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Dentin sialoprotein isoforms: detection and characterization of a high molecular weight dentin sialoprotein.

Dentin sialoprotein (DSP) is a glycoprotein accounting for 5-8% of the dentin non-collagenous proteins. The cDNA sequence predicts that rat DSP has 13 potential casein kinase phosphorylation sites and six potential N-linked glycosylation sites. However, its total phosphorylation level, as well as the nature and locations of the carbohydrate moieties, are unknown. Our findings in the present study show that rat DSP has 6.2 phosphates per molecule and that the majority of carbohydrates are attached to the protein through N-linked glycosylations. During our separation of dentin non-collagenous proteins with ion-exchange chromatography, we observed high molecular weight components eluting late in the salt gradient that were recognized by anti-DSP antibodies. We have purified these high molecular weight components using a monoclonal anti-DSP antibody affinity column. Data from amino acid analysis, phosphate level measurements and Edman degradation of tryptic peptides unequivocally proved that the very acidic, high molecular weight components are isoforms of DSP (designated HMW-DSP). Deglycosylation analysis indicates that the slower migration rate of HMW-DSP on SDS-PAGE results from its higher level of carbohydrate modifications.

Amino Acid Sequence↗

Evaluations of the interface between amalgam and dentin with a single component dentin bonding system.

The purpose of this scanning electron microscope study was to observe the dentin-amalgam interface after utilizing a standard restorative liner and a single-component dentin adhesive. Continuous interface gaps of over 15 mu were observed in specimens containing no lining and Copal varnish. Discontinuous 5 microns gaps were observed in specimens treated with PQ1. Additionally, specimens treated with PQ1 exhibited resin interpenetration into the dentin surfaces.

Adhesives↗

Self-contamination of deep dentin by dentin fluid.

PURPOSE: To compare, with the use of a resin replica technique, surface features of deep, acid-conditioned dentin from vital human molars that were anesthetized with an anesthetic: Group 1: without a vasoconstrictor (Mepivacaine 3%), and Group 2: containing a vasoconstrictor (Lidocaine 2% with 1:80,000 epinephrine). MATERIALS AND METHODS: 10 Class I cavities with dentin caries were included in each group. Following complete caries removal, a total-etch technique was performed with 32% phosphoric acid (Uni-Etch) for 15 s. After rinsing, each cavity was air-dried for 1 s, then a slow setting vinyl polysiloxane (President) impression was taken. As a control, impressions were taken from three additional cavities in teeth that were anesthetized with Mepivacaine 3% but not acid etched. A TEM-grade epoxy resin was used to prepare replicas from the impressions. Polymerized replicas were coated with gold and examined with a scanning electron microscope. RESULTS: In Group 1, fluid was visible leaving tubular orifices in all replicas. In addition, three were covered with a smooth, amorphous film that was different from the granular appearance of the smear layer in the control cavities. In Group 2, patent tubular orifices without fluid were observed in all specimens. Odontoblast process-like structures were seen from some dentin tubules.

Acid Etching, Dental↗

Resin-dentin interfacial ultrastructure and microtensile dentin bond strength after five-year water storage.

OBJECTIVE: To evaluate a total-etch three-step adhesive system's resin-dentin interfacial ultrastructure and microtensile dentin bond strength (microTBS) after multi-year storage in water. METHODS: Resin composite crowns were formed on 600 grit SiC flattened extracted human molars using a total-etch three-step adhesive system (Optibond FL, Kerr) and a hybrid resin composite (Prodigy, Kerr). microTBS specimens were fabricated and placed in water with 0.5% chloramine T at 37 degrees C until respective static load to failure testing at one-month, six-months and five-year storage. Failure modes were determined by scanning electron microscopy. The interfacial ultrastructure of the resin-dentin interface was analyzed by transmission electron microscopy (TEM) at 48-hours and 44-months storage. microTBS was modeled with Weibull distribution for survival analysis and failure curve distributions were analyzed by the Wald chi-square statistic for significant differences at alpha=0.05. RESULTS: The characteristic tensile strength (sigma omicron) at one-month, six-months and five-year storage was 52.63, 14.77 and 23.57 Mpa, with a Weibull modulus of 3.04, 1.56 and 1.28, respectively. Failure distributions for all groups were significantly different (p<0.0001) with one-month > five-year > six-months. TEM interfacial morphology demonstrated hydrolytic degradation of hybrid layer components at 44-months storage. SIGNIFICANCE: The decrease in tensile strength and changes in ultrastructure may be caused by water sorption and resultant hydrolytic degradation of the adhesive joint.

