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Biomedical subjects

Tohru Hayakawa

Publications and source records attributed to Tohru Hayakawa.

At least 19 recordsLinked to original sources

Thermal ablation of FEL irradiation using gypsum as an indicator.

Thermal effects produced in a laser-irradiated sample were studied by micro-X-ray diffraction and micro-Fourier transform infrared spectroscopy (FTIR). Gypsum, transformed into bassanite at 124 degrees C and into anhydrite at 147 degrees C, was used as a thermal indicator. Pit formation by a wavelength-tunable free electron laser (FEL) irradiation on the gypsum pellet maximized at a wavelength of 3.0 microm, 2 mJ/shot, and pits were not detected in those irradiated at 2.6 or 3.8 microm compared with the maximum at 3.0 microm and diminished at 2.0 or 4.0 microm in the human tooth case. Micro-X-ray diffraction and micro-FTIR did not reveal any appreciable bassanite or anhydrite in the irradiated regions. From the laser ablation viewpoint, these results allow the FEL ablation to be considered as plasma or evaporative ones. This study indicated that the micro-pulse of laser was effective to prevent thermal damages of laser irradiation.

Calcium Sulfate↗

Characterization of sintered TCP sheets with various contents of binder prepared by tape-casting technique.

OBJECTIVES: The purpose of this study was to investigate the effect of binder content on the characterization of sintered tricalcium phosphate (TCP) sheets prepared by the tape-casting technique. METHODS: Poly(acrylic acid)-based binder was added at four contents: 5, 10, 15, or 30 mass%, to a beta-TCP aqueous slurry. The TCP green sheets were fabricated with the prepared TCP slurry by the tape-casting technique, and were sintered at 1200 degrees C for 2h. The sintered TCP sheets were characterized by field-emission scanning electron microscope (FE-SEM) observation, X-ray diffraction (XRD) and Fourier transform infrared (FT-IR) spectrometry. Apatite precipitation on the sintered TCP sheet surface in Hanks' balanced salt (HBS) solution at pH 7.4 was monitored over 14 days at 37 degrees C. RESULTS: A TCP green sheet with 5 mass% binder content had insufficient flexibility before sintering, and that with 30 mass% binder content had a problem in controlling the viscosity of slurry. FE-SEM observation showed that no differences in grain size existed in the various sintered TCP sheets. Measurement of XRD and FT-IR showed the presence of alpha-TCP phase in the various sintered TCP sheets, and same crystallographic structure was observed among various sintered TCP sheets. After 14 days immersion in HBS solution, the surfaces of sintered TCP sheets were completely covered with precipitated apatite crystals. SIGNIFICANCE: The binder content of 10 or 15 mass% is suitable for sintering TCP sheets. The sintered TCP sheet prepared by the tape-casting technique is a promising material as bone substitute.

Acrylic Resins↗

Pronase digestion of brush border membrane-bound Cry1Aa shows that almost the whole activated Cry1Aa molecule penetrates into the membrane.

Bacillus thuringiensis insecticidal proteins, Cry toxins, following ingestion by insect larvae, induce insecticidal effect by penetrating the brush border membranes (BBM) of midgut epithelial cells. Purified, activated B. thuringiensis Cry1Aa bound to Bombyx mori BBMV or unbound Cry1Aa were vigorously digested with Pronase. Both digests were compared by Western blotting. Free Cry1Aa was digested to alpha-helix and/or to amino acids at 1 mg Pronase/mL within 2.4 h at 37 degrees C. Whereas, BBMV-bound Cry1Aa was very resistant to Pronase digestion and even at 2 mg for 24 h, 7.5 kDa and approximately 30 kDa peptide were detected by alpha-2,3 antiserum, and alpha-4,5 and alpha-6,7 antisera, respectively. Another approximately 30 kDa peptide was also detected by beta-6-11 and domain III antisera. These fragments are believed either to be embedded in or to strongly interact with the BBMV. The 7.5 and former approximately 30 kDa peptides are thought to be derived from alpha-2,3 helix and stretch of alpha-4 to alpha-7 helices. Furthermore the latter approximately 30 kDa was thought to include the stretch of beta-6 to domain III. Moreover, the embedded Cry1Aa molecule appears to be segregated in some areas of beta-1-5 sheets, resulting in the above two approximately 30 kDa peptides. From these digestion patterns, we proposed new membrane insertion model for single Cry1Aa molecule. On the other hand, in digestion of BBMV-bound Cry1Aa, 15 kDa peptide which was recognized only by alpha-4,5 antiserum was observed. This fragment must be dimeric alpha-4,5 helices and we discussed the origin of this peptide.

