Biomedical subjects

Takashi Miyazaki

Publications and source records attributed to Takashi Miyazaki.

At least 19 recordsLinked to original sources

PubMed2006

In vivo performance of two different hydroxyapatite coatings on titanium prepared by discharging in electrolytes.

This study reports a discharging method for bone-like carbonated HA (cHA)-coating (Ca/P 1.71) and stoichiometric HA (sHA)-coating (Ca/P 1.67) with micrometer order thicknesses on titanium plates, using modified body fluid and acidic calcium phosphate solutions, respectively. In vivo histological performance of the HA coatings prepared by discharging in electrolytes was evaluated. Bone-contact indexes of HA coatings were measured microscopically. Additionally, bone-coating interface was analyzed by scanning electron microscopy and the use of an electron probe microanalyzer. Results demonstrated that there was no significant difference in contact index between HA coatings. However, the cHA coating was practically replaced by immature bone, and the titanium metal substrate was directly connected to the bone structure whereas the sHA coating layer remained and was partially detached from the titanium metal substrate. Since detached coating particles are pathogens, and can cause peri-implantitis, the cHA coating was more favorable than the sHA coating even if contact index was equivalent to that of the sHA coating. It is thought that coating thickness and chemical composition of coatings are important for biological stability of implants. In conclusion, since bone-like thin cHA coating showed high osteoconductivity and bone replacement, bone-like HA is superior to sHA coating for use in dental implants.

Coated Materials, Biocompatible↗

PubMed2006

Preliminary beta-tricalcium phosphate coating prepared by discharging in a modified body fluid enhances collagen immobilization onto titanium.

Because of its excellent scaffold properties toward bone cells, collagen has been recognized as a promising extracellular matrix protein for surface modification of titanium implants. Hydroxyapatite (HA) coatings have been investigated as a preliminary coating for collagen immobilization on titanium implants. However, the composition of HA-collagen is recognized as being difficult, and while many studies have suggested that biodegradable beta-tricalcium phosphate (beta-TCP) has better osteoconductivity than HA, the efficiency of preliminary beta-TCP coating for collagen immobilization on titanium surfaces has yet to be evaluated. This investigation aimed to evaluate the applicability of HA and beta-TCP coatings, prepared by discharging in modified body fluids, as preliminary collagen coatings. To increase collagen induction on preliminary HA and beta-TCP coatings, we used a new cathodic polarization method. X-ray photoelectron spectroscopy revealed that the bonding strength between the collagen NH(+) amino groups of collagen and phosphate (PO(4) (3-)) was greater on the beta-TCP coating than the HA coating. The preliminary beta-TCP coating was tightly crosslinked with RCOO(-) carboxyl groups of the collagen molecules and showed high cellular responses, even in the early stage of cell cultivation. Thus, this coating was found to be more effective than HA as a preliminary coating for collagen immobilization on titanium implants.

Animals↗

PubMed2006

Experimental binder-free investments reused to cast dental precious alloys.

This study aimed to develop reusable dental investments. SiO2 and MgO were selected as refractory materials to prepare three types of investment (coded as 60S-40M, 80S-20M, 100S) with 40, 20, and 0 wt% of MgO. Each type of investment was reused twice. Thermal expansion and compressive strength were examined and statistically evaluated by ANOVA. To evaluate fit of castings, full crowns were cast by using a commercial Au-Ag-Pd alloy with all investment types. Marginal fit was statistically analyzed by cement thickness. It was found that although MgO strengthened the mold, it had little influence on expansion. The strength of 60S-40M was the highest, and 100S had the greatest advantage with regard to thermal expansion. In the evaluation for clinical applicability, all investments were able to cast successfully, but their castings might be undersized. Among the experimental binder-free investments reused for dental casting in this study, 100S in particular showed to be a good candidate for repeated fabrication of precision fit castings.

Compressive Strength↗

PubMed2006

Simvastatin inhibits production of interleukin 6 (IL-6) and IL-8 and cell proliferation induced by tumor necrosis factor-alpha in fibroblast-like synoviocytes from patients with rheumatoid arthritis.

