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Effect of dentin primers on the morphology, molecular composition and collagen conformation of acid-demineralized dentin in situ.

OBJECTIVES: The purpose of this study was to assess the effect of two commercial primers and distilled water on the morphology, molecular composition and collagen conformation of acid-demineralized dentin in situ. METHODS: Dentin specimens etched with Scotchbond Etchant were imaged by tapping mode AFM and analyzed by MIR-FTIR spectroscopy. They were then primed with Scotchbond Multi-Purpose Plus Primer, Scotchbond 1 Adhesive or distilled water and imaged and analyzed again. The chemical modifications induced on the uppermost 2 microns of primed dentin were studied after water and original primer subtraction. The conformational changes of type I collagen at this region were evaluated by deconvoluting the amide I band components. The absorbance ratio An(1655/1627) was used to semiquantitatively assess, on a relative basis, the extent of collagen denaturation. RESULTS: All the priming treatments swelled the collapsed dentin collagen left after etching. No evidence of primary bonding was found after priming treatments, while approximately 50% of the conditioned dentin collagen appeared denatured. Treatment with distilled water did not change the status of denatured collagen, however, application of the commercial primers refolded the alpha-helix to approximately 95% of the extent found on the native reference dentin. SIGNIFICANCE: The dynamic response of dentin collagen to demineralization and priming treatments reveals the critical role of some primers in rapidly restoring the conformational status of acid-denatured collagen. Implementation of reactive adhesive groups in alpha-helix recovery may provide an associative means of modifying the mechanical properties of the demineralized collagen based on the extent of their intermolecular bonding.

Acid Etching, Dental↗

Reduced expression of dentin sialophosphoprotein is associated with dysplastic dentin in mice overexpressing transforming growth factor-beta 1 in teeth.

Transforming growth factor (TGF)-beta1 is expressed in developing tooth from the initiation stage through adulthood. Odontoblast-specific expression of TGF-beta1 in the tooth continues throughout life; however, the precise biological functions of this growth factor in the odontoblasts are not clearly understood. Herein, we describe the generation of transgenic mice that overexpress active TGF-beta1 predominantly in the odontoblasts. Teeth of these mice show a significant reduction in the tooth mineralization, defective dentin formation, and a relatively high branching of dentinal tubules. Dentin extracellular matrix components such as type I and III collagens are increased and deposited abnormally in the dental pulp, similar to the hereditary human tooth disorders such as dentin dysplasia and dentinogenesis imperfecta. Calcium, one of the crucial inorganic components of mineralization, is also apparently increased in the transgenic mouse teeth. Most importantly, the expression of dentin sialophosphoprotein (dspp), a candidate gene implicated in dentinogenesis imperfecta II (MIM 125420), is significantly down-regulated in the transgenic teeth. Our results provide in vivo evidence suggesting that TGF-beta1 mediated expression of dspp is crucial for dentin mineralization. These findings also provide for the first time a direct experimental evidence indicating that decreased dspp gene expression along with the other cellular changes in odontoblasts may result in human hereditary dental disorders like dentinogenesis imperfecta II (MIM 125420) and dentin dysplasia (MIM 125400 and 125420).

Animals↗

Dentine hypersensitivity. II. Effects produced by the uptake in vitro of toothpastes onto dentine.

Toothpastes arguably are the most common preparations used in the treatment of dentine hypersensitivity. Evidence for efficacy largely comes from clinical trials, and supports the role of the contained so-called "active" ingredient. How such compounds produce therapeutic benefit has received limited attention. This study measured the availability and uptake onto dentine of fluoride and metal ions contained in commercial and test toothpastes. The effects of exposure of dentine sections to these toothpastes, and abrasive only pastes in particular, was investigated by scanning electron microscopy and X-ray microanalysis. Aqueous extracts of the different toothpastes contained measurable concentrations of the incorporated metal and or fluoride, but usually considerably below that incorporated by the manufacturer. Extraction of dentine sections exposed to toothpaste slurries revealed levels of fluoride and metal ions, although post-treatment washing in water reduced these levels. Surface changes at 1 and 10 h were not consistent with the uptake of fluoride or metals. These changes were almost certainly produced by the contained abrasives, including calcium carbonate, dicalcium phosphate, alumina and silica. In particular fume silica progressively accumulated on the dentine surface to narrow and occlude dentinal tubules. Unlike other abrasives fume silica was resistant to removal by washing. It would appear that insufficient attention has been given to the therapeutic potential of toothpaste ingredients, particularly abrasives, to produce benefit by occlusion of dentinal tubules.

