Biomedical subjects

T Kokubo

Publications and source records attributed to T Kokubo.

At least 145 records · Page 8Linked to original sources

PubMed1997

Bonding of chemically treated titanium implants to bone.

A study was undertaken in rabbit tibiae to determine the effects of chemical treatments and/or surface-induced bonelike apatite on the bone-bonding ability of titanium (Ti) implants. Smooth-surfaced plates (10 x 10 x 2 mm) of pure Ti, alkalil- and heat-treated Ti, and bonelike apatite-formed Ti after the treatments were implanted into the tibial metaphyses of mature rabbits. The tibiae containing the implants were harvested at 4, 8, and 16 weeks after implantation and subjected to a tensile testing and histologic evaluation. Biomechanical results showed that both treated implants exhibited significantly higher failure loads compared with untreated Ti implants at all time periods. Histologic examination by Giemsa surface staining, contact microradiography (CMR), and scanning electron microscopy (SEM) in backscatter mode revealed that both treated Ti implants directly bonded to bone tissue during the early postimplantation period, whereas untreated Ti implants formed direct contact with the bone only at 16 weeks. SEM-electron-probe microanalysis (EPMA) examination showed a Ca-P-rich layer at the interface between the treated implants and bone, although the Ca-P-rich layer was not detected on the surface of untreated implants during observation periods. The results of this study suggest that chemical treatments may accelerate the bone-bonding behavior of titanium implants and enhance the strength of bone-implant bonding by inducing a bioactive surface layer on Ti implants.

Animals↗

PubMed1997

Bonding strength of bonelike apatite layer to Ti metal substrate.

Our previous study showed that titanium metal forms a bonelike apatite layer on its surface in simulated body fluid when it was subjected to NaOH and heat treatments to form a sodium titanate hydrogel or amorphous sodium titanate surface layer. In the present study, bonding strength of the apatite layer formed on the titanium metals to the substrates were examined under tensile stress, in comparison with those of the apatite layers formed on Bioglass 45S5-type glass, dense sintered hydroxyapatite, and glass-ceramic A-W, which are already clinically used. The NaOH-treated titanium metals showed higher bonding strength of the apatite layer to the substrates, which was maximized by heat treatments at 500 and 600 degrees C, than all the examined bioactive ceramics. It is believed that bioactive metals thus obtained are useful as bone substitutes, even under load-bearing conditions.

Apatites↗

PubMed1997

Preparation of radiotherapy glass by phosphorus ion implantation at 100 keV.

A chemically durable glass containing a large amount of phosphorus is useful for in situ irradiation of cancers. It can be activated to be a beta emitter (half-life of 14.3 days) by neutron bombardment. Microspheres of the activated glass injected into the tumors can irradiate the tumors directly with beta rays without irradiating neighboring normal tissues. In the present study a P+ ion was implanted into a pure silica glass in a plate form at 100 keV in order to find the fundamental conditions for obtaining such a glass. Little phosphorus was present in the surface region, at least to a depth of 2.4 nm for doses of 5 x 10(16) and 1 x 10(17) cm-2, whereas an appreciable amount of it was distributed on the glass surface and a part of it was oxidized for doses above 5 x 10(17) cm-2. The glasses implanted with doses of 5 x 10(16) and 1 x 10(17) cm-2 hardly released the P and Si into water at 95 degrees C, even after 7 days, whereas the glasses implanted with doses above 5 x 10(17) cm-2 released appreciable amounts of these elements. Implantation energies of 20 and 50 keV (even at doses of 5 x 10(16) and 1 x 10(17) cm-2, respectively), formed oxidized phosphorus on the glass surfaces and gave appreciable release of the P and Si into the hot water. This indicates that a chemically durable glass containing a larger amount of phosphorus could be obtained if a P+ ion is implanted at higher energies to localize in a deeper region of the glass surface.

Biocompatible Materials↗

PubMed1997

Apatite layer-coated titanium for use as bone bonding implants.