Animals↗

Characterization and bioactivity of tape-cast and sintered TCP sheets.

The purpose of this study was to investigate the influence of sintering temperature on in vivo and in vitro bioactivities of sintered tricalcium phosphate (TCP) sheets prepared by the tape casting technique. Green sheets of beta-TCP prepared by tape casting were sintered for 2 h in a furnace at atmospheric pressure, at five different sintering temperatures: 900, 1000, 1100, 1150, and 1200 degrees C. Measurement of X-ray diffraction and Fourier transform infrared spectrometry showed the presence of alpha-tricalcium phosphate phase in the TCP sheet sintered at 1200 degrees C, in addition to beta-TCP phase. As compared with the other sintered TCP sheet, the TCP sheet sintered at 1200 degrees C demonstrated a lower dissolution rate in phosphate buffered saline solution at 37 degrees C and pH 7.4 over 24 weeks, and more amount of apatite formation in Hanks' balanced salt solution with pH 7.4 was observed. After 4 weeks' implantation of sintered TCP sheets into tibial diaphyses of rabbits, the bone-sheet contact of the TCP sheet sintered at 1200 degrees C was significantly higher than that of the TCP sheet sintered at 900 degrees C. These results indicate that a flat sintered TCP sheet prepared by tape casting is a promising material for a bone substitute.

Animals↗

Fibronectin immobilization using water-soluble carbodiimide on poly-L-lactic acid for enhancing initial fibroblast attachment.

The aim of this study is to evaluate the influence of fibronectin immobilization on poly-L-lactic acid (PLA) films on the initial attachment of human gingival fibroblasts. Carboxylic acid groups are chemically introduced on the PLA films' surface by surface hydrolysis with 0.5 M NaOH. The contact angle of PLA surface with respect to double-distilled water decreases significantly after NaOH hydrolysis. X-ray photoelectron spectroscopy (XPS) also reveals significantly higher intensities of C(C=O)/C(C-O) after NaOH hydrolysis. Fibronectin is immobilized onto the hydrolyzed PLA surface through a condensation reaction between the carboxylic acid groups on the hydrolyzed PLA surface and the amino groups of fibronectin using water-soluble carbodiimide. XPS analysis shows that the fibronectin-immobilized PLA surface is enriched with nitrogen atoms. The immobilization of fibronectin significantly enhances the number of initially attached human gingival fibroblasts on the PLA surface. No obvious differences in morphology are noted between fibroblasts cultured on native PLA and on fibronectin-immobilized PLA. Fibronectin can be immobilized onto the PLA surface after NaOH hydrolysis and this is effective in enhancing the initial attachment of human gingival fibroblasts.

Adsorption↗

Buffer solution can control the porosity of DNA-chitosan complexes.

The current studies examine the pore properties and biological effects of DNA-chitosan complexes, which may be useful as scaffolds for tissue engineering. The porosity of the DNA-chitosan complexes was controlled by rinsing them with several different pH 7.2 buffer solutions, including phosphate-buffered saline (PBS), Tris-HCl, boric acid, and N-(2-hydroxyethyl)piperazine-N'-(2-ethanesufonic acid) (HEPES). Rinsing with PBS resulted in 84% porosity, whereas rinsing with Tris-HCl produced 94% porosity. It was further found that daunorubicin hydrochloride complex intercalated with and bound to the groove of the DNA-chitosan complexes, indicating that DNA in the complexes maintains its double-stranded helical structure. The DNA-chitosan complexes were not toxic to MG-63 osteoblast-like cells and caused only a mild tissue response when implanted subcutaneously in the backs of rats. These results suggest that buffer-rinsed DNA-chitosan complexes may be useful as a scaffold material in tissue engineering.

