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Bovine dentin phosphophoryn: composition and molecular weight.

The molecular weight of phosphophoryn, an acidic phosphoprotein unique to dentin matrix, has been difficult to determine because of a combination of neutral protease activities in this tissue and the intrinsic high charge density of the molecule. In this study, bovine dentin phosphophoryn (BDPP) was isolated by a procedure designed to prevent proteolysis. Bovine unerupted third molar powder was demineralized by ethylenediaminetetraacetic acid (EDTA). The EDTA-soluble phosphophoryn fraction was isolated and purified by sequential calcium chloride precipitation, gel filtration in sodium dodecyl sulfate (NaDodSO4) containing buffer, anion-exchange chromatography, and finally gel filtration in 4 M guanidine hydrochloride (4 M Gdn.HCl) buffer. Sedimentation equilibrium, sedimentation velocity, and diffusion coefficient data, viscosity studies in a high ionic strength buffer, and NaDodSO4 gradient gel electrophoresis data gave consistent results for the molecular weight of BDPP, all being in the range of 151 000-167 000. This range is much higher than any previously reported value. An anomalous behavior was observed in nongradient NaDodSO4 gel electrophoresis. Dissociative analytical gel filtration chromatography in 4 M Gdn.HCl gave a molecular weight value of 100 000. This discrepancy was resolved by studying the viscosity of BDPP in 4 M Gdn.HCl which showed BDPP does not assume a true random-chain conformation in this solvent.

Amino Acids↗

Collagen mediates adhesion of Streptococcus mutans to human dentin.

Some strains of Streptococcus mutans were found to recognize and bind collagen type I. Binding of 125I-labeled collagen type I was specific in that collagen types I and II, but not unrelated proteins, were able to inhibit binding of the labeled ligand to bacteria. Collagen binding to S. mutans was partially reversible and involved a limited number of bacterial binding sites per cell. S. mutans UA 140 cells bound collagen type I with high affinity (Kd = 8 x 10(-8) M). The number of binding sites per cell was 4 x 10(4). Collagen-binding strains of S. mutans were found to adhere to collagen-coated surfaces as well as to pulverized root tissue. S. mutans strains that did not bind the soluble ligand were unable to adhere to these substrata. Adherence to collagen-coated surfaces could be inhibited with collagen or clostridial collagenase-derived collagen peptides. Adherence of S. mutans to dentin was enhanced by collagen types I and II but inhibited by collagen peptides. S. mutans UA 140 bound significantly less 125I-collagen type I following treatment with peptidoglycan-degrading enzymes. These enzymes released a collagen-binding protein (collagen receptor) with a relative molecular size of 16 kDa. The results of this study suggest that collagen mediates adhesion of S. mutans to dentin. This interaction may target collagen-binding strains of S. mutans to dentin in the oral cavity and may play a role in the pathogenesis of root surface caries.

Adhesins, Bacterial↗

The influence of some dentin primers on calcium hydroxide lining cement.

UNLABELLED: Dentin primer is applied as a routine procedure prior to bonding to improve the sealing properties of direct polymerizing resins. Some primers contain acetone or alcohol that may affect the properties of calcium hydroxide liner which is placed as a direct or indirect pulp cap. If calcium hydroxide is softened or smeared over the cavity walls, the bonding will be impaired. Therefore, if this occurs, the cement must be removed, the walls must be cleansed, and the procedure must be repeated with careful application of dentin primer. OBJECTIVE: The purpose of this study was to determine the wear and compressive strength of a calcium hydroxide liner after exposure to different kinds of dentin primers for different periods of time. METHODS: The calcium hydroxide used in this study was Dycal. It was mixed according to the manufacturer's instructions and placed in plastic rings of 0.5 mm x 5 mm and allowed to set at 37 degrees C for 15 min under 500 gm load. To determine erosion, the height for each sample before and after application of primers was recorded using a Digital Height Measuring Instrument "Digmar" 817. Compressive strength specimens were also prepared. RESULTS: Calcium hydroxide treated with Optibond (alcohol based) or Syntac (acetone based) for 1 min or 5 min had the highest erosion values and the lowest compressive strength values. Gluma CPs (water based primer) had the least effect on calcium hydroxide values.