For development of thin bioactive coatings on metal implants, a dense and uniform apatite layer was coated onto titanium (Ti) implants in situ by using a new biomimetic method, which is composed of apatite nucleation and growth steps in simulated body fluid (SBF). Analysis of the coatings by thin film X-ray diffraction and scanning electron microscopy-energy dispersive X-ray microanalysis (SEM-EMPA) before implantation showed that its characteristics were very similar to those of natural bone. The coated and uncoated rectangular plates were bilaterally implanted into the tibial proximal metaphyses of rabbits. After 6, 10 and 25 weeks post-implantation, the bone bonding and bone formation at the bone-implant interfaces were evaluated by a detachment test and undecalcified histological examination. Mechanical testing in tension showed that the failure load of apatite layer-coated Ti implants was significantly higher than that of uncoated control at each time period (all P < 0.001). Histologically, it was shown that bone was deposited directly onto the apatite coating without any intervening soft tissue, while in the paired controls, interpositional soft tissue was seen at the bone-implant interface. By SEM-EPMA, a uniform calcium- and phosphorus-rich layer was detected between the coated implants and bone, but not in uncoated controls at either earlier or later time periods. The results indicate that the apatite layer deposited on Ti in situ may significantly increase the bone bonding strength by providing a bioactive surface, which allows for an early bone apposition to the implant. In addition, the apatite layer-coated Ti produced by the biomimetic process may fulfil the requirements of favourable thin coatings and strong adhesion at the metal-coating interface.

Analysis of Variance↗

PubMed1997

Bioactivity of ferrimagnetic glass-ceramics in the system FeO-Fe2O3-CaO-SiO2.

Bioactive and ferrimagnetic glass-ceramics are useful as thermoseeds for hyperthermia treatment of cancer. A heat treatment of a 40(FeO, Fe2O3)-60CaO x SiO2 wt% glass gives a ferrimagnetic glass-ceramic containing 36 wt% magnetite in a CaO x SiO2 matrix. However, it does not show bioactivity since a small amount of iron ion remains in the matrix. In the present study, bioactivities of ferrimagnetic glass-ceramics which were prepared by heat treatment of glasses of the composition 40(FeO, Fe2O3)-60CaO x SiO2 (wt%) with various components added at 100:3 weight ratio were evaluated in vitro by examining bone-like apatite formation on their surfaces in a simulated body fluid. It was found that glass-ceramics with Na2O or B2O3 added in combination with P2O5 show bioactivity.

Apatites↗

PubMed1997

Induction of bioactivity of a non-bioactive glass-ceramic by a chemical treatment.

Glass-ceramic A-W(Al), which was prepared by heat treatment of a MgO-CaO-SiO2-P2O5-Al2O3 glass to precipitate crystalline apatite and wollastonite, shows a higher mechanical strength than glass-ceramic A-W, which was prepared by heat treatment of a MgO-CaO-SiO2-P2O5 glass to precipitate the same types of crystalline phases. The former, however, does not show bone-bonding ability, i.e. bioactivity, whereas the latter shows it. In the present study, in order to induce bioactivity of glass-ceramic A-W(Al), it was treated with HCl or NaOH solutions with different concentrations, and its bioactivity was evaluated by examining the apatite formation on its surface in a simulated body fluid (SBF) with ion concentrations nearly equal to those of human blood plasma. When the glass-ceramic A-W(Al) was pretreated with HCl aqueous solutions with concentrations over 0.1 M, it formed the bone-like apatite on its surface in SBF. This was attributed to the formation of a hydrated silica on its surface by the HCl treatment.

Apatites↗

PubMed1997

Antibiotic delivery system using bioactive bone cement consisting of Bis-GMA/TEGDMA resin and bioactive glass ceramics.

A novel drug delivery system containing cephalexin (CEX) as a model drug using a new bioactive bone cement consisting of 15% bisphenol-alpha-glycidyl methacrylate (Bis-GMA), 15% triethylene-glycol dimethacrylate (TEGDMA) resin and 70% apatite- and wollastonite-containing glass-ceramic (A-W GC) powder was investigated. A-W GC powder containing CEX powder hardened within 5 min after mixing with Bis-GMA/TEGDMA resin, and furthermore its compressive strength was expected to be higher than that of polymethylmethacrylate cement. In vitro CEX release from bioactive bone cement pellets in a simulated body fluid at pH 7.25 and 37 degrees C continued for more than 2 weeks. The drug release rate increased with increasing amount of CEX in the mixture. All of the drug release profiles followed the Higuchi equation at the initial stage, but not at later stages. As hydroxyapatite was precipitated out on the cement surface, the drug release rate decreased. These results suggest that the CEX release rate from bioactive bone cement could be controlled by varying the amount of drug in the cement system.

Biomechanical Phenomena↗

PubMed1997

Localization of cathepsin K in human osteoclasts by in situ hybridization and immunohistochemistry.

