Chemical interaction of 4-META with enamel in resin-enamel bonds.

X-ray photoelectron spectroscopy (XPS) is used to analyze 4-META resin and enamel that are debonded at an adhesive interface. The XPS spectra showed two chemical states for Ca: one resulted from Ca of hydroxyapatite and the other, an unknown chemical state, suggested that Ca was chemically bonded with 4-META. We postulate that for a chemical reaction of 4-META and hydroxyapatite, the chemical structure of carboxyl groups will resemble that of calcium phthalate. Hence, calcium phthalate was used as a reference material. Additionally, the spectra obtained from the adhesive interface and the mixture of calcium phthalate with hydroxyapatite particles were compared using peak deconvolution analysis. XPS analysis revealed that the chemical bond of 4-META with enamel resembled the chemical state of Ca in calcium phthalate. Consequently, we suggest that Ca of the enamel and the carboxyl group of 4-META were chelate-bonded at the interface.

Viscoelastic adlayers of collagen and lysozyme studied using quartz crystal microbalance with dissipation monitoring.

In this study the effects of molecular structure of proteins on their adsorption behaviour and viscoelasticity are investigated using a QCM-D method. The adsorption measurements show that spherical lysozyme is rapidly adsorbed on the gold surface as a stiff monolayer, as indicated by a sharp drop in the oscillation frequency (Deltaf) of the sensor, and by a very small increase in the energy dissipation of the adlayer (DeltaD). Fibrous calfskin collagen (CSC) is, however, adsorbed on the same surface rather slowly in two steps to form a thick and soft multilayer (large Deltaf and DeltaD) at pH 3 in salt-free conditions. The two-step adsorption is not observed for stiffly aggregated CSC. This study clearly demonstrates that the polymer structure strongly affects not only how adsorption develops but also the viscoelastic properties of the adlayer.

Microscopic observations and inflammatory cytokine productions of human macrophage phagocytising submicron titanium particles.

This study was performed to microscopically observe and measure inflammatory cytokine production by human macrophages phagocytosing submicron titanium (Ti) particles. Observations with secondary electron microscopy (SEM), SEM/electron probe microanalysis (EPMA) and transmission electron microscopy (TEM) indicated that macrophages [phorbol-12-myristate-13-acetate (PMA)-differentiated THP-1 cells] at 24 h in culture actively phagocytosed and accumulated submicron Ti particles in intracellular phagosomes, in which refinement of Ti particles occurred. The macrophages were also cultured for 24 h in four media with and without submicron Ti particles and lipopolysaccharide (LPS; components of bacteria). Whilst neither stimulus reduced cell viability, submicron Ti particles and LPS activation independently and synergistically caused the macrophages to produce three inflammatory cytokines (TNF-alpha, IL-1beta and IL-6) at high levels in the culture supernatants. The inflammatory and osteolysis conditions caused by macrophages phagocytosing submicron Ti particles would be worsened by challenge with LPS in patients wearing Ti prostheses.

Gene expression analyses of human macrophage phagocytizing sub-micro titanium particles by allergy DNA chip (Genopal).

The purpose of this study was to examine gene expressions of macrophage phagocytizing sub-micro Ti particles by a DNA chip. Human monocytic cell line THP-1 was differentiated into macrophages by culturing for two days in medium supplemented with 200 nM phorbol ester (PMA). The macrophages were then cultured in four media: medium without PMA (control); medium with suspended sub-micro Ti particles (0.5 wt%); medium with 1.0 microg/ml lipopolysaccharide (LPS); and medium with LPS and Ti particles. After 6 hours' culture, total RNA were extracted and gene expressions were evaluated by DNA allergy chip with 205 allergy and inflammation related gene spots. We found that phagocytosis of sub-micro Ti particles and LPS independently and synergistically up-regulated 17 inflammation-related genes more than two-fold. The extensive expressions of four genes (CCL1, IL1B, IL6 and IL8) were further confirmed by real-time quantitative PCR. It turned out that dual stimulation of LPS and Ti particles most up-regulated three genes (IL1B, IL6 and IL8), followed by LPS while Ti particles moderately but least increased, suggesting that phagocytosis of sub-micro Ti particles induces moderate inflammation with its degree less than LPS, but phagocytosis of sub-micro Ti particles has the potential to worsen inflammation caused by LPS-stimulated macrophages.