OBJECTIVE: Rheumatoid arthritis (RA) is a chronic inflammatory disease in which the synovial environment is characterized by intense immunological activity. Evidence suggests that statins modulate immune functions and may have a beneficial effect on patients with RA. We investigated whether simvastatin could inhibit the expression of interleukin 6 (IL-6) and IL-8 and cell proliferation induced by tumor necrosis factor-alpha (TNF-alpha) in fibroblast-like synoviocytes (FLS) obtained from RA patients undergoing joint replacement therapy. METHODS: RA FLS were cultured with or without 0.05-10 microM simvastatin for 12 h. Cytokine mRNA expression and secretion levels were detected using real-time PCR and ELISA, respectively. Cell proliferation of FLS induced by TNF-alpha was determined by MTT assay. RESULTS: Real-time PCR analysis revealed that the levels of IL-6 and IL-8 mRNA expressed by FLS were reduced by simvastatin in a dose-dependent manner. Levels of IL-6 and IL-8 in FLS culture supernatants were decreased by simvastatin in a time-dependent and dose-dependent manner. MTT assay revealed that simvastatin could inhibit proliferation of FLS induced by TNF-alpha. These effects of simvastatin on IL-6 and IL-8 production and cell proliferation were reversed in the presence of mevalonic acid or geranylgeranyl-pyrophosphate, but not with farnesyl-pyrophosphate. CONCLUSION: Our results suggest that the beneficial effect of simvastatin in RA patients may involve inhibition of IL-6 and IL-8 production, as well as reduction of cell proliferation.

Arthritis, Rheumatoid↗

PubMed2005

Novel method for titanium crown casting using a combination of wax patterns fabricated by a CAD/CAM system and a non-expanded investment.

OBJECTIVES: For titanium casting, most commercial investments for titanium recommend casting at a low mold temperature to reduce oxidation. However, the thermal expansion values of the molds at low casting temperatures may be insufficient. The purpose of the current study was to investigate the possibility of obtaining accurate titanium crown casts using wax pattern fabricated by a CAD/CAM system with a non-expanded mold. METHODS: Three types of experimental magnesia-based investments (A, B and C) were made and their properties were evaluated for dental use. Two kinds of wax patterns for full-coverage coping crowns (S-0: cement space of 0 microm; S-20: cement space of 20 microm) were fabricated using a commercial CAD/CAM system. A traditional method (TM) using inlay wax was performed for comparison. The investment for titanium casting was decided from the fundamental data of experimental investments. Titanium crowns were replaced on the stone die and the thickness of the cement layer was evaluated. RESULTS: There were no significant differences for the setting time and setting expansion among the experimental investments, but the aluminous cement content played a role in hardening and contracting the mold. The fit of the titanium crowns differed significantly between the TM and the CAD/CAM system. The ranges of thickness obtained from the TM, S-0 and S-20 were 20.78-357.88 microm, 25.12-107.46 microm and 17.84-58.92 microm, respectively. SIGNIFICANCE: High quality titanium crown casting was obtained using a combination of wax patterns fabricated by a CAD/CAM system and a non-expanded MgO-based investment.

Analysis of Variance↗

PubMed2005

Intracellular formation of collagen microfibrils in granulation tissue.

It is important to determine the biosynthesis process of collagen fibers to elucidate the mechanism by which granulation tissue is induced after injury. The purpose of this study is to investigate whether collagen microfibrils can be formed not only outside but also inside a cell. Fibroblast-like cells in granulation tissue resulting from incision and ligation were examined. The cells possessed vesicles containing collagen microfibrils. The vesicles were present in connection with Golgi apparatus or the rough endoplasmic reticulum. Furthermore, the vesicles were exhibited to be secretory granules with the secretory granule marker Rab3A. The fibroblast-like cells were also indicated to be myofibroblasts, using conventional transmission electron microscopy and immunoelectron microscopy for the myofibroblast marker alpha smooth muscle actin. In conclusion, it was demonstrated that collagen microfibrils could be formed in the cell in the case of collagen fiber overproduction.

Actins↗

PubMed2005

Cooperative structural change of actin filaments interacting with activated myosin motor domain, detected with copolymers of pyrene-labeled actin and acto-S1 chimera protein.