Absorption↗

Effect of dentin adhesives used as sealers and provisional cementation on bond strength of a resin cement to dentin.

PURPOSE: To evaluate the effects of dentin adhesives employed as resin sealers and provisional cementation on the bond strengths of a resin cement to dentin. METHODS: A two-step etch-and-rinse adhesive (Excite DSC--Group 1) and two-step self-etch adhesive (AdheSE--Group 2) were applied to exposed dentin surfaces prepared from human molars (N=4). Water was used instead of a resin sealer in control Groups 3 and 4. A eugenol-free provisional cement (except for Group 4) was applied to the treated surfaces. After storing in distilled water for 1 week, the provisional cement was removed and cylindrical composite blocks were luted with a resin cement (Variolink II). 0.9 x 0.9 mm sticks were produced from these luted specimens for microtensile bond testing and SEM examination. RESULTS: One-way ANOVA revealed that neither the resin sealer nor the temporary eugenol-free cement had a negative effect on the final bond strength (P> 0.05). Mixed failures were predominantly identified from SEM.

Acrylic Resins↗

Effect of age of sample on the shear bond strength of dentin bonding agents to dentin.

The effect of sample age on the shear bond strengths of two dentin bonding agents to human dentin was evaluated by bonding composite resins to human dentin treated with either Imperva-Bond or Prisma Universal Bond 3. One-half of the specimens were tested at 5 minutes, while the remainder were thermocycled and tested at 2 weeks. Results were analyzed using two sample t tests at the 0.05 probability levels. Failures were evaluated using light microscopy and categorized as adhesive, cohesive within resin, cohesive within tooth, or mixed. The bond strengths were significantly higher for Imperva-Bond than for Prisma Universal Bond 3 at both time periods. For both bonding agents, the 2 week bond strengths were significantly higher than the 5 minute bond strengths. The 2 week bond strength of Imperva-Bond approaches that of unfilled resin to etched enamel.

Adhesives↗

Effect of a carious dentin softener on the dentin and pulp.

The application of GK-101 to carious dentin softens the first or outer layer which contains denatured collagen. The second or inner layer of carious dentin, containing normal collagen fibers, is unaffected. The pulps of dog teeth were not affected by applying the solution to freshly prepared sound dentin.

Animals↗

The effect of dentin pre-treatment and heat-augmented cure on marginal gap formation of a dentin bonding agent.

This study evaluated the effect of enhancing the cure of a DBA and the effect of dentin surface pre-treatment on marginal gap formation. Dentin cavities were prepared on extracted teeth and were pre-treated either with EDTA or phosphoric acid. Marginal leakage was expressed as percent length of margins with gaps. The mean percent length of margins with gaps for the chemically cured groups ranged from a maximum of 97% to a minimum of 85%. For the chemical/thermal-cured group, the mean percent length of margins with gaps was 95%, 95%, 73%, 35%, and 65%, corresponding to the following: pretreatment, no pre-treatment, EDTA, and phosphoric acid for 60, 15, or five s, respectively. SEM micrographs showed partial removal of one smear layer with EDTA and total removal of the smear layers with phosphoric acid. It is concluded that dentin pre-treatment methods have an effect on the percent length of margins with gaps when the DBA used had its cure supplemented with heat.

Acid Etching, Dental↗

The effects of luting resin bond to dentin on the strength of dentin supported by indirect resin composite.

OBJECTIVES: The purpose of this study was to evaluate the effects of thickness and adhesion of three resin cements on the fracture resistance of indirect resin composite bonded to dentin. METHODS: A disk of resin composite used for indirect restorations was bonded to a disk of bovine dentin using three kinds of resin cements with various bonding procedures. The bonding procedures were planned into five groups according to the materials and methods, and subsequently subdivided into three groups according to the cement thickness (50, 150 and 500 microm) in each bonding procedure. The thickness of the resin cement and that of the resin composite disk was changed simultaneously while maintaining a total specimen thickness of 2 mm. The prepared specimens were then stored in water at 37 degrees C for 24h at which time they were trimmed to a size of 2 x 2 x 8 mm. The trimmed specimens were subjected to a three point bending test and the fracture load determined. The tensile bond strength of each bonding procedure was measured and the correlation to the fracture load evaluated. RESULTS: The fracture load was affected by the dentin bond strength. The effect of cement film thickness on the fracture load was negligible. SIGNIFICANCE: When an indirect restoration is adhered to the tooth substrate, the adhesion of the luting cement to the tooth substrate is very important for the fracture resistance of indirect resin composite.