Buffers↗

Adsorption behavior of antimicrobial peptide histatin 5 on PMMA.

Adsorption of antimicrobial peptide histatin 5 on a poly(methyl methacrylate) denture base may serve to prevent biofilm formation, leading to a reduction of denture-induced stomatitis. This study focused on adsorption behavior of histatin 5 onto PMMA surfaces modified using a cold plasma technique and the effectiveness of histatin 5 adsorption for reducing Candida albicans biofilm formation by the quartz crystal microbalance-dissipation (QCM-D) technique. PMMA spin-coated specimens were treated with oxygen (O(2)) plasma using a plasma surface modification apparatus. The amount of histatin 5 adsorbed onto the PMMA treated with O(2) plasma is more than six times greater than that adsorbed onto untreated PMMA. The degree of histatin 5 adsorption had a negative correlation with the contact angle, whereas that of zeta-potential showed no significant correlation. XPS analysis revealed that the introduction of the carboxyl and O(2) functional groups were observable on the O(2) plasma-treated PMMA. Increased surface hydrophilicity and the formation of the carboxyl could be responsible for histatin 5 adsorption on plasma-treated PMMA. There is no significant difference between histatin-adsorbed PMMA and control PMMA for C. albicans initially attached. On the contrary, the amount of C. albicans colonization on histatin-adsorbed PMMA was significantly less than the control.

Adsorption↗

Shear bond strength of calcium phosphate ceramic brackets to human enamel.

The aim of this study was to examine the shear bond strength between Hyaline brackets, a new type of calcium phosphate ceramic bracket, and human enamel using various types of adhesive resin and to investigate the effectiveness of a silane-coupling agent to bond Hyaline to human enamel. Kurasper F, Light Bond, Super Bond C&B, and Transbond XT were used as adhesive resins, and Porcelain Liner M was used as the silane-coupling agent. The Hyaline bracket was bonded to human enamel using one of the above adhesive resins according to the manufacturer's instructions. After applying the Porcelain Liner M to Hyaline, the Hyaline bracket was also bonded to enamel using one of the above adhesive resins according to the manufacturer's instructions. The shear bond strengths were measured after immersion in water at 37 degrees C for 24 hours. Three types of adhesive resin, Kurasper F, Light Bond, and Super Bond C&B, produced clinically acceptable shear bond strength with and without Porcelain Liner M. Transbond XT produced significantly lower bond strength to enamel with or without Porcelain Liner M (P < .05). The application of Porcelain Liner M was not useful for improving the bond strength of Hyaline to enamel. The adhesive remnant indices were not significantly different among four adhesive resins. In conclusion, adhesive resins such as Kurasper F, Light bond, and Super Bond C&B are useful for bonding esthetic Hyaline brackets to human enamel.

Analysis of Variance↗

A novel 96-kDa aminopeptidase localized on epithelial cell membranes of Bombyx mori midgut, which binds to Cry1Ac toxin of Bacillus thuringiensis.

Proteins in the brush border membrane (BBM) of the midgut binding to the insecticidal Cry1Ac toxin from Bacillus thuringiensis were investigated to examine the lower sensitivity of Bombyx mori to Cry1Ac, and new aminopeptidase N that bound to Cry1Ac was discovered. DEAE chromatography of Triton X-100-soluble BBM proteins from the midgut revealed 96-kDa aminopeptidase that bound to Cry1Ac. The enzyme was purified to homogeneity and estimated to be a 96.4-kDa molecule on a silver-stained SDS-PAGE gel. However, the native protein was eluted as a single peak corresponding to approximately 190-kDa on gel filtration and gave a single band on native PAGE. The enzyme was determined to be an aminopeptidase N (APN96) from its substrate specificity. Antiserum to class 3 B. mori APN (BmAPN3) recognized APN96, but peptide mass fingerprinting revealed that 54% of the amino acids of matched peptides were identical to those of BmAPN3, suggesting that APN96 was a novel isoform of the APN3 family. On ligand blots, APN96 bound to Cry1Ac but not Cry1Aa or Cry1Ab, and the interaction was inhibited by GalNAc. K(D) of the APN96-Cry1Ac interaction was determined to be 1.83 +/- 0.95 microM. The lectin binding assay suggested that APN96 had an N-linked bi-antennal oligosaccharide or an O-linked mucin type one. The role of APN96 was discussed in relation to the insensitivity of B. mori to Cry1Ac.