Preparation and in vivo evaluation of apatite/collagen packed composite by alternate immersion method and Newton press.

Further development of bio-compatible, bio-absorbable, and osteo-conductive bio-materials is desired for bone grafts in dental and medical clinics. One candidate material might be a high-density apatite/collagen composite, which cures relatively large bone defects. To produce such a composite, we freeze-dried type I collagen solution, cross-linked the formed sponge by 2 wt % glutaraldehyde, immersed the insoluble sponge in CaCl(2) and Na(2)HPO(4) solutions alternately five times, and compacted the sponge by Newton press at 5000 kgf. For comparison, cross-linked collagen without alternate immersion was also pressed. SEM/EPMA, XRD, and FTIR analyses clarified that alternate immersion successfully coated the collagen sponge with hydroxyapatite. Packed apatite/collagen composite and collagen disks 6 mm in diameter and 0.5 mm in height were implanted in the subperiostea of rabbit tibiae for 2, 4, 8, and 12 weeks to assess bio-compatibility, bio-absorbability, and osteo-conductivity. Histological observations showed that the packed apatite/collagen composite was biocompatible, osteo-conductive for up to 8 weeks, and largely bio-absorbed at 12 weeks, while the packed collagen sponge caused an undesirable foreign body reaction, which worsened with the implantation period. The overall findings suggest that this packed apatite/collagen composite might be used as a new bio-absorbable bone graft material.

Effect of pH and addition of salt on the adsorption behavior of lysozyme on gold, silica, and titania surfaces observed by quartz crystal microbalance with dissipation monitoring.

The adsorption behaviors of lysozyme on dentally related Au, SiO2, and TiO2 surfaces were investigated by a quartz crystal microbalance with dissipation monitoring (QCM-D) method. Frequency shifts indicated that while lysozyme (pI 11) was fairly adsorbed on the SiO2 (pI 1.9) surface at both pH 3 and 7, it was adsorbed on TiO2 (pI 6.3) surface only at pH 7. However, adsorption was disturbed by 50 mM NaCl. These results strongly suggested an electrostatic nature of the adsorption behavior. Though a large-scale adsorption of the lysozyme on Au sensor was pH-insensitive, softness of the adlayer as seen from the dissipation profile was pH-dependent, indicating an interaction of another type. With all the surfaces, the small dissipation change indicated a stiff lysozyme adlayer. Results of this study revealed that the controlled electrostatic interaction between the material surface and lysozyme might be a useful method for imparting antibacterial property to the dental materials.

Dose-dependent effects of Ni (II) ions on production of three inflammatory cytokines (TNF-alpha, IL-1beta and IL-6), superoxide dismutase (SOD) and free radical NO by murine macrophage-like RAW264 cells with or without LPS-stimulation.

The effect of Ni (II) ions on macrophages is not well understood. The purpose of this study was to examine the dose-dependent effects of Ni (II) ions up to 1,000 micromol/L on production of three inflammatory cytokines (TNF-alpha, IL-1beta and IL-6), superoxide dismutase (SOD) and nitric oxide (NO) by murine macrophage-like RAW264 cells with (+) or without (-) lipopolysaccharide (LPS) -stimulation. Ni (II) ions caused LPS (-) RAW264 cells to slightly increase production of TNF-alpha and IL-6, proportionally to the Ni (II) ion concentration while IL-1beta was not produced, and to slightly increase production of SOD and NO. It can be concluded that Ni (II) ions dose-dependently increased the inflammatory and oxidative stress conditions of LPS (-) RAW264 cells. LPS-stimulation caused RAW264 cells to produce in abundance the three inflammatory cytokines, SOD and NO. Ni (II) ions dose-dependently reduced the three cytokine quantities and NO amounts in LPS (+) RAW264 cells, while dose-independently increasing SOD amounts. It was noted that Ni (II) ions dose-dependently reduce the resistance power against bacteria of LPS (+) macrophages, because the production of volatile NO--bacteria killer is diminished proportionally to the Ni (II) ion concentration.

