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PubMed1980

Phosphoprotein modulation of apatite crystallization.

Several phosphoprotein preparations (phosvitin, rat incisor and fetal calf molar dentin phosphoproteins) all inhibit apatite growth/replication from pre-existing crystal seeds in metastable solutions. Two stages of the crystal growth process were inhibited by these phosphoproteins. First an initial lag period was induced, probably associated with seed surface phenomena. This period was prolonged indefinitely when a combination of phosphoprotein precoated seeds was used together with soluble phosphoproteins in the crystal growth reaction. Second, the phosphoproteins prolonged that stage of the reaction where octacalcium phosphate is the predominant mineral phase present prior to its conversion to the final apatite product. Pre-treatment of the phosphoproteins with calcium diminished their inhibitory activity to seeded crystal growth as well as towards de novo apatite formation in synthetic extracellular fluids. The presence of collagen diminished the inhibitory activity of the phosphoproteins towards de novo precipitation but had no effect on phosphoprotein-modulated apatite crystal growth in the seeded systems. These results suggest a potential regulatory role for phosphoproteins in dentin mineralization.

Animals↗
PubMed1995

Estimation of chronologic age using the aspartic acid racemization method. II. On human cortical bone.

Over the last 20 years a new chemical method, base on the racemization of aspartic acid, has been developed to be used for the estimation of chronologic age in adult individuals. The method has a good accuracy when used on dental enamel, dentine and cartilage. However, in forensic and archeological cases teeth and cartilage are not always available. Since preliminary studies have shown that there are some age-related changes of the D/L aspartic acid ratio also in bone, this study was carried out to further explore if the method could be used for age estimations of bone. Bone samples from 24 individuals, aged 0.2 to 95.6 years were analysed for the D/L ratios with HPLC-technique. Two different fractions of the bone were examined, an acid-soluble peptide fraction and an acid-insoluble collagen-rich fraction. The analyses showed age-related racemizations in both fractions, although of different rates. The correlation coefficients with age were 0.72 in the peptide fraction, and 0.84 in the collagen-rich fraction. It thus seems as if bone may be used for age estimations when more stable tissues like dentine and cartilage are not available.

Adolescent↗
PubMed1992

Microleakage of indirect composite inlays. An in vitro comparison with the direct technique.

Microleakage occurring around directly and indirectly made class-II composite restorations was evaluated on extracted human premolars. Before the evaluation was made, the teeth were thermocycled and stained with an organic dye. Direct class-II composite restorations demonstrated extensive microleakage at their dentin cavosurface margins. Indirect class-II composite inlays luted with a dual-curing composite cement displayed much less microleakage than restorations made using the direct technique. The use of glass-ionomer cement rather than composite cement as the luting material resulted in more microleakage. Rather than being a true microleakage, the leakage of glass-ionomer cement might be the result of a diffusion of the water-soluble organic dye in the water-based cement. The results suggest that the indirect composite technique is superior to the direct technique when it comes to reducing microleakage.

Acid Etching, Dental↗
PubMed1986

Changes in heated and in laser-irradiated human tooth enamel and their probable effects on solubility.

Enamel of intact human teeth laser irradiated in vitro under certain conditions is known to have less subsurface demineralization than unirradiated enamel on exposure to acid; consequently, the potential use of laser irradiance to reduce caries is apparent. The laser-induced physical and/or chemical changes that cause this reduced subsurface demineralization are not known. A laser-irradiated tooth enamel surface will have a temperature gradient that decreases towards the dentin junction. Dependent on irradiant conditions, the temperature may range from greater than 1400 degrees C at the surface to near normal at the dentin-pulp junction. Along this steep temperature gradient, different compositional, structural, and phase changes in the tooth enamel are to be expected. Identification of changes occurring along this gradient has bearing on understanding the dissolution reduction mechanism and, in turn, optimizing its effect. Changes in laser-irradiated material from the highest temperature region have been characterized, but those occurring in sequential layers of decreasing temperatures have not. Since the laser-induced changes are expected to primarily arise from localized heating, previously reported thermally induced changes in tooth enamel on heating in conventional furnaces were utilized to infer corollary changes along the gradient in laser-irradiated tooth enamel. These thermally inferred changes which resulted in modifications in the tooth enamel apatite and/or newly formed phases were correlated with their probable effects on altering solubility. A temperature gradient range from 100-1600 degrees C was considered with subdivisions as follows: I, 100-650 degrees C; II, 650-1100 degrees C; and III, greater than 1100 degrees C. Two of the products formed in range III, alpha-Ca3(PO4)2 and Ca4(PO4)2O, and also identified in the fused-melted material from laser-irradiated tooth enamel, are expected to markedly increase solubility in those regions that contain considerable amounts of these compounds. Products and changes occurring in range II, separate phases of alpha- and/or beta-Ca3(PO4)2 and a modified phase of apatite, may increase or decrease the solubility depending on the Ca/P ratio and the resultant amounts of alpha-, beta-Ca3(PO4)2 formed. Modifications in tooth enamel apatite effected in range I are expected to decrease its solubility; the formation of pyrophosphate in this range may have a substantial effect on reducing the solubility rate.(ABSTRACT TRUNCATED AT 400 WORDS)