We have recently cloned cathepsin K from a human bone cDNA library. Since cathepsins are proposed to be involved in the degradation of mineralized bone matrix, we have investigated, by in situ hybridization and immunocytochemistry, the expression of the cathepsin K mRNA transcripts and protein in sections of bone and giant cell tumor to determine which cells express this enzyme. Within all tissues studied, cathepsin K was highly expressed in osteoclasts. Furthermore, the expression of cathepsin K mRNA in giant cell tumor tissue appeared to be confined to the periphery of the osteoclast indicating a compartmentalization of the mRNA. Immunohistochemistry confirmed the specific localization of cathepsin K to the osteoclast. In actively resorbing osteoclasts, the immunostaining was localized at the ruffled border, whereas in osteoclasts in sections of giant cell tumor, staining was observed in lysosomal vacuoles, which in some cases were seen to fuse with the cell membrane. Other cells within the bone, such as osteoblasts and osteocytes, did not express either the cathepsin K transcript or protein. However, there were very low levels of cathepsin K detected in a population of mononuclear cells, possibly representing osteoclast progenitor cells, within the marrow/stromal layer. The specific localization of cathepsin K within osteoclasts would therefore indicate the potential role of this enzyme in the bone resorptive process.

Blotting, Northern↗

PubMed1997

Effect of heat treatment on apatite-forming ability of Ti metal induced by alkali treatment.

The present authors previously showed that titanium metal forms a bone-like apatite layer on its surface in a simulated body fluid (SBF), when it has been treated with a NaOH solution to form a sodium titanate hydrogel layer on its surface. This indicates that the NaOH-treated Ti metal bonds to living bone. The gel layer as-formed is, however, mechanically unstable. In the present study, the NaOH-treated Ti metal was heat treated at various temperatures in order to convert the gel layer into a more mechanically stable layer. The gel layer was dehydrated and transformed into an amorphous sodium titanate layer at 400-500 degrees C, fairly densified at 600 degrees C and converted into crystalline sodium titanate and rutile above 700 degrees C. The induction period for the apatite formation on the NaOH-treated Ti metal in SBF increased with the transformation of the surface gel layer by the heat treatment. Ti metal heat treated at 600 degrees C, however, showed a fairly short induction period as well as high mechanical stability, since it was covered with a fairly densified amorphous layer.

Journal Article↗

PubMed1997

Epitope analysis of birch pollen allergen in Japanese subjects.

Birch pollen is a very common cause of nasal allergy (pollinosis) not only in Scandinavia, Europe, Canada, and the northern part of the United States but also in Hokkaido, Japan. We have previously reported a positive association between the HLA-DR9 phenotype and the development of birch pollen allergy in Japanese subjects. However, there is little information about T cell epitopes of birch pollen which are presented by HLA class II molecules other than HLA-DR9. Therefore, we analyzed the difference in T cell epitope usage in patients who had HLA-DR9 versus those who did not. Seven Japanese patients with birch pollinosis were studied. Some groups of peptides representing T cell epitopes (Betula verrucosa; Bet VI peptides, p7-33, p23-46, p138-160) appeared to be shared by the majority, while another peptide (Bet VI p72-95) was recognized predominantly by patients who expressed HLA-DR9 and/or HLA-DQ3 molecules. Moreover, seven T cell clones and eight T cell lines were generated from two patients who did not have HLA-DR9 or HLA-DQ3. Using some of these T cell clones/lines, we investigated the relationship between HLA class II molecules and antigenic peptides. One of these T cell clones recognized antigenic peptides in the context of the HLA-DQ1 molecule. To our knowledge, this is the first indication that the epitope on Bet VI can be presented by the HLA-DQ molecule.

Allergens↗

PubMed1997

Evidence for involvement of IgA1 hinge glycopeptide in the IgA1-IgA1 interaction in IgA nephropathy.

The study was performed to investigate the role of the IgA1 hinge region in the IgA1-IgA1 interaction, which was observed previously in IgA nephropathy. The competitive inhibition assays of the IgA1-IgA1 binding were performed using the following candidates for inhibitors: native IgA1 hinge glycopeptide (nHGP), IgA1, IgA2, and IgG. The IgA1-IgA1 binding was definitely inhibited by the nHGP and the IgA1 (maximum of percent inhibition: 66.1 and 60.5%, respectively). There was no obvious inhibition in the IgA2 and the IgG. The inhibition curves of the nHGP and the IgA1 were significantly different from that of the IgG (P < 0.01, respectively). Furthermore, to reveal the detailed binding sites in the interaction, the same inhibition assays were performed using the following substances composing the IgA1 hinge glycopeptide: galactose (Gal), N-acetyl-galactosamine (GalNAc), Gal beta 1-3GalNAc, sialic acid, tetrapeptide PTPS, and synthesized hinge proline-rich peptide PVPSTPPTPSPSTPPTPSPS (sHP). sHP, Gal beta 1-3GalNAc, Gal, and GalNAc inhibited the binding (69.3, 34.1, 14.9, 14.6%, respectively). No obvious inhibition was observed in sialic acid and tetrapeptide PTPS. The inhibition curve of sHP was significantly different from that of the PTPS (P < 0.05). Those of Gal beta 1-3GalNAc, Gal, and GalNAc were also significantly different from that of sialic acid (P < 0.05, respectively). These results suggested that the IgA1-IgA1 interaction could be mediated by the core structure including the peptide and the sugars, except for sialic acid in the hinge region, resulting in the formation of the circulating macromolecular IgA1 in IgA nephropathy.