Amino Acid Sequence↗

Oxygen plasma surface modification enhances immobilization of simvastatin acid.

Simvastatin acid (SVA) has been reported to stimulate bone formation with increased expression of BMP-2. Therefore, immobilization of SVA onto dental implants is expected to promote osteogenesis at the bone tissue/implant interface. The aim of this study was to evaluate the immobilization behavior of SVA onto titanium (Ti), O(2)-plasma treated titanium (Ti + O(2)), thin-film coatings of hexamethyldisiloxane (HMDSO), and O(2)-plasma treated HMDSO (HMDSO + O(2)) by using the quartz crystal microbalance-dissipation (QCM-D) technique. HMDSO surfaces were activated by the introduction of an OH group and/or O(2)-functional groups by O(2)-plasma treatment. In contrast, titanium surfaces showed no appreciable compositional changes by O(2)-plasma treatment. The QCM-D technique enabled evaluation even at the adsorption behavior of a substance with a low molecular weight such as simvastatin. The largest amount of SVA was adsorbed on O(2)-plasma treated HMDSO surfaces compared to untreated titanium, HMDSO-coated titanium, and O(2)-plasma treated titanium. These findings suggested that the adsorption of SVA was enhanced on more hydrophilic surfaces concomitant with the presence of an OH group and/or O(2)-functional group resulting from the O(2)-plasma treatment, and that an organic film of HMDSO followed by O(2)-plasma treatment is a promising method for the adsorption of SVA in dental implant systems.

Adsorption↗

Non-linear finite element analysis of the failure progression of fiber-reinforced ceramics produced by tape casting technique.

The purpose of this study was to investigate the failure progression process of fiber-reinforced ceramic by finite element (FE) analysis. The three-dimensional FE model for three-point bending simulation was 40 mm long, 4 mm wide, 3 mm thick, and with a span length of 30 mm. Nodal force with load increment of 20 N was applied at the center of the upper surface of the beam. To evaluate matrix fracture and fiber fracture, von Mises criterion and Tsai-Hill criterion were used respectively. Consequently, the stress-deflection curve obtained from FE simulation agreed with that obtained from the experimental testing. Differences in flexural strength and modulus between the analytical and experimental results were 1.3 and -2.9% respectively--demonstrating a close agreement between both results. In conclusion, the FE model applied in the present study was shown to be valid for predicting the failure progression of fiber-reinforced ceramics.

Aluminum Oxide↗

Mode superposition transient dynamic analysis for dental implants with stress-absorbing elements: a finite element analysis.

The purpose of this study was to analyze the dynamic behavior of a dental implant with a stress-absorbing element, using dynamic analysis. Two model types, stress-absorbing model with a resilient stress absorber made of polyoxymethylene and non-stress-absorbing model with rigid titanium, were employed. In both model types, the implant was 4.0 mm in diameter and 13.0 mm in length and placed in the mandibular first molar region. Shapes of the finite element implant and implant-bone were modeled using computer-aided design. All calculations for the dynamic analysis were performed using the finite element method. It was found that the stress-absorbing model had a lower natural frequency than the non-stress-absorbing model. In addition, the stress-absorbing model had a higher damping effect than the non-stress-absorbing model. It was concluded that mode superposition transient dynamic analysis is a useful technique for determining dynamic behavior around dental implants.

