances, mainly polyelectrolytes soluble in water, must be decreased by their aggregation with ferric ions, which changes the characteristics of demineralized dentin, the rates of demineralization, and dehydration. Cervical bovine dentin was prepared in 3 x 2 x 2-mm blocks, each weighing 20.0 +/- 0.5 mg. The rate of demineralization was investigated by measuring the weight loss resulting from demineralization by immersion in 10 mL of conditioner at 2-h intervals. The dehydration rate of wet demineralized dentin was determined using two methods: (1) weight loss in a desiccator under 263 Pa pressure and (2) differential scanning calorimetry (DSC). Twenty, 12, 8, and 4 h were required to complete demineralization of the blocks with the 10-0, 10-1, 10-3, and 10-10 solutions, respectively. The 10-10 wet demineralized dentin showed the highest rate of dehydration, followed in descending order by the 10-3, 10-1, and 10-0 specimens. Ferric chloride in dentin conditioners provided both a higher rate of dentin demineralization and a higher dehydration rate of wet demineralized dentin. These results suggest that in the presence of ferric chloride, a decreasing amount of dissolved polyelectrolytes aggregated with ferric ions in the substrates may increase the permeability of dentin to water and citric acid. Improvement of monomer permeability is essential to the preparation of good hybridized dentin, providing a more stable and reliable bonding and also protecting the dentin and pulp from infection. A further study of bonding substrates is required in order to understand the role of hybridized dentin in improved dental treatment.     

The effect of stromelysin-1 (MMP-3) on non-collagenous extracellular matrix proteins of demineralized dentin and the adhesive properties of restorative resins

Dentin non-collagenous matrix components (NCPs) are structural proteins involved in the formation, the architecture and the mineralization of the extracellular matrix (ECM). We investigated here how recombinant metalloproteinase stromelysin-1, also termed MMP-3, initiates the release of ECM molecules from artificially demineralized human dentin. Analysis of the supernatants by Western blotting reveals that MMP-3 extracts PGs (decorin, biglycan), and also a series of phosphorylated proteins: dentin sialoprotein (DSP), osteopontin (OPN), bone sialoprotein (BSP) and MEPE, but neither dentin matrix protein-1 (DMP1), another member of the SIBLING family, nor osteocalcin (OC), a non-phosphorylated matrix molecule. After treatment of dentin surfaces by MMP-3, scanning electron microscope (SEM) examination of resin replica shows an increased penetration of the resin into the dentin tubules when compared to surfaces only treated by demineralizing solutions. This preclinical investigation suggests that MMP-3 may be used to improve the adhesive properties of restorative materials.     

Morphological analysis and chemical content of natural dentin carious lesion zones

Dentin is one of the earliest bio-mineralization products to appear in the evolution of vertebrates. Dentin reactions to infection mimic earlier phylogenetic patterns, and carious lesions are divided into different zones which reflect the natural patho-morphological reaction of dentin to the carious attack. It was the aim of this study to investigate deep dentin carious lesions of human molars with combined polarization light microscopy, scanning electron microscopy and energy dispersive X-ray element analysis (EDX) for the determination of different zones of the carious lesions, their extent and the chemical content. Sixteen extracted teeth with deep dentin carious lesions were embedded in Technovit 9100 (Kulzer) and serial sections of 80 microm thickness were made. These sections were then examined with polarized light microscopy to identify the different zones of the lesions. The outlines of the zones were traced consecutively and 3D-reconstructions were made for the determination of the extent and calculation of the volumes of the different zones. From the volumes of the demineralizing dentin and the translucent zone a Dentin Demineralization Index (DDI) was calculated. Three sections of each lesion were then coated with carbon and studied with a scanning electron microscope. 3D-reconstruction of the teeth showed the rather stable translucent zone, interrupted by remnants of dead tracts, and very different volumes of demineralizing dentin. Therefore, with increasing size of the demineralizing dentin, the DDI increased. The chemical content was measured using energy dispersive X-ray analysis (EDX) in areas of intertubular dentin. The content of Ca, P, and C was significantly different in all zones. The Ca/P ratio was significantly different between sound dentin and demineralizing dentin. From the results we conclude that the mineral content of intertubular dentin of the translucent zone and demineralizing dentin is different from that of sound dentin, and the unique mineralization pattern of the translucent zone is a biological reaction to the carious attack. Because active dentin lesions exhibit many non-occluded open dentin tubules, further bacterial invasion or, in case of dentin treatment, the penetration of bonding agents towards the pulp is morphologically not prevented and therefore of clinical importance.     