Biological Transport, Active↗
PubMed1992

Adhesive properties of modified glass-ionomer cements.

The incorporation of water-soluble polymers and/or vinyl monomers into glass-ionomer cements can yield toughened "hybrid cement-composites". This study compared a commercial water-hardening glass-ionomer cement and seven experimental hybrids in their bonding to both dentin and Silar composite. The cements were sanded and phosphoric-acid-etched or left with an unaltered matrix-formed surface when adhesion to composite was tested. The seven hybrids included: 15% 2-hydroxyethyl methacrylate (HEMA) with appropriate initiators/activators, 29% HEMA, 27% HEMA + 0.5% polyacrylic acid (PAA), 0.5% PAA, 1.5% PAA, 2.5% polyvinyl alcohol, and 2.5% gelatin. Acceptable bond strengths to applied composite and to dentin were observed for most of the modified hybrid cements. There were higher bond strengths with composite when the hybrids were left unetched. Bonding of some unetched, HEMA-containing cements achieved bond strengths (29% HEMA, 10.09 MPa) significantly higher than those of the unmodified cement (4.92 MPa). Resin-modified cements may promote better bonding by improved interaction and compatibility with the resin component of the composite.

Acid Etching, Dental↗
PubMed1999

Osteoblasts/stromal cells stimulate osteoclast activation through expression of osteoclast differentiation factor/RANKL but not macrophage colony-stimulating factor: receptor activator of NF-kappa B ligand.

We previously reported that osteoblasts/stromal cells are essentially involved in the activation as well as differentiation of osteoclasts through a mechanism involving cell-to-cell contact between osteoblasts/stromal cells and osteoclast precursors/osteoclasts. Osteoclast differentiation factor (ODF, also called RANKL/OPGL/TRANCE) and macrophage colony-stimulating factor (M-CSF, also called CSF-1) are two essential factors produced by osteoblasts/stromal cells for osteoclastogenesis. In other words, osteoblasts/stromal cells were not necessary to generate osteoclasts from spleen cells in the presence of both ODF/RANKL and M-CSF. In the present study, we examined the precise roles of ODF/RANKL and M-CSF in the activation of osteoclasts induced by calvarial osteoblasts. Osteoclasts were formed in mouse bone marrow cultures on collagen gel-coated dishes in response to a soluble form of ODF/RANKL (sODF/sRANKL) and M-CSF, and recovered by collagenase digestion. When recovered osteoclasts were further cultured on plastic dishes, most of the osteoclasts spontaneously died within 24 h. Osteoclasts cultured for 24 h on dentine slices could not form resorption pits. Addition of sODF/sRANKL to the recovered osteoclasts markedly enhanced their survival and pit-forming activity. M-CSF similarly stimulated the survival of osteoclasts, but did not induce their pit-forming activity. When primary mouse osteoblasts were added to the recovered osteoclasts, resorption pits were formed on dentine slices. Bone-resorbing factors such as 1alpha,25-dihydroxyvitamin D3, parathyroid hormone, or prostaglandin E2 enhanced pit-forming activity of osteoclasts only in the presence of osteoblasts. M-CSF-deficient osteoblasts prepared from op/op mice similarly enhanced pit-forming activity of osteoclasts. The pit-forming activity of osteoclasts induced by sODF/sRANKL or osteoblasts was completely inhibited by simultaneous addition of osteoprotegerin/osteoclastogenesis inhibitory factor, a decoy receptor of ODF/RANKL. Primary osteoblasts constitutively expressed ODF/RANKL mRNA, and its level was upregulated by treatment with 1alpha,25-dihydroxyvitamin D3, parathyroid hormone, and prostaglandin E2. These results, obtained by using an assay system that unequivocally assesses osteoclast activation, suggest that ODF/RANKL but not M-CSF mediates osteoblast-induced pit-forming activity of osteoclasts, and that bone-resorbing factors stimulate osteoclast activation through upregulation of ODF/RANKL by osteoblasts/stromal cells.