Adult↗

PubMed1997

[Mediastinal lymph node metastasis from esophageal carcinoma: CT assessment with pathologic correlation].

Computed tomographic (CT) scans were performed in 179 patients with esophageal carcinoma to evaluate mediastinal lymph node metastasis. Histopathologic findings were compared with CT findings in a total of 7,218 resected lymph nodes. First, the criterion for lymph node metastasis on CT scans was 10 mm or more in long transverse diameter. The overall sensitivity and positive predictive value (PPV) were 19% (60 of 317 nodes) and 33% (60 of 180 nodes), respectively. Analysis of each of the eight subgroups of mediastinal nodes revealed that the PPV was more than 70% in node Nos. 105, 108, 110, and 112. In other subgroups, however, the PPV was less than 60%. Sensitivity was less than 50% in all eight subgroups. Second, the criterion for metastasis was 10 mm or more in short transverse diameter. The overall sensitivity and PPV were 8% (26 of 317 nodes) and 63% (26 of 41 nodes), respectively. Analysis of subgroups showed that the PPV in No. 106 nodes increased to 92%. In No. 106 nodes, use of a 5 mm criterion in long transverse diameter increased sensitivity to only 55%. Of the 317 histopathologically proven metastatic lymph nodes, 90 nodes (28%) were 10 mm or more in size, 112 (35%) were 5-10 mm, and 115 (36%) were less than 5 mm. Of the 6,901 non-metastatic lymph nodes, 473 nodes (7%) were 10 mm or more in size. Small (less than 5 mm in size) metastatic nodes were present in all eight subgroups. Among No. 107 and 109 nodes, large (10 mm or more in size) nonmetastatic nodes were prominent, resulting in low sensitivity and PPV. We conclude that CT does not provide an accurate assessment of metastatic versus non-metastatic mediastinal lymph nodes in patients with esophageal carcinoma.

Esophageal Neoplasms↗

PubMed1997

Bone bonding ability of bioactive bone cements.

The bone bonding ability of three types of bioactive bone cement A, B, and C consisting of glass or glass ceramic powder and bisphenol-alpha-glycidyl methacrylate resin was evaluated. Type A contained MgO-CaO-SiO2-P2O5-CaF2 glass powder; Type B, MgO-CaO-SiO2-P2O5-CaF2 glass ceramic powder; and Type C, MgO free CaO-SiO2-P2O5-CaF2 glass powder. Rectangular plates (2 x 10 x 15 mm) of Types A, B, C, and polymethylmethacrylate cements were implanted into the tibial metaphyses of male rabbits and the failure load measured by mechanical failure testing (detaching test) 10 and 25 weeks after implantation. The failure loads of Types A, B, C, and polymethylmethacrylate cements were respectively, 29.52, 41.48, 28.22, and 0.29 N at 10 weeks and 33.42, 41.27, 33.64, and 0.20 N at 25 weeks. Examination of the bone cement interface revealed that all the bioactive bone cements achieved direct bone contact with the bone. These results showed that all three types of bioactive bone cement have the ability to bond to bone, and the cement containing glass ceramic powder revealed higher bonding strength than did those containing glass powder.

Animals↗

PubMed1997

[Usefulness and limitations of 99mTc-MIBI scintigraphy for detection of parathyroid lesions].

To evaluate the usefulness of MIBI scintigraphy (MIBI) for parathyroid lesions, the detectability of lesions by MIBI was compared with that by Tl-Tc subtraction imaging, ultrasonography, CT and MRI in 56 histologically proved lesions. In neck lesions, ultrasonography (92%) and MIBI (85%) showed better detectability than those by the other three modalities. With MIBI, detectability was decreased for smaller parathyroid lesions that coexisted with thyroid disease. Among the five modalities, MIBI showed the highest detectability (88%) for ectopic or metastatic lesions. The smallest parathyroid lesions detected by MIBI were a parathyroid adenoma weighing 220mg and a parathyroid hyperplasia weighing 200mg. MIBI was thought to be more valuable for ectopic or metastatic parathyroid lesions.