Interfacial chemistry of class II composite restoration: structure analysis

The gingival margins of class II composite restorations are particularly vulnerable to marginal leakage and secondary caries. In identifying the factors contributing to caries development, the molecular structure and differences in the structure at the proximal and gingival margins have been largely overlooked. The purpose of this study was to compare the molecular structure at the adhesive/dentin interface of the proximal and gingival walls of class II composite restorations. Class II preparations were cut in 12 unerupted third molars with a water-cooled high-speed dental handpiece. The prepared teeth were randomly selected for treatment with Single Bond (SB) + Z100 (3M). Teeth were restored, per manufacturer's directions, under humidity and temperature characteristic of the oral cavity. Restored teeth were kept in sterile Delbecco's phosphate saline for 48 h. The samples were sectioned occluso-gingivally and micro-Raman spectra were acquired at approximately 1.5-microm spatial resolution across the composite/adhesive/dentin interfaces. Samples were wet throughout spectral acquisition. Raman spectral characteristics at the proximal and gingival margins were distinctly different; the depth of demineralized dentin was 6-7 microm at proximal margin, 12-13 microm at gingival margin. SB adhesive penetrated the depth of demineralized dentin in a gradient at the proximal margin. The "single bottle" adhesive used in this study, gradually penetrated the depth of the demineralized dentin at the proximal margin but failed to infiltrate the depth at the gingival margin, leaving a thick exposed collagen layer.     

Chemical profile of adhesive/caries-affected dentin interfaces using Raman microspectroscopy

In clinical practice, dentists must frequently bond adhesives to caries-affected dentin substrates, but the bond that characteristically forms with these substrates does not provide the durability necessary for long-term clinical function. The purpose of this study was to characterize and compare the interfacial chemistry of adhesive with caries-affected and noncarious dentin using micro-Raman spectroscopy. The results indicated that the differences in the Raman spectra between noncarious and caries-affected dentin could not be accounted for by simple decreased mineralization. Both the structure of collagen and mineral in the caries-affected dentin has been altered by the caries process. The differences in structure and composition not only interfered with acid-etching process but also subsequent resin monomer penetration. It was shown that the interface between the adhesive and caries-affected dentin was wider and more complicated than that of the adhesive and noncarious dentin. As a result of adhesive phase separation, a structurally integrated hybrid layer did not form at the interface with either caries-affected or noncarious dentin. Using chemical imaging techniques, this study provides the direct evidence of adhesive phase separation at the interface with caries-affected dentin. Although our group previously reported adhesive phase separation at the interface with noncarious dentin, the chemistry of caries-affected dentin leads to greater variability and a more highly irregular composition along the length and breadth of the interface.     

Enzymatic biodegradation of HEMA/bisGMA adhesives formulated with different water content

Dentin adhesives may undergo phase separation when bonding to wet demineralized dentin. We hypothesized that adhesives exhibiting phase separation will experience enhanced biodegradation of methacrylate ester groups. The objective of this project was to study the effect of enzyme-exposure on the release of methacrylic acid (MAA) and 2-hydroxyethyl methacrylate (HEMA) from adhesives formulated under conditions simulating wet bonding. HEMA/bisGMA(2,2-bis[4(2-hydroxy-3-methacryloyloxy-propyloxy)-phenyl] propane), 45/55 w/w ratio, was formulated with different water content: 0 Wt % (A00), 8 wt % (A08), and 16 wt % (A16). After a three day prewash, adhesive discs were incubated with/without porcine liver esterase (PLE) in phosphate buffer (PB, pH 7.4) at 37 degrees C for 8 days. Supernatants were collected daily and analyzed for MAA and HEMA by HPLC. For all formulations, daily MAA release in the presence of PLE was increased compared to MAA release in PB. HEMA release in the presence of PLE was not detected while HEMA release was consistently measured in PB. A08 and A16 released significantly larger amounts of HEMA compared to A00. Analysis of the cumulative release of analytes showed that the leachables in PLE was significantly increased (p < 0.05) as compared with that released in PB indicating that MAA release was not only formed from unreacted monomers but from pendant groups in the polymer network. However, the levels of analytes HEMA in PB or MAA in PLE were increased in A08 and A16 as compared with A00, which suggests that there could be a greater loss of material in HEMA/bisGMA adhesives that experience phase separation under wet bonding conditions.     