Animals↗
PubMed2006

Effects of resin hydrophilicity on dentin bond strength.

The purpose of this study was to determine if hydrophobic resins can be coaxed into dentin wet with ethanol instead of water. The test hypothesis was that dentin wet with ethanol would produce higher bond strengths for hydrophobic resins than would dentin wet with water. This study examined the microtensile bond strength of 5 experimental adhesives (50 wt% ethanol/50% comonomers) of various degrees of hydrophilicity to acid-etched dentin that was left moist with water, moist with ethanol, or air-dried. Following composite buildups, hourglass-shaped slabs were prepared from the bonded teeth for microtensile testing. For all 3 types of dentin surfaces, higher bond strengths were achieved with increased resin hydrophilicity. The lowest bond strengths were obtained on dried dentin, while the highest bond strengths were achieved when dentin was bonded moist with ethanol. Wet-bonding with ethanol achieved higher bond strengths with hydrophobic resins than were possible with water-saturated matrices.

Analysis of Variance↗
PubMed2005

Intact growth factors are conserved in the extracellular matrix of ancient human bone and teeth: a storehouse for the study of human evolution in health and disease.

For the first time we have extracted, solubilized and identified growth factors, such as insulin growth factor II (IGF-II), bone morphogenetic protein-2 (BMP-2), and transforming growth factor-beta (TGF-beta), from archaeological compact human bone and tooth dentin dating from the late pre-ceramic pottery Neolithic (late PPNB) and the early Middle Ages. These factors are typical of special physiological or pathological situations in the metabolism of bone. The extracellular matrix proteins from bone and teeth of individuals from the late PPNB and early Middle Ages were separated by 2-D electrophoresis and more than 300 different protein spots were detected by silver staining. The matrix protein patterns of compact bone and tooth from the same individual (early Middle Ages) are very different and only 16% of the protein spots were detected in both compact bone and tooth dentin.

Adult↗
PubMed1995

Induction of reparative dentinogenesis in vivo: a synthesis of experimental observations.

EDTA--and/or guanidine HCl--insoluble dentinal matrix, or demineralized dentin which had been treated with plasma fibronectin, or pieces of Millipore filters coated with a recombinant fibronectin-like engineered polymer, incorporating many RGD sequences, were implanted into central parenchymal sites of young dog molars, via mechanical pulp exposures. Furthermore demineralized dentin and Millipore filters coated with plasma fibronectin were placed into the central pulp of old animals. Histological analysis of buffered formalin-fixed tissues showed that: 1. The dentinogenic activity was retained in the EDTA--and/or guanidine-insoluble dentin matrix. 2. Implantation of Millípore filters supplemented with the recombinant polymer did not induce any odontoblast-like cell differentiation, indicating that the interactions of pulp cells with the exogenous fibronectin are not RGD-dependent. 3. Acid-insoluble dentin matrix or plasma fibronectin (both separately inducing dentinogenesis in dental pulp of young animals) did not show any dentinogenic activity when exposed in pulp sites of old animals. Acid-insoluble dentin matrix and plasma fibronectin also failed to induce dentinogenic activity in the young pulpal tissues, when both factors were combined before to their implantation. Synthesizing the present data with previous relevant information it could be suggested that in the mechanism initiating reparative dentinogenesis, growth factors (endogenous or artificially implanted) and fibronectin are involved and this mechanism seems to be more complex than the simple immobilization of pulp cells onto an adhesion substratum.