New index for the stability of a type I collagen affected by hydrophobic environment.

Effects of hydrophobic environment adjusted by various alcohols on the structural stability of calfskin collagen (CSC) were studied to elucidate the nature of collagen-monomer interaction in adhesion. The stability of CSC in aqueous alcohol solutions was represented by its denaturation temperature, Td, measured by DSC. The hydrophobicity of the alcohol solutions was quantified with their specific dielectric constants, epsilon(r), calculated from their concentrations. The effect of each alcohol to stabilize or destabilize CSC was evaluated by the initial slope of each Td vs. epsilon(r) plot, denoted as -(dTd/d epsilon(r))ini and termed as stabilization power. Results showed that a hydrophobic environment with a smaller epsilon(r) lowered the stabilization power. Stabilization power ranged from -3 (strong destabilization) for phenol (epsilon(r) =12) to +0.3 (weak stabilization) for glycerol (epsilon(r)=47). In view of the encouraging results obtained in this study, the new index was therefore helpful in predicting the effects of new dental materials of known epsilon(r) values on the stability of dentinal collagen.

Sulfuration resistance of five experimental Ag-Pd-Au-Cu alloys with low Pd content of 10 or 12%.

Commercial Ag-based alloy (46Ag-20Pd-12Au-20Cu alloy) is widely used in Japan as a casting alloy. As opposed to the commercial composition, we prepared five experimental Ag-based alloys with reduced Pd content of 10 or 12%, increased Au content of 20 to 30%, and reduced Cu content of 12 to 20%. We then evaluated their sulfuration resistance by analyzing cast specimen surfaces dipped in 0.1% Na2S solution with SEM/EPMA, TF-XRD, and XPS. It became evident that all alloys were susceptible to sulfuration in the segregated Ag-rich Pd-poor phases. The degree and speed of sulfuration, however, differed among the six alloys examined. In particular, one experimental alloy (46Ag-10Pd-30Au-12Cu) possessed a sulfuration resistance equal or superior to that of commercial Ag-based alloy, while the other four experimental alloys were inferior in sulfuration resistance. Based on the results of this study, we concluded that our newly developed 46Ag-10Pd-30Au-12Cu alloy could be employed as a new sulfuration-resistant Ag-based casting alloy--which is especially useful if the price of Pd is skyrocketing again.

Accumulation of element Ti in macrophage-like RAW264 cells cultured in medium with 1 ppm Ti and effects on cell viability, SOD production and TNF-alpha secretion.

The adverse effect of Ti on body-defense macrophage is not well understood. The aims of this study were twofold: (1) to examine the intracellular accumulation of Ti element; and (2) to measure the cell viability, superoxide dismutase (SOD) production, and TNF-alpha secretion of macrophage-like RAW264 cells cultured for two days in medium with 1 ppm Ti prepared from acidic ICP Ti standard solution. PIXE analysis showed that element Ti was accumulated up to 7.3 ppm in RAW264 cells when cultured in the medium with 1 ppm Ti. Further, RAW264 cells cultured in the medium with 1 ppm Ti exhibited cell viability of about 60%, SOD production of about 180%, and TNF-alpha secretion of about 170% relative to those of control cells cultured in the medium without Ti. It was speculated that phagocytosis of minute Ti-containing complex (mostly TiO2) by macrophage caused oxidative stress and inflammatory reaction, leading to cell proliferation arrest and increased production of SOD and TNF-alpha.

Effects of Ni2+ ions on cell viability and NO production of murine peritoneal exudate cells (macrophages) with and without lipopolysaccharide stimulation.