Acto-S1 chimera proteins CP24 and CP18 carry the entire actin sequence, inserted in loop 2 of the motor domain of Dictyostelium myosin II, and have MgATPase activity close to that of natural Dictyostelium actomyosin [M.S.P. Siddique, T. Miyazaki, E. Katayama, T.Q.P. Uyeda, M. Suzuki, Evidence against essential roles of subdomain 1 of actin in actomyosin sliding movements, Biochem. Biophys. Res. Commun. 332 (2005) 474-481]. Here, we examined and detected cooperative structural change of actin filaments accompanying interaction with myosin motor domain in the presence of ATP using copolymer filaments consisting of pyrene-labeled skeletal actin (SA) and either CP24 or CP18. Upon addition of ATP, the fluorescence intensity increased over the range from 380 to 480nm using 365-nm excitation. The relative increases of fluorescence intensity at 390nm were 14%, 46%, and 77% for the copolymer filaments with the CP24 to actin molar ratios of 0.0625, 0.143, and 0.333, respectively, and demonstrated a sigmoid behavior. Stoichiometric analysis indicates that each CP24 molecule appears to affect four actin molecules, on average, in SA-CP24 copolymers, and each CP18 molecule appears to affect three actin molecules in SA-CP18 copolymers.

Actin Cytoskeleton↗

PubMed2005

Evidence against essential roles for subdomain 1 of actin in actomyosin sliding movements.

We have engineered acto-S1chimera proteins carrying the entire actin inserted in loop 2 of the motor domain of Dictyostelium myosin II with 24 or 18 residue-linkers (CP24 and CP18, respectively). These proteins were capable of self-polymerization as well as copolymerization with skeletal actin and exhibited rigor-like structures. The MgATPase rate of CP24-skeletal actin copolymer was 1.06 s(-1), which is slightly less than the V(max) of Dictyostelium S1. Homopolymer filaments of skeletal actin, CP24, and CP18 moved at 4.7+/-0.6, 2.9+/-0.6, and 4.1+/-0.8 microm/s (mean+/-SD), respectively, on coverslips coated with skeletal myosin at 27 degrees C. Statistically thermodynamic considerations suggest that the S1 portion of chimera protein mostly resides on subdomain 1 (SD-1) of the actin portion even in the presence of ATP. This and the fact that filaments of CP18 with shorter linkers moved faster than CP24 filaments suggest that SD-1 might not be as essential as conventionally presumed for actomyosin sliding interactions.

Actins↗

PubMed2005

The Calvin cycle in cyanobacteria is regulated by CP12 via the NAD(H)/NADP(H) ratio under light/dark conditions.

In Synechococcus PCC7942 cells grown in the dark, the concentrations of NAD(H) and NADP(H) were 128+/-2.5 and 483+/-4.0 microm, respectively, while those in the cells under light conditions were 100+/-5.0 and 649+/-7.0 microm, respectively. Analysis of gel filtration indicated that the change of the ratio of NADP(H) to NAD(H) in cyanobacterial cells under light/dark conditions controls the reversible dissociation of the PRK/CP12/GAPDH complex (approximately 520 kDa) consisting of phosphoribulokinase (PRK), CP12, and glyceraldehyde-3-phosphate dehydrogenase (GAPDH). S. 7942 CP12 lacked the two Cys residues essential for formation of the N-terminal peptide loop in the CP12 of higher plants, but the N-terminal region of S. 7942 CP12 had the ability to be associated with PRK. The growth of mutant cells in which the CP12 gene was disrupted by a kanamycin resistance cartridge gene was almost the same as that of wild-type cells under continuous light conditions. However, under the light/dark cycle (12 h/12 h), the growth of CP12-disrupted mutant cells was significantly inhibited compared with that of wild-type cells. The mutant cells showed a decreased rate of O2 consumption and an increased level of ribulose 1,5-bisphosphate compared with wild-type cells in the dark. These data suggest that under light and dark conditions, the oligomerization of CP12 with PRK and GAPDH regulates the activities of both enzymes and thus the carbon flow from the Calvin cycle to the oxidative pentose phosphate cycle.

Amino Acid Sequence↗

PubMed2005

Colloidal beta-tricalcium phosphate prepared by discharge in a modified body fluid facilitates synthesis of collagen composites.

The development of hydroxyapatite/collagen composites that are naturally synthesized and need no additional treatment is required for use in bone repair. Since reducing the diameter can increase the specific surface area of calcium phosphate particles that can conjugate collagen molecules, we expected colloidal calcium phosphates of submicron diameter obtained by discharge to be effective in formulating these composites. Additionally, since biodegradable beta-tricalcium phosphate has better osteoconductivity than hydroxyapatite, this study aimed to investigate the synthesis of colloidal hydroxyapatite and beta-tricalcium phosphate/collagen composites. Collagen molecules were tightly polymerized in the beta-tricalcium phosphate/collagen composite by catalysis of the generated -P-O-P- polyphosphate chain. Bonding strength between collagen NH+ amino groups and -P-O-P-, and cross-linking of the Ca++-RCOO- in the collagen were increased compared with those in the hydroxyapatite/collagen composite. These chemical reactions due to colloidal beta-tricalcium phosphate might play a key role in the synthesis of collagen composites.