Aging↗
PubMed2003

Acidic solubility of luting cements.

OBJECTIVES: Chemical disintegration of luting cements can adversely affect their long term success. The aim of the present study was to assess the susceptibility of zinc phosphate cement, glass ionomer cement and resin cement to erosion at various pH values. METHODS: Zinc phosphate and glass ionomer cement samples were eroded in 0.3% citric acid adjusted to pH 3.0, 4.0, 5.0, 6.0 and 7.0 at a temperature of 35 degrees C. A control group was stored in saline. Profilometric readings were performed after 30 min and 1h erosion time. Resin cement samples were eroded at pH 3.0 with measurements after 30 min, 6h, 24h and 7 days. A saline control group was measured after 7 days. RESULTS: Erosion of zinc phosphate cement was significantly higher at all pH values than erosion of glass ionomer cement with erosion depth increasing with decreasing citric acid pH. No erosion was seen in the control groups. The resin cement did not show any change in surface profile after the 7 day erosion period compared to the control group. Comparison to dental tissues revealed that erosion of glass ionomer cement in vitro was slightly lower than erosion of enamel and dentine. In contrast, erosion of zinc phosphate cement was much higher.

Acids↗
PubMed1998

Physical properties of resin-modified glass-ionomers.

The physical properties and shear bond strength of two resin-modified glass-ionomers: RMGIs (FJL and VT) and as a control, a conventional glass-ionomer: CGI (FJ), were evaluated. Ninety percent of the setting shrinkage under dry conditions and nominal hygroscopic expansion in water during 360 h occurred within the first 24 h. The nominal hygroscopic expansion of CGI was 0.38% while the RMGIs demonstrated over 1%, but considering the setting shrinkage the total hygroscopic expansion of CGI and the RMGIs were over 7 times and about 2.5 times, respectively. The thermal expansion of the RMGIs after water immersion was greater than that under dry condition. In contrast, FJ demonstrated expansion less than under dry condition. There was no significant difference between FJL and VT (P < 0.05) as regard water absorption. The shear bond strength of the RMGIs to enamel and dentin were superior to FJ, except for VT to dentin.

Absorption↗
PubMed1989

[Bonding of MMA-BPO. DMPT resin to bovine tooth coated by poly (methyl methacrylate-co-p-styrene sulfonic acid)].

The objective of this study was to prepare a new type water-soluble bonding agent, methyl methacrylate (MMA)-p-styrene sulfonic acid copolymer (MS), and to investigate the effect of MS on bonding between resins and tooth substrates. MS is cross-linked with Ca2+ released from ground enamel and dentin and could be immobilized on their surface. A sample was prepared by bonding an acrylic rod with a BPO-amine catalyzed self-curing resin to ground enamel and dentin coated with an aqueous mixture of FeCl3 and 10 wt% MS. After immersion in water for 24 hrs, the tensile bond strength was measured. The bond strength to both enamel and dentin was only 2 MPa and adhesive failure occurred at the interface between cured MS and self-curing resin. This suggested that cured MS could adversely effect the polymerization of self-curing resins. A second treatment of cured MS on the tooth surface with metallic cations was carried out to minimize the amount of free sulfonic acids in the MS disturbing radical formation in self-curing resin. The second treatment improved the bond strength to 6 MPa.

Acrylic Resins↗
PubMed2004

Identification of collagen encapsulation at the dentin/adhesive interface.

PURPOSE: The purpose of this study was to investigate the dentin/adhesive interfacial characteristics of three current commercial adhesives with different relative hydrophilic/hydrophobic composition, using a nondestructive staining technique. MATERIALS AND METHODS: Dentin surfaces of 18 unerupted human third molars were randomly selected for treatment with one of three commercial dentin bonding agents according to manufacturers' instructions for the "wet" bonding technique. The adhesives were ranked based on hydrophilic/hydrophobic component ratios (ie, ability to dissolve in water), highest to lowest, as follows: Uno (Pulpdent) > Prime&Bond NT (PBNT, Dentsply Caulk) > Single Bond (SB, 3M ESPE). Dentin/adhesive (d/a) interface sections were stained with Goldner's trichrome, a classical bone stain, and examined using light microscopy. RESULTS: The extent and degree to which the adhesive encapsulates the demineralized dentin matrix is reflected in the color differences in the stained sections. The depth of demineralization appeared comparable among these bonding systems, but adhesive infiltration varied from highest to lowest as follows: Uno > PBNT > SB. CONCLUSIONS: The differential staining technique provided a clear representation of the depth of dentin demineralization and extent/degree of adhesive encapsulation of the exposed collagen at the d/a interface. This technique provides a mechanism for readily identifying vulnerable sites at the d/a interface. The composition of the one-bottle adhesive systems has a substantial effect on the interfacial structure of the d/a bond.