The purpose of this study was to clarify the cytotoxicity of Ni2+ ions against murine peritoneal exudate cells (PEC) (macrophages). First, we examined the cell viability of PEC with and without lipopolysaccharide (LPS) stimulation in culture media containing Ni2+ ions up to 1000 micromol/L. Results showed that the cytotoxicity of Ni2+ ions against PEC was dose-dependent and accelerated by LPS stimulation, especially in media with Ni2+ ions exceeding 100 micromol/L. Second, we measured the production of nitric oxide (NO) from PEC and found that LPS caused the PEC to produce abundant NO. However, high dose of Ni2+ ions at concentration more than 200 micromol/L hindered and inhibited NO production. These results pointed out that the cytotoxicity of Ni2+ ions against macrophages depended on both the Ni2+ ion concentration and the presence of bacteria with LPS. Further, NO--a killer of bacteria--was lost when LPS-stimulated macrophages were exposed to high dose of Ni2+ ions.

Nonlinear large-deflection analysis of orthodontic wires.

The purposes of this study were (1) to measure the nonlinear force-deflection behavior of selected orthodontic wires using a conventional tensile test, (2) to extend a mathematical model for simulating the force system produced by orthodontic wires based on the small-deflection linear theory to the large-deflection nonlinear theory, and (3) to examine the effects of the cross-section and mechanical properties of orthodontic wires on nonlinear characteristics. A method for extending a mathematical model for simulating the force system produced by orthodontic wires based on the small-deflection linear theory to the large-deflection nonlinear theory was established, and this can provide a clear view of the true nature of orthodontic wires. Furthermore, our results demonstrated that the nonlinear properties of orthodontic wires were affected more by the cross-sectional shape than by mechanical properties.

Micromorphological changes in resin-dentin bonds after 1 year of water storage.

The purpose of this study was to evaluate the degradation of resin-dentin bonds after 1 year of water storage. Resin-dentin-bonded specimens were prepared with the use of an adhesive resin system (One-Step: Bisco). Half of the experimental specimens were sectioned perpendicular to the adhesive interface to produce a beam (adhesive area: 0.9 mm(2)) before being stored in distilled water at 37 degrees C for 1 year. The remaining half of the bonded specimens were sectioned into beams of similar dimensions after 1 year of water storage. Additional bonded specimens that had been stored in water for 24 h before sectioning into beams were used as controls. The beams in the two experimental groups and the control group were subjected to microtensile bond testing. Fractography was performed on all fractured beams with the use of FE-SEM. There were significant (p <.05) differences in bond strength among the control specimens (55.9 +/- 12.9 MPa), specimens that had been sectioned into beams after water storage (68.9 +/- 18.6 MPa), and specimens that had been sectioned into beams before water storage (28.1 +/- 9.3 MPa). Fractography revealed that the resin material was gradually extracted from the periphery to the center portion of the beam. This probably accounted for the decrease in bond strength after 1 year of water storage.

A novel method for the three-dimensional (3-D) analysis of orthodontic tooth movement-calculation of rotation about and translation along the finite helical axis.

The purpose of this study was to establish a novel method for evaluating orthodontic tooth movement in three-dimensional (3-D) space. The present system consisted of the following procedures at a given treatment period: (1) 3-D tooth positions were measured with a 3-D surface-scanning system using a slit laser beam; (2) the 3-D shape data were registered automatically at the maxillary first molars, and the coordinate systems were normalized; (3) the rotation matrix and translation vector were calculated from the automatic registration of the two position data for a given tooth; (4) the finite helical axes of teeth were calculated as the locus of zero rotational displacement; and (5) tooth movement was presented as rotation about and translation along the finite helical axis. To test this system, a male patient (age 22 yr 2 months) with Angle Class III malocclusion and moderate crowding of the anterior teeth, who had been treated using a standard multi-bracket appliance, was used as a model case in this study. Impressions for a dental cast model were taken at five phases; immediately before and after application of the appliance, and 10 days, 1 month and 2 months after beginning treatment. The results demonstrated that the present analytical method can more simply describe the movement of a given tooth by rotation about and translation along the finite helical axis, and provides quantitative visual 3-D information on complicated tooth movement during orthodontic treatment.