Biocompatible Materials↗

PubMed2005

Machining accuracy of CAD/CAM ceramic crowns fabricated with repeated machining using the same diamond bur.

The purpose of this study was to investigate the effect of repeated machining up to 51 times using the same diamond bur on machining accuracy of inner and outer surfaces of CAD/CAM (computer-aided designing and computer-aided manufacturing) machined ceramic crowns. The surface topography of machined crowns was examined using photographs. It was found that machining accuracy was not affected by the number of machining times. In all measuring points, the inner surface was machined to a dimension larger than the die model (i.e., increased gap), whereas the outer surface was machined to a dimension smaller than the crown model (i.e., smaller crown). Photo observation showed that cervical contour was machined in a clear, rounded form from 1st to 11th crowns.

Analysis of Variance↗

PubMed2005

Durability of diamond burs for the fabrication of ceramic crowns using dental CAD/CAM.

The diamond burs of two dental CAD/CAM systems (GN and CD) were examined if they could be used to fabricate up to 21 ceramic full crowns without fracture. After one, 11, and 21 machining times, the surfaces of the diamond burs were observed and the number of particles captured on SEM pictures was counted. The average surface roughness of the crowns was also measured. All diamond burs could be used to fabricate 21 ceramic crowns without fracture. A significant decrease in the number of diamond particles was found on the surfaces of GN burs after 11 and 21 machining times, but not on those of CD burs. The average surface roughness of GN crowns significantly increased with increase in the number of machining times. A significant positive correlation was found between the average surface roughness and the number of diamond particles.

Computer-Aided Design↗

PubMed2005

Recycling of used commercial phosphate-bonded investments with additional mono-ammonium phosphate.

Industrial waste is on the increase, resulting in severe environmental contamination. Against this background, this study sought to investigate the possibility of recycling used phosphate-bonded investments. Commercial phosphate-bonded investment was mixed and heated according to manufacturer's instructions, then powdered with a ball mill machine to be used as recycled investment (Code: R). Mono-ammonium phosphate (NH4H2PO4) of 2, 5, 8, 11, and 14 g were added to 100 g of R because of residual MgO, and coded as Ra, Rb, Rc, Rd, and Re respectively. Investment properties were then examined. Particle size of R was 50.66 microm and close to the original investment. It was found that more NH4H2PO4 led to larger setting expansion, smaller thermal expansion, and lower fired strength. The data of Re in these three aspects were 1.42 +/- 0.09%, 0.78 +/- 0.21%, and 8.85 +/- 0.88 MPa respectively, and differences against the original investment were significant. Ra showed significantly longer setting time than other investments. Based on the results of this study, we concluded that it was possible to set used phosphate-bonded investments by adding NH4H2PO4; in particular, Rd showed suitable expansion and acceptable mechanical properties.

Conservation of Natural Resources↗

PubMed2005

Fracture toughness measurement of dental ceramics using the indentation fracture method with different formulas.

This study examined fracture toughness (KIC) measurements obtained using the indentation fracture (IF) method with a view to improving their reliability. The KIC values of five dental ceramics were measured using the IF method with five different formulas, and the single-edge precracked beam (SEPB) method was used as a control. The elastic moduli of the dental ceramics were evaluated by dynamic hardness test. Load conditions of the dental ceramics that produced a median/radial crack for the IF method formulas were investigated. Based on the resultant c/a and P/c1.5 values, the indentation load (P) required for median/radial crack occurrence varied greatly from 29.4 to 196 N depending on the ceramic used. Among the five formulas, none of the KIC values obtained by the IF method with Miyoshi's formula differed significantly (p > 0.05) from the values obtained using SEPB method. These results suggested that, after an appropriate indentation load is determined, reliable KIC values for small dental ceramic specimens can be easily obtained using the IF method if Miyoshi's formula is used in combination with the dynamic hardness test.

Algorithms↗

PubMed2004

Changes in receptor activator of nuclear factor-kappaB, and its ligand, osteoprotegerin, bone-type alkaline phosphatase, and tartrate-resistant acid phosphatase in ovariectomized rats.