Dentifrices↗

Dentine hypersensitivity. The effects in vitro of acids and dietary substances on root-planed and burred dentine.

Evidence indicates that teeth exhibiting cervical dentine hypersensitivity have open dentinal tubules at the dentine surface. The identification of factors which render dentine exposed and tubules open is important both to the prevention and management of dentine hypersensitivity. In this study, recently extracted teeth were root planed or burred to expose the root dentine. Specimens were horizontally sectioned and then using the apical portion as control, the coronal portions placed in a variety of strong and weak acids and dietary fluids. Examination under the scanning electron microscope revealed a smear layer covering completely underlying tubules on the control root planed or burred portions. Test portions exposed to strong and weak acids showed loss of the smear layer and exposure of large numbers of tubules. Formic and tannic acids produced no changes. Some dietary fluids, in particular red and white wine, citrus fruit juices, apple juice and yogurt produced similar etching effects to the acids. The low pH carbonated drink, coca-cola, and a blackcurrent cordial produced no effects. The results of this study in vitro cannot necessarily be extrapolated to the clinical situation, but suggest that certain dietary factors could play a rôle in the aetiology of dentine hypersensitivity. Dietary advice to patients may prove important in the management of this often recurrent condition.

Acids↗

Transmission electron microscopic appearance of dentin matrix in type II dentin dysplasia.

Dentin matrix of demineralized primary and permanent teeth with type II dentin dysplasia was studied by transmission electron microscopy. The coronal dentin of a maxillary third molar exhibited a normal structure. In the radicular dentin, tubules were few in number; the major part of the dentin was composed of thick, curvy bundles of cross-striated collagen fibers. In the most aberrant areas of the radicular dentin, coarse collagen fibers measuring up to 140 nm in thickness were observed. The dentin of the primary tooth showed a similar, but somewhat less irregular, structure.

Collagen↗

Refined mapping of the human dentin sialophosphoprotein (DSPP) gene within the critical dentinogenesis imperfecta type II and dentin dysplasia type II loci.

Dentinogenesis imperfecta type II and dentin dysplasia type II are diseases resulting in abnormal dentin formation, which have been mapped to overlapping regions of human chromosome 4q defined by markers D4S2691 and D4S2692 (6.6 cM) and D4S3291 and SPP1 (14.1 cM), respectively. Recently, two of the major non-collagenous proteins of dentin, dentin sialoprotein (DSP) and dentin phosphoprotein (DPP, phosphophoryn) have been shown to be encoded by a single gene, termed dentin sialophosphoprotein (DSPP), which has been mapped to human chromosome 4. The purpose of this study was to perform refined mapping of DSPP related to these disease loci by gene content mapping, as well as to place the DSPP gene on the physical map of human chromosome 4 by sequence tagged site (STS) content mapping. Human genomic DSPP clones were isolated, and gene content mapping performed with specific primers for dentin matrix protein 1 (DMP1), bone sialoprotein (BSP) and osteopontin (secreted phosphoprotein 1, SPP1). STS content mapping was then performed with flanking STS markers to these dentin/bone gene loci. Our results demonstrate that the DSPP and DMP1 genes are within a maximum distance of 110 kb. Both DSPP and DMP-1 have been placed on the physical map of human chromosome 4 within the interval defined by markers D4S564 and D4S1292. DSPP is thereby strengthened as a candidate gene for both DGI-II and DD-II.

Amino Acid Sequence↗

Dentine hypersensitivity: the measurement in vitro of streaming potentials with fluid flow across dentine and hydroxyapatite.