Acid Etching, Dental↗

Dentin sialoprotein, dentin phosphoprotein, enamelysin and ameloblastin: tooth-specific molecules that are distinctively expressed during murine dental differentiation.

Dentin sialophosphoprotein [designated DSPP and cleaved into dentin sialoprotein (DSP) and dentin phosphoprotein (DPP)], enamelysin and ameloblastin are each expressed in unique fashions during tooth development. It is possible that these components participate in cell differentiation and the conversion of unmineralized matrix into mineralized structures. In order to delineate the timing and the positioning of these three molecules in a physiological context, we compared their expression profiles by performing in situ hybridization experiments on consecutive sections in developing mouse tissues. Hybridization signals were uniquely detected for DSPP mRNA in odontoblasts and preameloblasts, for enamelysin mRNA in odontoblasts and in the facing ameloblast layer, and for ameloblastin mRNA in preodontoblasts, polarizing odontoblasts and ameloblasts. Immunohistochemistry showed that DSP and ameloblastin transcripts were translated into proteins that were deposited at the apical pole of the differentiated cells (odontoblasts and ameloblasts, respectively). The interrelated expression profiles found for these tooth-specific molecules illustrate the importance of a specific molecular network to initiate highly regulated processes such as cytodifferentiation and the subsequent mineralization.

Ameloblasts↗

A longitudinal study of dentine lead levels, intelligence, school performance and behaviour. Part III. Dentine lead levels and attention/activity.

The relationship between dentine lead levels and maternal/teacher ratings of inattentive/restless behaviour in children was examined for a birth cohort of New Zealand children. There were small but relatively consistent and stable correlations between dentine lead values and behaviour ratings. After correction for errors of measurement in dentine lead values and behaviour ratings it was estimated that the correlation between lead levels and inattentive/restless behaviour was in the region of +0.18. However, after control for various sources of confounding there was only a small, but statistically significant, correlation of +0.08 between lead levels and inattention/restless in children. It is concluded that the weight of the evidence favours the view that there is a very weak causal association between lead levels and attention and activity levels in children.

Achievement↗

Rat dentin matrix protein 3 is a compound protein of rat dentin sialoprotein and phosphophoryn.

During cloning of the rat incisor phosphophoryn gene, several clones were identified with a PP antibody. One of the clones (2PP) was shown to encode a PP denoted as dentin matrix protein 2, DMP2. We have now sequenced another clone, which appears to be bifunctional, or at minimum, has two distinct domains. The 5' region encodes for dentin sialoprotein (DSP), while the 3' region encodes a small or "mini" PP. There is no stop codon between these domains. The whole gene has been named Dmp3, in keeping with the current nomenclature adopted in our laboratory. The previously reported Dmp2 gene does not encode a DSP domain but has a 114 amino acid carboxy terminal sequence identical except for a single residue with that of the DMP3. Thus, Dmp2, Dmp3 and their corresponding proteins, probably represent related members of a multigene family. There is no evidence for differential splicing. Since the DSP isolated from dentin does not carry the mini-PP domain, it must be cleaved postranslationally from DMP3.

Amino Acid Sequence↗

Studies on adhesion to tooth substrate. 6. Adhesion of composite resin to dentin mediated by the dentin primer containing MTYA.

The influences of the kinds and concentrations of the aldehydes in the primer on dentin adhesion were investigated. Moreover, the durability of the adhesion to the dentin treated with primer was examined by thermal cycling tests. Among the aldehyde tested here, glutaraldehyde was effective in improving the adhesion to dentin etched with citric acid. When HEMA was substituted with ethanol, the bond strengths decreased. The relationship between the bond strength and the concentration of glutaraldehyde in the primer was studied. In the cases of phosphoric acid or citric acid etching, the bond strengths were affected by the concentration of glutaraldehyde in the primer. The bond strengths decreased after 2000 thermocycles in phosphoric acid etching.