Dental Implants↗

Modification of 4-META/MMA-TBB resin for safe debonding of orthodontic brackets--influence of the addition of degradable additives or fluoride compound.

The purpose of this study was to evaluate the performance of modified 4-META/MMA-TBB resin cements (Superbond C&B) in terms of debonding orthodontic brackets easily and safely from enamel without the loss of proper bracket bond strength. Poly (DL-lactide-co-glycolide) (PLGA), calcium fluoride (CaF2), or alpha-tricalcium phosphate (alpha-TCP) was added to the polymer powder of 4-META/MMA-TBB resin, and the shear bond strength of orthodontic brackets to human enamel using modified resins was measured before and after 10,000-cycle thermal cycling test between 5 degrees C and 55 degrees C. The modified resins tended to provide lower bond strength compared with the original 4-META/MMA-TBB resin. However, alpha-TCP- or CaF2-modified resin showed no significant differences in bond strength before and after thermal cycling. Moreover, there was a tendency of more residual resin remaining on the tooth surface after debonding, thereby suggesting a lower risk of enamel fracture. In conclusion, alpha-TCP- or CaF2-modified 4-META/MMA-TBB resin seemed to allow easy and safe debonding of orthodontic brackets without loss of proper bracket bond strength.

Acid Etching, Dental↗

Fabrication, characterization, and biological assessment of multilayered DNA-coatings for biomaterial purposes.

This study describes the fabrication of two types of multilayered coatings onto titanium by electrostatic self-assembly (ESA), using deoxyribosenucleic acid (DNA) as the anionic polyelectrolyte and poly-d-lysine (PDL) or poly(allylamine hydrochloride) (PAH) as the cationic polyelectrolyte. Both coatings were characterized using UV-vis spectrophotometry, atomic force microscopy (AFM), X-ray photospectroscopy (XPS), contact angle measurements, Fourier transform infrared spectroscopy (FTIR), and for the amount of DNA immobilized. The mutagenicity of the constituents of the coatings was assessed. Titanium substrates with or without multilayered DNA-coatings were used in cell culture experiments to study cell proliferation, viability, and morphology. Results of UV-vis spectrophotometry, AFM, and contact angle measurements clearly indicated the progressive build-up of the multilayered coatings. Furthermore, AFM and XPS data showed a more uniform build-up and morphology of [PDL/DNA]-coatings compared to [PAH/DNA]-coatings. DNA-immobilization into both coatings was linear, and approximated 3microg/cm(2) into each double-layer. The surface morphology of both types of multilayered DNA-coatings showed elevations in the nanoscale range. No mutagenic effects of DNA, PDL, or PAH were detected, and cell viability and morphology were not affected by the presence of either type of multilayered DNA-coating. Still, the results of the proliferation assay revealed an increased proliferation of primary rat dermal fibroblasts on both types of multilayered DNA-coatings compared to non-coated controls. The biocompatibility and functionalization of the coatings produced here, will be assessed in subsequent cell culture and animal-implantation studies.

Animals↗

Analysis of photopolymerization behavior of UDMA/TEGDMA resin mixture and its composite by differential scanning calorimetry.

The aim of this study was to investigate the extent of polymerization (Ep) in terms of polymerization rate of UDMA/TEGDMA resin mixtures and its composite resin, by using a differential scanning calorimeter (DSC) technique employing a photopolymerization apparatus. The resin mixtures used in this study consisted of urethane dimethacrylate (UDMA) as a base monomer and triethyleneglycol dimethacrylate (TEGDMA) as a low viscosity monomer. The concentration of TEGDMA in the mixed monomer was varied to 0, 20, 40, 60, 80, and 100 mol %. Additionally, using a base monomer consisting of 60 mol % UDMA and 40 mol % TEGDMA, four kinds of composites with silica filler of 0, 20, 40, 60, and 70 wt %, were prepared in this study. The general reaction profile and Ep values were obtained from the DSC curves. Increasing the concentration of TEGDMA resulted in a decrease in the viscosity of the UDMA/TEGDMA mixture, a delay in the time to maximum polymerization rate, and an increase in the Ep values of the resin mixtures. Furthermore, Ep values decreased with increasing filler content between 0 and 60 wt % but did not decrease further between 60 and 70 wt %.