Stimulus transmission across dentine, in conditions such as dentine hypersensitivity, is considered to occur via a hydrodynamic mechanism. This fluid flow in dentine may then induce a mechanoreceptor response in pulpal nerves. However, when fluid flows through a porous structure electrical potentials are also generated. The aim of this study was to develop a reproducible model system to measure streaming potential across dentine and hydroxyapatite and determine the influence of pressure. Using an acrylic cell, with silver electrodes, streaming potentials were recorded across dimensionally standardized dentine and hydroxyapatite specimens, over a pressure range of 1-6 atmospheres. Streaming potentials were found to be directly proportional to pressure and dependent on the electrical conductivity of the saline used in the cell. The results confirm the limited existing data on streaming potentials across dentine and indicate that at these low pressures excitation of pulpal nerves would not occur. However, if, as may be the case, stimuli applied to dentine create very high pressures, the resultant potentials generated could indeed evoke a neural response. The model system is worthy of further use to study this phenomenon and the factors which may influence it.

Dentin↗

Dentin hypersensitivity. Occlusion of dentinal tubules by brushing with and without an abrasive dentifrice.

Daily toothbrushing with and without an abrasive dentifrice was performed on the ground surfaces of human cervical dentin attached to resin plates, which had been exposed to the oral cavities of 4 subjects for 8 weeks. Brushing with a dentifrice containing calcium hydrogen phosphate as an abrasive caused most of the dentinal tubules to open. The patency of dentinal tubules is thought to induce dentin hypersensitivity. Brushing without a dentifrice, on the other hand, resulted in occlusion of the dentinal tubules with organic pellicle-containing minerals. Furthermore, abrasion did not occur; however, no bacterial plaque or dental calculus were retained. The occluded tubules which were distributed over the entire dentin surfaces will likely prevent hypersensitivity. Such organic-mineral materials occluding the tubules can be derived from saliva and embedded by the brushing. We suggest that use of a non-abrasive dentifrice will prevent or reduce dentin hypersensitivity in the cervical regions.

Adult↗

Effect of functional monomer in commercial dentin bonding agents use of an experimental dentin bonding system.

The objective of the present study was to understand the role of the functional monomers in dentin bonding agents of an experimental dentin bonding system by measuring the wall-to-wall contraction gap and tensile bond strength measurement. The efficacy of three commercial dentin bonding agents after using EDTA for conditioning and GM for priming was evaluated by measuring the contraction gap of the resin composite in a cylindrical dentin cavity, and by measuring the tensile bond strength of the composite to a flat dentin surface. The effect of the functional monomers was demonstrated by the contraction gap measurement alone. The value of the contraction gap was significantly different between the commercial dentin bonding agents and these agents without functional monomers (p < 0.05). It was concluded that the functional monomers were essential to obtaining the marginal integrity of the resin composite in the dentin cavities.

Acrylic Resins↗

Effect of primer solvent, primer agitation, and dentin dryness on shear bond strength to dentin.

PURPOSE: To evaluate the shear bond strength of acetone- or water-based primers applied with or without agitation to either wet or dry dentin. MATERIALS AND METHODS: Forty-eight caries-free extracted human premolars were embedded in gypsum and flat dentin surfaces were prepared. The teeth were randomly divided in eight groups with six specimens each. After acid etching and rinsing, either a HEMA and acetone (Ac) or a HEMA and water (Wa) primer were applied to dry (D) or wet (W) dentin. The primer was placed with (Y) or without (N) agitation according to the following combinations: Group 1: D-Ac-Y; Group 2: D-Ac-N; Group 3: D-Wa-Y; Group 4: D-Wa-N; Group 5: W-Ac-Y; Group 6: W-Ac-N; Group 7: W-Wa-Y; Group 8: W-Wa-N. The primed surface was dried, covered with a bonding resin and light-cured, whereupon a composite cylinder was bonded to the dentin surface. The shear bond strength was determined after 30 days in water. RESULTS: The mean and standard deviations, expressed in MPa, were: Group 1: 13.1 +/- 3.42; Group 2: 15.9 +/- 4.84; Group 3: 19.8 +/- 6.64; Group 4: 18.6 +/- 5.18; Group 5: 16.5 +/- 5.25; Group 6: 25.5 +/- 4.79; Group 7: 21.9 +/- 4.94; Group 8: 17.4 +/- 4.50. The results were analyzed using ANOVA and Waller-Duncans K-ratio t-test. The acetone-based primer gave the highest bond strength to wet dentin without agitation. When the acetone-based primer was used on dry dentin a significant decrease in bond strength occurred (P < 0.05). No significant difference (P > 0.05) was found for the water-based primer on wet or dry dentin. Agitation improved the bond strength for the water-based primer, but decreased the bond strength for the acetone-based primer.