Acid Etching, Dental↗
PubMed1993

The effects of adriamycin on dental proteins formation and mineralization in vitro.

Second maxillary molars of 4-5 days old golden hamsters were exposed for 2 h in vitro to 1 mg/L adriamycin, rinsed and subsequently cultured up to 7 days without the drug. At days 3, 5 or 7 of culture the synthesis of extracellular tooth matrices and their mineralization were examined by measuring the incorporation of 3H-proline and the uptake of 45Ca and 32PO4 by the explants during a 24 h pulse labeling. Compared with unexposed control explants, exposure to adriamycin for the first 2 h of culture had no effect on total biosynthesis of proline-containing matrix proteins. However, at days 3 and 5 of culture it increased the quantity of water-soluble enamel matrix proteins (amelogenins). Adriamycin also strongly reduced the amount of organically-bound 32P-activity in a fraction extractable with guanidine-HC1-EDTA only, allegedly containing a mixture of mineral-associated proteins from both enamel and dentin. Since this decrease of 32P-activity coincided with the formation of osteodentin in the pulp as shown previously in histological and electron microscopical studies, it was speculated that osteodentin matrix may not contain the highly phosphorylated, dentin-specific phosphoproteins (DPP). Adriamycin also affected the uptake of 45Ca and 32PO4. At day 3 these values were slightly higher than control values but lower at days 5 and 7. It therefore appears that a 2 h exposure to adriamycin in concentrations as low as 1 mg/L causes an acceleration of secretory amelogenesis by tooth germs in vitro. It also induces pulp cells to form osteodentin.

Animals↗
PubMed2006

Role of human pulp fibroblasts in angiogenesis.

After pulp amputation, complete pulp healing requires not only reparative dentin production but also fibroblast proliferation, nerve fiber growth, and neoangiogenesis. This study was designed to investigate the role of pulp fibroblasts in angiogenesis. Human pulp fibroblasts from third molars co-cultured with human umbilical vein endothelial cells induced the organization of endothelial cells and the formation of tubular structures corresponding to capillaries in vivo. The direct contact between both cells was not necessary to induce angiogenesis, and the observed effect was due to soluble factors. This was confirmed with neutralizing antibodies against FGF-2 and VEGF, which decreased the angiogenic effects of these soluble factors. Immunohistochemistry showed that both FGF-2 and VEGF were expressed in human dental pulp fibroblasts, and this expression increased after injury. These results suggest that the pulp fibroblasts secrete angiogenic factors, which are necessary for complete pulp healing, particularly at the pulp injury site.

Adolescent↗
PubMed1999

Predicting protein decomposition: the case of aspartic-acid racemization kinetics.

The increase in proportion of the non-biological (D-) isomer of aspartic acid (Asp) relative to the L-isomer has been widely used in archaeology and geochemistry as a tool for dating. the method has proved controversial, particularly when used for bones. The non-linear kinetics of Asp racemization have prompted a number of suggestions as to the underlying mechanism(s) and have led to the use of mathematical transformations which linearize the increase in D-Asp with respect to time. Using one example, a suggestion that the initial rapid phase of Asp racemization is due to a contribution from asparagine (Asn), we demonstrate how a simple model of the degradation and racemization of Asn can be used to predict the observed kinetics. A more complex model of peptide bound Asx (Asn + Asp) racemization, which occurs via the formation of a cyclic succinimide (Asu), can be used to correctly predict Asx racemization kinetics in proteins at high temperatures (95-140 degrees C). The model fails to predict racemization kinetics in dentine collagen at 37 degrees C. The reason for this is that Asu formation is highly conformation dependent and is predicted to occur extremely slowly in triple helical collagen. As conformation strongly influences the rate of Asu formation and hence Asx racemization, the use of extrapolation from high temperatures to estimate racemization kinetics of Asx in proteins below their denaturation temperature is called into question. In the case of archaeological bone, we argue that the D:L ratio of Asx reflects the proportion of non-helical to helical collagen, overlain by the effects of leaching of more soluble (and conformationally unconstrained) peptides. Thus, racemization kinetics in bone are potentially unpredictable, and the proposed use of Asx racemization to estimate the extent of DNA depurination in archaeological bones is challenged.