Calorimetry, Differential Scanning↗

Tape-casting technique can prepare beta-TCP sheets with uniform thickness and flexibility.

The objective of this study is to propose a new fabrication technology for bone substitutes. In this study, a tape-casting method was used to prepare flexible beta-tricalcium phosphate (beta-TCP) sheets. A beta-TCP slurry containing a binder and plasticizer was used in a doctor blade system. The beta-TCP sheet obtained by this tape-casting method was highly flexible, enabling twisting and free-form shaping. The beta-TCP sheet was approximately 0.21 mm thick. X-ray diffraction and Fourier transform infrared spectrometry revealed that the structure of the beta-TCP component in the sheet is the same as that of the original beta-TCP powder. Observation by field-emission scanning electron microscopy showed that the beta-TCP sheet had a flat, microgranular surface. During the early stages, the tensile stress-strain curves of the beta-TCP sheet showed a nonlinear behavior until reaching the point of final fracture. This result was derived from the ductile property of the prepared beta-TCP sheet. In conclusion, a flexible beta-TCP sheet was easily prepared using a tape-casting technique. Fabrication using tape casting offers the advantages of enabling the preparation of ceramic sheets with precise thickness and not requiring expensive fabrication facilities.

Biocompatible Materials↗

Bonding durability of using self-etching primer with 4-META/ MMA-TBB resin cement to bond orthodontic brackets.

This study determines the bonding durability when a self-etching primer is used with Superbond C&B (a 4-methacryloxyethyl trimellitate anhydride/methyl methacrylate-tri-n-butyl borane resin) to bond orthodontic brackets to enamel. Thermocycling test was used to assess bonding durability. Metal brackets were bonded to the phosphoric acid-etched or Megabond self-etching primer-treated enamel surface of human premolars using Superbond C&B. The shear bond strengths were measured after immersion in water at 37 degrees C for 24 hours or after 2000 or 5000 cycles of thermocycling between 5 degrees C and 55 degrees C. Data were analyzed using 2-way analysis of variance and Fisher's protected least significant difference test for multiple comparisons. There was no significant difference in shear bond strength between phosphoric acid and Megabond self-etching primer treatment before the thermocycling test. After 2000 and 5000 thermal cycles, significant decreases in shear bond strength were observed with phosphoric acid etching. On the contrary, no significant differences in shear bond strength were observed after 2000 and 5000 thermal cycles with Megabond self-etching primer. The adhesive remnant indices were not significantly different between phosphoric acid etching and Megabond self-etching primer treatment either before or after thermal cycles. This study suggested that when used with Superbond C&B in bonding orthodontic brackets, Megabond self-etching primer is superior to phosphoric acid as an enamel preparation agent in providing durable bond strength.

Acid Etching, Dental↗

Evaluation of data in terms of two-dimensional random walk model: interaction between NADH-cytochrome b5 reductase and cytochrome b5.

Normally, bimolecular reactions are analyzed in terms of the Smoluchowski theory. However, when one attempts to generalize this analysis to cases where diffusion proceeds in two other than in three dimensions, one soon encounters severe conceptual difficulties. Although kinetic studies of membrane enzymes are generally difficult because the usual kinetic formalism refers to nonaggregated homogenous solutions, a major goal of our research is to define the molecular mechanism(s) by which alterations in membrane-bound substrate contents affect the enzyme activity in the same membrane. For that purpose, a simplified random-walk model was adopted in the present work. The enzyme reaction in the two-dimensional membrane could be calculated theoretically by applying the classical analysis of heat equation. As a result, the theoretical rate equation well accounting experimental findings was derived on the model of the liver microsomal NADH-cytochrome b5 reductase reaction. Furthermore, it was found that the modification of the simple rigid-sphere collision theory by including a term called the steric factor was not necessary in this derived equation.

Animals↗