Acetone↗

The influence of tubule density and area of solid dentin on bond strength of two adhesive systems to dentin.

PURPOSE: To determine the correlation between the tubule density (TD) and the area occupied by solid dentin (ASD) with the bond strength of one conventional and one self-etching adhesive system to dentin. MATERIALS AND METHODS: The crown of extracted human third molars was transversally sectioned with a diamond saw to expose either superficial, middle, or deep dentin. The three groups of dentin surfaces were randomly divided and bonded with either Clearfil Liner Bond 2V (LB) or Prime & Bond 2.1 (PB) adhesive systems according to manufacturer's directions. Resin composite buildup crowns (10.0 mm high) were incrementally constructed on the bonded surfaces and the teeth stored in water at 37 degrees C. After 24 h of storage, the teeth were vertically, serially sectioned in both x and y directions to obtain several bonded sticks of approximately 0.7 mm2 cross-sectional area. Each stick was tested in tension in a EMIC DL-500 tester at 0.5 mm/min until failure. After testing, the dentin side of the fractured specimen was gently abraded with a 1000-grit SiC paper, etched with 37% phosphoric acid for 15 s and allowed to air dry. SEM micrographs at 1000x and 4000x magnification were taken to permit calculation of the TD (number of tubules/mm2) and ASD (% of total area) at the site of fracture. Correlation between TD and ASD with the bond strength data was performed by linear regression. All statistical analysis was done with alpha = 0.05. RESULTS: Overall bond strength (MPa) for LB was 26.0 +/- 10.2, and 42.6 +/- 15.2 for PB. There was a significant direct relationship between bond strength and ASD for both materials (r2 = 0.20, p < 0.05 and r2 = 0.66, p < 0.01, respectively for LB and PB). PB bond strength dropped significantly as the TD increased (r2 = 0.63, p < 0.05), while LB was not sensitive to TD (r2 = 0.05, p > 0.05). Mean bond strength of PB was significantly higher than LB for both superficial and middle dentin (p < 0.05), while there was no significant difference for deep dentin (p > 0.05). CONCLUSION: Regional variations in TD and ASD may modify bond strength of both conventional and self-etching adhesive systems. Bonding sites with larger ASD seem to yield higher bond strengths regardless of the type of adhesive system used.

Acetone↗

Effect of bur cutting patterns and dentin bonding agents on dentin permeability in a fluid flow model.

This study investigated the effects of bur cutting surface roughness and bonding systems on dentin permeability. A conventional straight edged bur, cross-cut serrated bur and an extensively serrated bur were utilized with two different bonding systems. Null hypothesis was that increased surface roughness does not decrease the permeability of dentin sealing after application of bonding agents. This study incorporated a fluid flow model for measuring dentin permeability. Seventy caries-free extracted molars were used and sectioned 2 mm apical to the cementoenamel junction (CEJ). The pulp tissue was removed and the chamber cleaned using 37% phosphoric acid followed by 1:1 sodium hypochlorite solution. The specimens were mounted to plexiglass plates using cyanoacrylate cement. A flow system was established through the pulp chamber and gravity pressurized phosphate buffered saline (PBS) was forced into the chamber and the "closed system" permeability measured. Burs were used to prepare flat surfaces in dentin, and after etching with phosphoric acid, the "open tubule" permeability was determined. After applying the dentin bonding agents, fluid flow from the pulp chamber into dentin was again measured. The percentage of reduction in permeability following bonding agent application was then calculated. A two-way ANOVA found a significant (p<0.05) decrease in bonded dentin permeability in specimens prepared using the cross-cut serrated and extensively serrated bur as compared to the conventional straight-edged bur. No significant differences were detected between bonding systems and no combination of surface topography or bonding agent completely sealed the surface.