Interfacial chemistry of moisture-aged class II composite restorations

Under in vivo conditions, the adhesive/dentin bond at the gingival margin of class II composite restorations can be the first defense against substances that may penetrate and ultimately undermine the composite restoration. Deterioration of this bond during aqueous aging is an area of intense investigation, but to date, the majority of our techniques have provided only an indirect assessment of the degrading components. The purpose of this study was to analyze the in situ molecular structure of adhesive/dentin interfaces in class II composite restorations, following aging in aqueous solutions. Class II preparations were cut from 12 unerupted human third molars, with a water-cooled, high-speed, dental handpiece. The prepared teeth were randomly selected for restoration with single bond (SB) and Z100 (3M). Teeth were restored, as per the manufacturer's directions, under environmental conditions that simulated humidity and temperature characteristics of the oral cavity. Restored teeth were kept in sterile Delbecco's phosphate saline for 48 h or 90 days. The samples were sectioned occlusogingivally and micro-Raman spectra were acquired at approximately 1.5 microm spatial resolution across the composite/adhesive/dentin interfaces at the gingival margins. Samples were wet throughout spectral acquisition. The relative intensity of bands associated with the adhesive in the interfacial region decreased dramatically after aqueous storage. This decrease in concert with the similar depth of dentin demineralization provides direct spectroscopic evidence of leaching of adhesive monomer from the interface during the 90 days of storage. SB adhesive infiltrated 4-5 microm of 12-microm demineralized dentin at the gingival margin. After 90 days of aqueous storage, SB adhesive infiltration was reduced to approximately 2 microm, leaving approximately 10 microm of demineralized dentin collagen exposed at the gingival margin. The unprotected collagen at the gingival margin of the aged class II composite restorations was disorganized, suggesting hydrolysis of the collagen, with 90 days of aqueous storage.     

Citric acid etching of cervical sclerotic dentin lesions: an AFM study

Atomic force microscopy (AFM) has been used to determine microstructural changes, etching rates of peritubular dentin, and intertubular dentin recession during demineralization in dilute acidic solutions. These studies have not included many forms of altered dentin, including noncarious sclerotic root dentin associated with Cl V (abfraction) lesions. We sought to determine microstructural changes and recession rates during demineralization in citric acid (pH 2.5, 0.018M) for the transparent/sclerotic zone. Highly polished dentin disks were prepared from teeth with noncarious C1 V lesions (n = 3) and compared with normal root dentin (n = 3). Samples were etched at 5-s intervals for 1 min and at longer intervals up to 30 min. The depth changes in various portions of the dentin with respect to the reference layer were measured and changes in microstructure observed in solution in the wet cell of the AFM. In sclerotic dentin, most tubule lumens were occluded with crystalline deposits that etched more slowly than the other dentin components, but etching rates could not be determined due to their irregular geometry. The intertubular dentin recession quickly reached a plateau after a depth change of <1 microm for all dentin types, in agreement with prior work. Mixed linear regression models indicated an important difference between the etching of sclerotic intertubular dentin and that of non-sclerotic root dentin that became apparent after 600 s (p = 0.037). The sclerotic intertubular dentin underwent less depth change at the plateau (558 nm) compared to normal root dentin (744 nm). In addition, normal root dentin underwent significantly greater recession than coronal dentin (p = 0.002). The results of this study indicate that intertubular sclerotic dentin from Cl V lesions etches differently than normal root dentin, and this may help explain the difficulties in restoring such lesions with current bonding procedures.     

Improved wet bonding of methyl methacrylate-tri-n-butylborane resin to dentin etched with ten percent phosphoric acid in the presence of ferric ions