Adenoma↗

PubMed1997

The in vitro and in vivo indomethacin release from self-setting bioactive glass bone cement.

The in vivo and in vitro drug release profiles from a self-setting bioactive CaO-SiO2-P2O5 glass bone cement containing indomethacin as a model drug were investigated. The cement containing 2% and 5% indomethacin (IMC) powder hardened within 5 min after mixing with ammonium phosphate buffer. After setting, in vitro drug release from drug-loaded cement pellets in a simulated body fluid (SBF) at pH 7.25 and 37 degrees C continued for two weeks. The hardened cement gradually formed low-crystallinity hydroxyapatite during the drug release test in SBF. An IMC-loaded cement device (2% and 5% drug) was implanted in the subcutaneous tissue on the back of rats. The in vivo IMC release from the cement increased and attained maximum levels (Cmax of 2% and 5% drug-loaded cements was 0.27 and 3.37 micrograms/ml, respectively) at Tmax, 3 and 0.5 d, respectively, upon subcutaneous (s.c.) administration in rats. This suggested that the s.c. administration of the cement provided IMC release for a much longer period than s.c. administration of the solution, and the plasma IMC concentration was dependent on the drug concentration in the cement. The plasma IMC concentration and the area under the curve from 2% and 5% IMC-loaded cements in rats were dependent on the concentration of IMC in the cements. The in vivo IMC concentration in plasma obtained by the deconvolution method was much lower than that delivered in SBF in vitro. Scanning electron microscopy and photomicrographs of cross sections showed that the bioactive bone cement had excellent biocompatibility with the surrounding soft tissues.

Animals↗

PubMed1996

The TAF(II)250 subunit of TFIID has histone acetyltransferase activity.

The transcription initiation factor TFIID is a multimeric protein complex composed of TATA box-binding protein (TBP) and many TBP-associated factors (TAF(II)s). TAF(II)s are important cofactors that mediate activated transcription by providing interaction sites for distinct activators. Here, we present evidence that human TAF(II)250 and its homologs in Drosophila and yeast have histone acetyltransferase (HAT) activity in vitro. HAT activity maps to the central, most conserved portion of dTAF(II)230 and yTAF(II)130. The HAT activity of dTAF(II)230 resembles that of yeast and human GCN5 in that it is specific for histones H3 and H4 in vitro. Our findings suggest that targeted histone acetylation at specific promoters by TAF(II)250 may be involved in mechanisms by which TFIID gains access to transcriptionally repressed chromatin.

Acetyltransferases↗

PubMed1996

Conversion of L-lactate oxidase to a long chain alpha-hydroxyacid oxidase by site-directed mutagenesis of alanine 95 to glycine.

A mutant form of L-lactate oxidase (LOX) from Aerococcus viridans in which alanine 95 was replaced by glycine was constructed as a mimic of L-lactate monooxygenase but proved instead to be a mimic of the long chain alpha-hydroxyacid oxidase from rat kidney. A95G-LOX keeps oxidase activity with L-lactate at the same level as wild type LOX but has much enhanced oxidase activity with longer chain L-alpha-hydroxyacids, alpha-hydroxy-n-butyric acid, alpha-hydroxy-n-valeric acid, etc., and also the aromatic alpha-hydroxyacid, L-mandelic acid. Kinetic analysis of the activity with these substrates indicates that the reduction of the enzyme bound flavin by substrates is the rate-limiting step in A95G-LOX. The affinity of pyruvate for the reduced enzyme is increased, and sulfite binding to the oxidized enzyme is weaker in A95G-LOX than in native enzyme. Wild type LOX stabilizes both the neutral and anionic flavin semiquinones with a pKa of 6.1, but A95G LOX stabilizes only the anionic semiquinone form. These results strongly suggest that the environment around the N5-C4a region of the flavin isoalloxazine ring is changed by this mutation.

Alanine↗

PubMed1996

Structural similarity between TAFs and the heterotetrameric core of the histone octamer.

A complex of two TFIID TATA box-binding protein-associated factors (TA FIIs) is described at 2.0A resolution. The amino-terminal portions of dTAFII42 and dTAFII62 from Drosophila adopt the canonical histone fold, consisting of two short alpha-helices flanking a long central alpha-helix. Like histones H3 and H4, dTAFII42 and dTAFII62 form an intimate heterodimer by extensive hydrophobic contacts between the paired molecules. In solution and in the crystalline state, the dTAFII42/dTAFII62 complex exists as a heterotetramer, resembling the (H3/H4)2 heterotetrameric core of the histone octamer, suggesting that TFIID contains a histone octamer-like substructure.

Amino Acid Sequence↗