Analysis of Variance↗

Effects of thermocycling on the tensile bond strength between resin cement and dentin surfaces after temporary cement application.

PURPOSE: In a previous study the authors found that 2 of 3 tooth conditioners examined were able to reduce the negative effect of temporary cement on the bond strength between resin cement and teeth. The aim of this study was to evaluate the bond durability with the conditioners, as well as their capability for temporary cement dissolution. MATERIALS AND METHODS: After eliminating the temporary cement with a curette from the bovine dentin surface, a conditioner (ethyl dihydrogen phosphate, EP; or methacryloxyethyl dihydrogen phosphate, MEP) was applied to the surface and a resin cement was adhered. Tensile bond strength measurement, scanning electron microscopic observation, and energy-dispersive analysis were carried out without thermocycling and after thermocycling. RESULTS: The tensile bond strength gradually decreased with an increase in the number of thermocycles. However, specimens treated with EP or MEP showed significantly higher tensile bond strength values compared to those that did not receive conditioner application (P < 0.0001); EP showed significantly higher tensile bond strength values compared to the specimens that did not receive both temporary cement and conditioner application (P < 0.0001). The scanning electron microscopic, energy-dispersive, and solubility analyses suggest that both conditioners have the capability of dissolving temporary cement remnants. CONCLUSION: Both EP and MEP have potential value as conditioners to reduce the negative effect of temporary cement on the bond strength between resin cement and dentin and to improve the bond durability.

Analysis of Variance↗

MC3T3-E1-conditioned medium-induced mineralization by clonal rat dental pulp cells.

Dental pulp is thought to participate in supplementary mineralization, such as reparative dentin and pulp stones, but no direct proof of this has been reported. To study this process at a molecular level, we investigated the matrix mineralization of dental pulp using a clonal cell line (RPC-C2A) derived from rat incisor dental pulp. Mineralized nodules in extracellular matrix were formed by RPC-C2A cells cultured in the presence of conditioned medium (CM) from confluent osteoblastic MC3T3-E1 cells. These nodules were stained by the von Kossa method and with alizarin red S and quantified by the measurement of acid-soluble calcium deposition. This CM was most effective when collected 3-6 days after confluency and added at 50% to the culture medium. The CM-treated RPC-C2A cells showed high alkaline phosphatase activity, a high mRNA level of osteocalcin and decreases in the mRNA levels of osteopontin and osteonectin, but undetectable levels of mRNA of dentin sialophosphoprotein by Northern blot analyses. A pan-specific anti-transforming growth factor (TGF)-beta antibody and a soluble form of receptor for bone morphogenetic protein (BMP)-2/-4 did not neutralize the CM-induced mineralization. These results suggest that some soluble factor(s) other than TGF-beta or BMP-2/-4 in the CM from MC3T3-E1 cells cause differentiation of RPC-C2A cells to osteoblast-like cells.

Alkaline Phosphatase↗

In vivo and in vitro study of 90Sr in developing rat molar enamel.

The uptake patterns of 90Sr in developing rat molar enamel were studied in vivo and in vitro. Autoradiographic methods were used that preclude loss or translocation of tracers associated with water-soluble compounds in the sections. In eight-day-old rats injected with the tracer, 90Sr uptake in the enamel was significantly less than for dentin and bone, particularly at early sacrifice times. The uptake pattern of 90Sr was somewhat different from that previously observed for 45Ca. The in vitro experiments indicated that the viable intact enamel organ limits uptake of 90Sr by enamel in both the secretory and maturation phases of enamel formation.

Amelogenesis↗

Joint fluid from patients with failed total hip arthroplasty stimulates pit formation by mouse osteoclasts on dentin slices.