Analysis of Variance↗

Scanning electron microscopy evaluation of the link between a new dentinal desensitizer and dentine.

AIM: The purpose of this study was to evaluate the interaction of a novel dentinal desensitizer Twin Pro with enamel-dentinal adhesive preparations or filling materials and dentine using scanning electron microscopy (SEM) technology. METHODS: Black's I class cavities were drilled extracted molar teeth free of caries or fissures, and the cavities disinfected. The specimens were divided into 4 groups of 2 teeth each treated as follows: Group A: Twin Pro, fluid (Tetric-Flow Vivadent) and micro-hybrid (Tetric-Ceram Vivadent) composites placed on the etched (Liner Bond 2V Clearfil) cavities. Group B: Twin Pro, PQ1-Ultradent loaded Primer-Bonding plus fluid and micro-hybrid composites applied to the acid-etched (37% orthophosphoric acid) cavities. Group C: Twin Pro plus Silver amalgam alloy (Phasealloy- Sybram Kerr). Group D (control group): self-etching primer plus fluid and micro-hybrid composite. Specimens were investigated by SEM. RESULTS: The results obtained in all groups show that the application of Twin Pro does not alter the adhesiveness of the restorative composite materials to the dental wall. In fact the desensitizer and the adhesive layers are indistinguishable at SEM observation, and well adherent to the dentinal surface of the cavities. CONCLUSION: The results of SEM investigations show that Twin Pro does not decrease the adhesion of restorative materials to the tooth surface, as it establishes an efficient interconnection with the different materials used, and it forms a uniform layer covering and occluding dentine tubules, and this might constitute an efficacious sealing of dentinal tubules which possibly contribute to decrease dentinal sensitivity to environmental nociceptive stimuli.

Acrylic Resins↗

Influence of metal conditioner contamination of the dentin surface on bond strengths of dentin adhesive systems using self-etching primers.

Carious lesions around crown margins sometimes lead to failure of fixed prosthodontics. This study examined the influence of metal conditioner application on a dentin surface prior to bonding procedures, using two-step self-etching primer systems. Commercially available metal conditioners and self-etching primer dentin bonding systems were used. Bovine mandibular incisors were mounted in self-curing resin, and the facial dentin surfaces were ground wet on 600-grit SiC paper. The metal primers were applied on the dentin surface followed by bonding procedures with four different types of self-etching primer systems. The resin composites were condensed into a mold on the dentin surface and light activated. Ten specimens per test group were stored in 37 degrees C water for 24 hours; they were then shear tested at a crosshead speed of 1.0 mm/minute. Two-way ANOVA and Tukey's HSD tests were done. When the metal conditioners were applied on dentin surfaces before bonding procedures, there was a tendency for decreased dentin bond strengths compared to those obtained with the controls. This tendency differed among the combinations of metal conditioners and self-etching primer systems used. Appropriate surface treatments are required to get optimum bond strengths with the use of technique sensitive bonding systems combined with metal conditioners.

Analysis of Variance↗

Dentine permeability and dentine adhesion.

OBJECTIVES: The objectives of this paper are to review the structure of dentine as it pertains to adhesive bonding and to describe the importance of resin permeation into dentinal tubules and into spaces created between collagen fibrils by acid-etching during resin bonding. The advantages and disadvantages of separate acid-etching, priming and adhesive applications are discussed. DATA SOURCES: Although not an exhaustive review, the concepts included in the review were obtained from the dentine bonding literature. STUDY SELECTION: Attempts were made to critically evaluate what is known about dentine permeability and adhesion and what remains to be discovered. Speculations were made on a number of controversial issues that are not yet resolved. CONCLUSIONS: Acid-etching of dentine produces profound changes in the chemical composition and physical properties of the matrix which can influence the quality of resin-dentine bonds, their strength and perhaps their durability.