The objective of this study was to determine the influence of dissolved dentinal substances in demineralized dentin on the hybridization of resin for bonding to dentin. It was hypothesized that these substances, including polyelectrolytes, significantly change the substrates, which could then be assessed by the addition of Na(+), Ca(2+), or Fe(3+) in 10% phosphoric acid. Bovine dentin specimens were etched for 10 s with a solution of 10% phosphoric acid (control) or of 22.0 mM dissolved sodium chloride (10P-Na), calcium chloride (10P-Ca), or ferric chloride (10P-Fe). The specimens were then rinsed, blot-dried, and primed three times with 5% 4-methacryloyloxyethyl trimellitate anhydride in acetone for 60 s. Methyl methacrylate-tri-n-butylborane resin was then applied. The tensile bond strength of each of the dumbbell-shaped specimens was then measured. The fractured surfaces and modified cross-sections were examined by scanning electron microscopy. The cross-sections were soaked in 6N HCl for 10 s and then in 1% sodium hypochlorite for 30 min to determine the resin content in the hybridized specimens. Shrinkage of the demineralized dentins upon drying was assessed by atomic force microscopy. The tensile bond strengths were 10.8 +/- 4.5 (control), 15.0 +/- 7.0 (10P-Na), 19.3 +/- 5.5 (10P-Ca), and 27.8 +/- 8.1 (10P-Fe) MPa. The atomic force microscopy studies showed that Fe(3+) minimized the shrinkage by drying for 10 s but Ca(2+) and Na(+) did not decrease the shrinkage the same as the control. The results support the hypothesis that the monomer permeability of wet demineralized dentin is effectively improved by dissolving ferric ions in the phosphoric acid, resulting in a greater bond strength and higher resin content in the hybridized dentin. The dissolved dentinal substances, including the polyelectrolytes, had a significant influence on the characteristics of the demineralized dentin, changing the degree of hybridization and bonding.     

Net expansion of dried demineralized dentin matrix produced by monomer/alcohol saturation and solvent evaporation

The purpose of this work was to determine if nonaqueous methacrylate monomer/alcohol mixtures could expand dried collapsed demineralized dentin matrix. Thin disks (ca. 200 microm) of human dentin were demineralized and placed in wells beneath contact probes of linear variable differential transformers. The probes were placed on water-saturated expanded matrices to record the shrinkage associated with drying. Monomer mixtures containing hydroxyethyl methacrylate, 2,2-bis[4-(2-hydroxy-3 methacryloyloxy)propoxyphenyl] propane, or triethyleneglycol dimethacrylate were mixed with methanol or ethanol at alcohol/monomer mass fraction % of 90/10, 70/30, 50/50, or 30/70. They were randomly applied to the dried matrices to determine the rate and magnitude of expansion; then shrinkage was recorded during evaporation of the alcohols. The results indicated that matrix expansion was positively correlated with the Hoy's solubility parameters for hydrogen bonding forces (delta(h)) of the monomer/solvent mixtures (p < 0.001). Expansions were more rapid with methanol-containing than with ethanol-containing monomer mixtures. For the test solutions, triethyleneglycol dimethacrylate-containing mixtures produced the slowest rate of matrix expansion and hydroxyethyl methacrylate-containing mixtures the most rapid expansion. When the solvents were evaporated, the matrix shrank in proportion to the solvent content and the delta(h) of the monomer-solvent mixtures. The results indicate that expansion of dried, collapsed dentin matrices requires that the delta(h) of the mixtures be larger than 17 (J/cm(3))(1/2). The greater the delta(h) of the monomer solutions, the greater the rate and extent of expansion.     

Analysis of acid-treated dentin smear debris and smear layers using confocal Raman microspectroscopy

Smear layers are generally present on any dentin surface prepared with cutting instruments and are often the only available substrate for bonding. It is commonly reported that acid removes these layers, but to date there has been no chemical evidence to support this observation. Confocal Raman microspectroscopy was used to investigate changes in the composition and molecular structure of acid-treated smear debris and in situ dentin smear layers. The exposed dentin in human molars was abraded with 600-grit silicon carbide sandpaper. Raman spectra were acquired on the smear debris and collected from the sandpaper before and after treatment with 10% citric acid, 35% H3PO4, or 0.5M ethylenediaminetetraacetic acid (EDTA). The resultant smeared dentin samples were treated with one of the aforementioned reagents, and spectra were acquired at 1.0-microm intervals across the interfaces of the smear layers/demineralized dentin/mineralized dentin. Corresponding specimens were morphologically analyzed with scanning electron microscopy (SEM). The results showed that the composition of the smear debris and the in situ smear layers was a mixture of disorganized collagen and mineral. Spectral changes in the smear debris suggested that the disorganized collagen was denatured by acid treatment. The denatured collagen formed a gelatinous matrix around the mineral in the smear layer, thereby shielding it from the acid. The smear layers were not apparent in the SEM micrographs of acid-etched dentin prepared and processed with conventional techniques. The micro-Raman spectroscopic results presented in this study provide the first direct evidence that partially denatured collagen within smear layers is not removed and that the mineral is only partially removed with acids that represent conventional dentin adhesive etchants.