Aspartic Acid↗
PubMed2006

The effect of calcium hydroxide on solubilisation of bio-active dentine matrix components.

Calcium hydroxide (Ca(OH)(2)) has been used extensively to induce dentine regeneration through formation of dentine bridges at sites of pulp exposure after dental tissue injury, however, the biological processes underpinning these events are unclear. We hypothesise that growth factors and other bio-active molecules, sequestered within dentine matrix, may be released by the action of Ca(OH)(2) and signal gene expression in pulp cells, which mediates the changes in cell behaviour observed during regeneration. Powdered sound, human dentine samples were extracted with either 0.02 m Ca(OH)(2), pH 11.7 or 10% EDTA, pH 7.2 ( a control known extractant of bio-active and other ECM molecules from dentine) over a 14-day period. Extracts were compared for non-collagenous protein (NCP) and glycosaminoglycan (GAG) content using dye binding assays and protein compositions were analysed by 1D-polyacrylamide gel electrophoresis (1D-PAGE) and TGF-beta1 ELISA. The effects of extracts on TGF-beta1, Collagen-1alpha and Nestin gene expression were analysed using semi-quantitative RT-PCR in the dental MDPC-23, OD-21 and fibroblastic Swiss 3T3 cell lines following 24h of exposure. Ca(OH)(2) solubilised NCPs and GAGs from the dentine ECM, although with a lower yield than the EDTA solution and with different kinetics. 1D-PAGE analysis demonstrated some differences in profiles for proteins solubilised from dentine by Ca(OH)(2) and EDTA. Both solutions released TGF-beta1 from the dentine with higher concentrations present in the EDTA (1.395 +/- 0.036 ng/mg) versus the Ca(OH)(2) (0.364 +/- 0.012 ng/mg) extract. Notably, both extracts induced similar gene expression profiles in all cell lines. These data provide a rational explanation for the action of Ca(OH)(2) during pulp capping in which the cellular activities involved in dentine bridge formation may be mediated through release of growth factors and other bio-active molecules from the dentine by Ca(OH)(2).

Base Sequence↗
PubMed1985

Short-term effects of fluoride on biosynthesis of enamel-matrix proteins and dentine collagens and on mineralization during hamster tooth-germ development in organ culture.

The effect of various concentrations of fluoride (F-) on cell proliferation, matrix formation and mineralization was examined in hamster molar tooth germs in premineralizing and mineralizing stages. The exposure lasted 16 h (mineralizing stages) and 24 h (premineralizing stages) and the F- levels ranged from 2.63 microM to 2.63 mM; [3H]-thymidine, [3H]-proline, 45Ca and 32PO4 were used as markers for cell proliferation, matrix formation and mineralization, respectively. The proline-labelled amelogenins were isolated by sequential extraction with water and formic acid and their nature examined by SDS-urea-polyacrylamide electrophoresis. Digestion by collagenase was used to assess the amount of proline incorporated into collagens. F- in concentrations up to 1.31 mM inhibited neither biosynthesis of DNA and amelogenins, nor synthesis of collagens and their hydroxylation. Amelogenins extracted from F- induced, non-mineralizing enamel matrix had the same electrophoretic mobility and the same degree of phosphorylation as amelogenins from normal, mineralizing enamel. However, F- increased the uptake of 45Ca and TCA-soluble 32P dose-dependently, starting with 52 microM. Thus, interference with secretion of enamel matrix by F- takes place at much lower concentrations than required to inhibit biosynthesis of enamel matrix.

Animals↗