Acid Etching, Dental↗

A longitudinal study of dentine lead levels and intelligence, school performance and behaviour: the measurement of dentine lead.

Shed deciduous teeth were obtained from a sample of 996 New Zealand children who were participants in a longitudinal study. For each child, estimates of dentine lead were obtained from two samples of dentine. The information was used to estimate the magnitude of random errors of measurement in dentine lead values and it was estimated that between 15 and 20% of the variance in dentine lead values was attributable to random errors of measurement. In addition, dentine lead levels were influenced by small systematic errors of measurement arising from the age at which the tooth was collected and the type of tooth analysed. These sampling factors, however, accounted for less that 3% of the variance in dentine lead values. The implications of these findings for the interpretation of correlations between dentine lead levels and other variables are discussed.

Child↗

Effect of a chemo-mechanical caries removal system (Carisolv) on dentin topography of non-carious dentin.

The purpose of the present study was to examine the morphology of healthy dentin surfaces after treatment with Carisolv followed by conditioning with phosphoric acid and EDTA, since surface morphology may be of interest for dentin bonding. Another purpose was to evaluate the effect of treatment with Carisolv on healthy non-carious dentin surfaces with exposed collagen fibers. Scanning electron microscopy was utilized to carry out a detailed morphological examination of the dentin surfaces with regard to presence or absence of both smear layer and collagen fibers. Twelve premolars extracted for orthodontic reasons from young adults were used. The two etchants appeared to have produced two distinctly different surfaces. Etching with phosphoric acid following Carisolv treatment resulted in a porous dentin surface, while EDTA etching without prior Carisolv treatment appeared to have uncovered an intact collagen network. In contrast, the surfaces treated with Carisolv prior to EDTA etching displayed smooth intertubular surfaces with only occasional fibers. Apparently, the ability of EDTA to expose collagen in the dentin surface is counteracted or inhibited by the Carisolv treatment. Furthermore, it cannot be excluded that the Carisolv treatment in itself may have dissolved collagen fibers. Since most bonding systems claim bonding to the collagenous component of dentin, the question arises which of the etched surfaces is preferable and to what degree the collagenous component contributes to bonding strength. Further studies are thus needed to evaluate the micromechanical retention of a restoration to the different surfaces described in the present study.

Acid Etching, Dental↗

Phosphoprotein analysis of sequential extracts of human dentin and the determination of the subsequent remineralization potential of these dentin matrices.

Phosphoprotein appears to play an important role in the mineralization of dentin during tooth development and remineralization after demineralization by dental caries. To better understand this role, we describe the extraction and characterization of phosphoprotein from immature, human root apex dentin during and after EDTA demineralization. The extraction procedure included dissociation of the demineralized dentin matrix by guanidine hydrochloride (Gdn.HCl) followed by subsequent digestion with cyanogen bromide (CNBr) and collagenase. Characterization of these extracts included 'Stains-All' staining of SDS polyacrylamide gels (SDS-PAGE) and amino acid, protein and phosphorus analyses. The ability of these matrices to remineralize was determined by TEM and measuring calcium levels in the remineralized tissue by atomic absorption spectroscopy. The staining of SDS-PAGE gels and amino acid analysis showed that an intact phosphophoryn was extracted from the dentin of the immature apices during EDTA demineralization and that it had an apparent Mr approximately 140,000. In the subsequent extracts and digests, the phosphoprotein has a range of molecular weights, some of which may have been degraded products of the intact phosphoprotein. A greater quantity of phosphoprotein was found in the EDTA-demineralized dentin matrices than in dentin after Gdn.HCl, CNBr and collagenase digests. These EDTA-demineralized matrices also remineralized to a greater extent than those dissociated with Gdn.HCl. The differences in both the quantity and the quality, as defined by the amino acid residue profile, of the phosphoprotein in the sequential extracts of the root apex dentin may be important in affecting the ability of this tissue to remineralize.

Adolescent↗