Periprosthetic bone resorption has been implicated in the failure of total joint arthroplasty. Osteolysis is reported to be associated with bone resorption induced by bone-resorbing cytokines, which are released from macrophages and fibroblasts in periprosthetic tissues after stimulation by wear debris generated in the joint cavity. Recent reports have suggested the concept of the effective joint space, which includes all periprosthetic regions that are accessible to joint fluid and wear debris. In this study, we examined the levels of interleukin-6 (IL-6), soluble IL-6 receptor (sIL-6R), and tartrate-resistant acid phosphatase (TRAP) in joint fluid after failed total hip arthroplasty (THA) with osteolysis and investigated whether the joint fluid could activate osteoclastic bone resorption using unfractionated mouse bone cells cultured on dentin slices. Histochemical analysis showed the presence of more TRAP-positive cells in synovial capsules from failed THA patients when compared with osteoarthritis (OA) patients (controls). The levels of IL-6, sIL-6R, and TRAP in joint fluid from failed THA patients were significantly higher than in OA patients. Mouse osteoclasts cultured on dentin slices with joint fluid from failed THA patients with osteolysis produced a significant increase of pit area, whereas cells cultured with joint fluid from OA patients did not. Interestingly, osteoclastic bone resorption on dentin slices was significantly correlated with TRAP activity in joint fluid (p < 0.0001). These results suggest that joint fluid containing bone-resorbing cytokines is produced by synovial capsules in failed THA patients with osteolysis and may activate osteoclasts around the prosthesis in combination with those produced by interface tissues, thus contributing to periprosthetic bone resorption.

Acid Phosphatase↗

Activation of osteoclast-mediated bone resorption by the supernatant from a rabbit synovial cell line in response to polyethylene particles.

It is unknown whether the soluble factors produced from cells activated by wear particles in the fibrous tissue around failed joint prostheses really activate osteoclastic bone resorption. In this study, the activation of osteoclast-mediated bone resorption by the products from a rabbit synovial cell line (HIG-82) stimulated by various particles was investigated using rabbit unfractionated bone cells cultured on a dentin slice. The HIG cells were challenged with the following laboratory-made particles: high-density polyethylene (PE), cobalt alloy (Co-Cr), titanium alloy (Ti6Al4V), pure titanium (Ti), and sintered hydroxyapatite (HA). The size of each particle was < 2 microns. The supernatants from HIG cells cultured with the appropriate concentration of wear particles were added to unfractionated bone cells on a dentin slice, and then resorbed areas were determined for each particle. Interestingly, resorbed areas significantly increased only when the culture medium from HIG cells with PE particles was added to unfractionated bone cells. This study demonstrates that PE particles stimulate the rabbit synovial cells to produce soluble factors that induce osteoclast-mediated bone resorption. Moreover, this experimental model is a useful method sensitively to evaluate the effects of soluble factors from the cells stimulated by particulate biomaterials from joint prostheses on osteoclastic bone resorption.

Animals↗

Physical and chemical properties of a new root-end filling material.

This study determined the chemical composition, pH, and radiopacity of mineral trioxide aggregate (MTA), and also compared the setting time, compressive strength, and solubility of this material with those of amalgam, Super-EBA, and Intermediate Restorative Material (IRM). X-ray energy dispersive spectrometer in conjunction with the scanning electron microscope were used to determine the composition of MTA, and the pH value of MTA was assessed with a pH meter using a temperature-compensated electrode. The radiopacity of MTA was determined according to the method described by the International Organization for Standardization. The setting time and compressive strength of these materials were determined according to methods recommended by the British Standards Institution. The degree of solubility of the materials was assessed according to modified American Dental Association specifications. The results showed that the main molecules present in MTA are calcium and phosphorous ions. In addition, MTA has a pH of 10.2 initially, which rises to 12.5 three hours after mixing. MTA is more radiopaque than Super-EBA and IRM. Amalgam had the shortest setting time (4 min) and MTA the longest (2 h 45 min). At 24 h MTA had the lowest compressive strength (40 MPa) among the materials, but it increased after 21 days to 67 MPa. Finally, except for IRM, none of the materials tested showed any solubility under the conditions of this study.

Aluminum Compounds↗

Endodontic irrigation.