We investigated time-course changes in the expression of receptor activator of nuclear factor-kappaB (RANK), its ligand (RANKL), osteoprotegerin (OPG), bone-type alkaline phosphatase (BAP), and tartrate-resistant acid phosphatase (TRAP) in ovariectomized (OVX) rats. Samples of sera and coccyges were used for analysis of the enzyme activities and expression levels of proteins and mRNAs, and an immunohistochemical analysis was also performed. Serum BAP activity increased to 158.6% of the pre-operation value at 1 week after OVX, and then decreased to 38.7% at 8 weeks after OVX. On the other hand, the serum TRAP activity increased to 130.9% of the pre-operation level at 1 week after OVX, and was maintained at a high level, compared with the pre-operation level. The patterns of BAP and TRAP activity in the coccyges specimens were similar to those seen in the sera. The expression profiles of TRAP, RANK, and RANKL proteins in the coccyx specimens were similar to the pattern of serum TRAP activity, while the profiles of the BAP and OPG proteins were similar to the pattern of serum BAP activity in OVX rats. The changes in the mRNA expression levels of the osteogenic proteins were similar to those for protein expression. These biochemical changes in OVX rats were confirmed by immunohistochemical studies. Our results suggest that not only osteoclastogenesis accelerated but also osteoblastogenesis transiently increased during the early phase of osteoporosis.

Acid Phosphatase↗

PubMed2004

Myosin-induced volume increase of the hyper-mobile water surrounding actin filaments.

Microwave dielectric spectroscopy can measure the rotational mobility of water molecules that hydrate proteins and the hydration-shell volume. Using this technique, we have recently shown that apart from typical hydrating water molecules with lowered mobility there are other water molecules around the actin filaments (F-actin) which have a much higher mobility than that of bulk water [Biophys. J. 85 (2003) 3154]. We report here that the volume of this water component (hyper-mobile water) markedly increases without significant change of the volume of the ordinary hydration shell when the myosin motor-domain (S1, myosin subfragment-1) binds to F-actin. No hyper-mobile component was found in the hydration shell of S1 itself. The present results strongly suggest that the solvent space around S1 bound to F-actin is diffusionally asymmetric, which supports our model of force generation by actomyosin proposed previously [op. cit.].

Actin Cytoskeleton↗

PubMed2004

Glow discharge plasma pretreatment enhances osteoclast differentiation and survival on titanium plates.

Despite the fact that several reports have demonstrated osteoclast activity on various bioactive ceramics, osteoclast functions on surface-modified titanium have not come under focus. This study aimed to examine whether the increasing surface energy of glow discharge plasma (GDP) involved in protein adhesion containing the RGD (Arg-Gly-Asp) sequence affects osteoclast responses on titanium plates. We examined osteoclast differentiation and survival rates on titanium plates with and without GDP. The amounts of osteoclasts on titanium plates were not increased by GDP after 1 week. However, osteoclast differentiation was greatly activated by GDP pretreatment, as tartrate-resistant acid phosphatase synthesis significantly increased on the titanium plates with GDP. Additionally, since the presence of osteoclasts was detected only on the titanium plates with GDP, even after 4h cultivation in a coculture test, the osteoclasts survival rate was increased by GDP pretreatment. As osteoclast responses were affected even on surface modified metallic materials, we concluded that novel approaches are needed not only for osteoclastic resorption on ceramic materials but also for osteoclast responses on surface-modified metallic materials.

Acid Phosphatase↗

PubMed2004

alpha-Aminoadipate aminotransferase from an extremely thermophilic bacterium, Thermus thermophilus.

The extremely thermophilic bacterium Thermus thermophilus HB27 synthesizes lysine through alpha-aminoadipate (AAA). In this study, a T. thermophilus gene encoding the enzyme that catalyses transamination of AAA was cloned as a mammalian kynurenine/AAA aminotransferase (Kat2) gene homologue. A T. thermophilus mutant with disruption of the Kat2 homologue required a longer lag phase for growth and showed slower growth in minimal medium. Furthermore, addition of AAA or lysine shortened the lag phase and improved the growth rate. The Kat2 homologue was therefore termed lysN. LysN recognizes not only 2-oxoadipate, an intermediate of lysine biosynthesis, but also 2-oxoisocaproate, 2-oxoisovalerate and 2-oxo-3-methylvalerate, intermediates of leucine, valine and isoleucine biosyntheses, respectively, along with oxaloacetate, a compound in the TCA cycle, as an amino acceptor. These results suggest multiple roles of LysN in several cellular metabolic pathways including lysine and branched-chain amino acid biosyntheses.

2-Aminoadipate Transaminase↗