Adhesiveness↗

[Dentin-dentinal adhesive interaction. Calorimetric measurement].

Among the tests employed to evaluate the bond strength between dentine and dental adhesives (tensile or shear bond strength measurements, marginal leakage measurements) a general accordance on their reliability cannot be found. Calorimetric measurement may be useful in deciding if the adhesion is mechanical or chemical. In this preliminary study, calorimetric measurements have been performed on three adhesive systems commonly employed: Scotchbond, Gluma, Dentin Adhesit. A calorimeter, equipped with reaction vessels which allow the contact between the two reactants after the thermostatisation has been reached, was used. A thermal effect was observed in all cases, greater for Scotchbond and Gluma, poor for Dentin Adhesit. These values are the sum of different contributes of various reaction paths.

Acrylic Resins↗

[The properties of dentin and resinified dentin in the tensile test].

The purpose of this study is to evaluate the effect of resinifying therapy to the hard tissue of tooth by using measurement of the tensile strength. The fresh extracted maxillary anterior teeth having been cleaned and sterilized were divided into two groups. The control group was placed into 9% N-saline solution of 37 +/- 1 degrees C. The experimental group was treated with resinifying therapy. The tensile specimens were shaped by turning in a lathe until it satisfied the tensile test requirement of Mechanics of Materials except that they were hollow. A set of special split jig was made for this test. Instron Universal Testing Machine was used for measuring the load on the specimens. The loading speed was 0.05 mm/min. The straining meter was used to measure the tensile strain. The result revealed that tensile strength of resinifying dentin was little lower than that of unresinified dentin. The resinified dentin didn't increases stiffness or brittleness.

Acrylic Resins↗

The effect of airborne particle abrasion on the dentin smear layer and dentin: an in vitro investigation.

This study investigated the effect of airborne particle abrasion on the exposed dentin surfaces of 40 extracted teeth. A Siroflow abrasion unit was used to treat four groups of ten specimens that were subjected to 5, 10, 20, and 40 seconds of abrasion respectively. Statistical analysis (Spearman Rank Order) showed a significant difference between all groups (P < .05) but not within groups (P > .05). Scanning electron micrographs showed smear layer removal as an immediate effect of air polishing. Application times of longer than 5 seconds showed obstruction of dentinal tubule openings, possibly a result of abrasive powder residue. It was concluded that airborne particle abrasion removes the smear layer, and the amount of dentin removed corresponded to the time of application.

Air Pressure↗

Eugenol diffusion through dentin related to dentin hydraulic conductance.

OBJECTIVES: The purpose of this study was (1) to find an easy way of evaluating the concentration of eugenol in cell culture fluids; (2) to confirm the relationship between the concentration and the cytotoxicity of eugenol in vitro; (3) to evaluate the cytotoxicity of four temporary eugenol-based filling materials: IRM, super EBA, Kalsogen and zinc oxide-eugenol cement; and (4) to establish a relationship between dentin permeability, eugenol diffusion and cytotoxicity. METHODS: (1) The concentration of eugenol was measured with a spectrofluorimeter; (2) the cell viability of L 929 cells cultivated for 24 h with eugenol-containing medium was evaluated by the MTT assay; (3) after measurement of hydraulic conductance, occlusal cavities in human teeth in vitro were filled with the restorative materials. The cytotoxicity was measured with undiluted test medium and with various dilutions in culture medium; (4) after Lp measurement, the eugenol concentration in the media in the pulp chamber that diffused from IRM and 10(3) mol/l eugenol solution was measured. RESULTS: (1) A proportional relationship (p = 0.001 and r = 1) was found between the concentration of eugenol; (2) eugenol started to be cytotoxic at 10(-5) mol/l and killed 95% of the cells at 10(-3) mol/l; (3) zinc oxide-eugenol cement was the most cytotoxic filling material when tested with the 1:100 dilution; (4) a significant relationship was found between Lp and cytotoxicity (p = 0.04) depending on the dilution of the test medium. A significant relationship was found between Lp and eugenol diffusion from a 10(-3) mol/l solution (p = 0.03) but not between Lp and eugenol diffusing from solid IRM (non significant). SIGNIFICANCE: Eugenol diffusion from zinc oxide-eugenol cement appears to depend more on the role of hydrolysis of eugenol from zinc oxide-eugenol cement than on dentin permeability.

Analysis of Variance↗