This review of current research and literature investigates trends in endodontic irrigation, focusing on the goals of irrigation and the ability of different irrigants to meet these goals.

Anti-Infective Agents, Local↗

Effect of resin hydrophilicity on water-vapour permeability of dental adhesive films.

This study examined the water-vapour permeability of thin polymerized resin films fabricated from five co-monomer blends of increasing degrees of hydrophilicity, as measured by their Hoy's solubility parameters. Neat resin films were prepared from five experimental light-curable resins (n = 10). Each film was mounted in a Fisher permeability cup with 8 g of water placed inside the cup. The experiments were conducted in a modified twin-outlet desiccator connected to a vacuum pump in one outlet to permit a continuous airflow to encourage water evaporation. Weight losses by water evaporation were measured at 3, 6, 9, 24, 30, and 48 h by using an analytical balance. Additional resin films were examined by using transmission electron microscopy (TEM) after immersion in ammoniacal silver nitrate. A significant correlation was observed between the cumulative water loss at 48 h and the Hoy's total cohesive energy density (delta(t)). Transmission electron microscopy revealed silver-filled channels along film peripheries and silver grains of decreasing dimensions toward the film centres in co-monomer blends 3, 4, and 5 of increasing hydrophilicity. Hydrophilic dentin adhesives polymerized in thin films are prone to water loss by evaporation. This probably accounts for the water droplets seen on the surface of vital-bonded dentin after the application of simplified dentin adhesives.

Dentin-Bonding Agents↗

Substance P stimulates release of RANKL via COX-2 expression in human dental pulp cells.

OBJECTIVE: Our previous study found that substance P (SP), a sensory neuropeptide, was expressed in the dental pulp of rats during experimental tooth movement. We examined the effects of SP on the production of prostaglandin (PG) E2 and the receptor activator of nuclear factor- B ligand (RANKL) by human dental pulp fi broblast-like (HDPF) cells. MATERIALS AND METHODS: SP was added to cultured HDPF cells at concentrations ranging from of 10(-4) to 10(-12) mol/L. PGE2 and soluble RANKL (sRANKL) levels were determined using enzyme-linked immunosorbent assay kits. Gene expression was confi rmed by RT-PCR analysis. Pit formation assays using dentin slices were carried out to examine the effect of SP on osteoclastogenesis. RESULTS: The levels of PGE2 and sRANKL increased in the presence of SP, though the increases were greater in the experimental groups in both a time- and concentration-dependent manner, and the increase of RANKL was partially mediated by PGE2. The gene expression of cyclooxygenase (COX)-2 and RANKL was up-regulated, and conditioned medium samples obtained from HDPF cells treated with SP induced bone resorption. CONCLUSIONS: SP stimulated the production of PGE2 and RANKL, and promoted bone resorption. Therefore, SP may be involved in pulpal inflammation and root resorption during orthodontic tooth movement.

Adolescent↗

Understanding the chemistry of dental erosion.

The mineral in our teeth is composed of a calcium-deficient carbonated hydroxyapatite (Ca10-xNax(PO4)6-y(CO3)z(OH)2-uFu). These substitutions in the mineral crystal lattice, especially carbonate, renders tooth mineral more acid soluble than hydroxyapatite. During erosion by acid and/or chelators, these agents interact with the surface of the mineral crystals, but only after they diffuse through the plaque, the pellicle, and the protein/lipid coating of the individual crystals themselves. The effect of direct attack by the hydrogen ion is to combine with the carbonate and/or phosphate releasing all of the ions from that region of the crystal surface leading to direct surface etching. Acids such as citric acid have a more complex interaction. In water they exist as a mixture of hydrogen ions, acid anions (e.g. citrate) and undissociated acid molecules, with the amounts of each determined by the acid dissociation constant (pKa) and the pH of the solution. Above the effect of the hydrogen ion, the citrate ion can complex with calcium also removing it from the crystal surface and/or from saliva. Values of the strength of acid (pKa) and for the anion-calcium interaction and the mechanisms of interaction with the tooth mineral on the surface and underneath are described in detail.

Calcium↗