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Biomedical subjects

T Kokubo

Publications and source records attributed to T Kokubo.

At least 19 recordsLinked to original sources
PubMed1995

Molecular cloning of human cDNA for cathepsin K: novel cysteine proteinase predominantly expressed in bone.

We have previously cloned a rabbit cDNA clone (OC-2) from an osteoclast cDNA library by the differential screening. OC-2 was found to encode a novel cysteine proteinase, tentatively called cathepsin K, which is predominantly expressed in osteoclasts. By use of a rabbit OC-2 fragment as a probe, its human counterpart was cloned from a cDNA library of osteoarthritic hip bone. The cloned human cDNA (hOC-2) encoded a protein of 329 amino acid residues and its deduced amino acid sequence showed 94% homology to rabbit cathepsin K. Multiple alignment of amino acid sequences of human cathepsins B, H, L, S and K showed the highest homology of cathepsin K to cathepsin S 48%. Northern blot analysis showed that cathepsin K mRNA is expressed at high levels in some osteoarthritic hip bones and at a very high level in osteoclastoma compared to very low levels in other tissues. These results suggest that cathepsin K is closely involved in human osteoclastic bone resorption.

Amino Acid Sequence↗
PubMed1995

Drug release from a novel self-setting bioactive glass bone cement containing cephalexin and its physicochemical properties.

A novel device containing cephalexin as a model drug using a self-setting bioactive cement based on CaO-SiO2-P2O5 glass was investigated. The device consisted of 95 wt/wt% glass powders and 5 wt/wt% cephalexin powder hardened within 5 min after mixing with a phosphate buffer. After setting, in vitro drug release from homogeneous or heterogeneous drug-loaded cement pellets in a simulated body fluid (SBF) at pH 7.25 and 37 degrees C continued for over 4 weeks. The hardened cement gradually formed low-crystallinity hydroxyapatite with high bioactivity in hard bone tissue and reduced in volume by about 5% during dissolution testing in SBF. Consequently, 30% of the loaded drug was squeezed from the cement system at the initial stage of the drug release, and the remainder released more slowly. Because the heterogeneous system consisting of the cement and drug-loaded pellet avoided the drug-squeezing effect, it showed a longer drug release term than the homogeneous drug-loaded cement. The heterogeneous system using the hardened cement after soaking in SBF at 37 degrees C for 10 days showed very slow drug release at the initial stage because it completely avoided the drug-squeezing effect, and the release was a zero-order pattern.

Cephalexin↗
PubMed1995

Bone-bonding behavior of titanium alloy evaluated mechanically with detaching failure load.

Although titanium (Ti) and Ti alloy are generally classified as bioinert materials in terms of their bonding to bone tissue, it is still unclear whether they bond chemically to bone. In this study, we examined the bone-bonding ability of Ti alloy (Ti-6Al-4V) using smooth-surfaced plates under non-load-bearing conditions. The bone-bonding behavior was evaluated mechanically by means of the detaching test reflecting tensile force. After implantation of the plates into the tibiae of rabbits for 4, 8, 16, and 25 weeks, detaching tests were performed. The failure load of the Ti alloy plates was close to 0 kg at 4 and 8 weeks, but gradually increased with time, reaching 0.334 kg at 16 weeks and 2.852 kg at 25 weeks on average. Histologic examination by Giemsa surface staining and SEM showed no differences between specimens at 8, 16, and 25 weeks, when Ti alloy plate made direct content with bone without any fibrous tissue. By SEM-EPMA, no clear calcium-phosphorus (Ca-P)-rich layer at the interface between the Ti alloy and bone tissue was evident, although a thin bone tissue was observed on the detached Ti alloy plate. The present results indicate that from both mechanical and histologic viewpoints, Ti alloy bonds directly to bone under static conditions after some time, probably more than 8 weeks. The possibility of chemical bone-bonding of Ti alloy was suggested.

Alloys↗
PubMed1995

Apatite coated on organic polymers by biomimetic process: improvement in its adhesion to substrate by glow-discharge treatment.

A dense, uniform, and highly biologically active bone-like apatite layer can be formed in arbitrary thickness on any kind and shape of solid substrate surface by the following biomimetic method at ordinary temperature and pressure: First, a substrate is set in contact with particles of bioactive CaO-SiO2-based glass soaked in a simulated body fluid (SBF) with inorganic ion concentrations nearly equal to those of human blood plasma so that a number of apatite nuclei are formed on the substrate. Second, the substrate is soaked in another solution with ion concentrations 1.5 times those of SBF (1.5SBF) so that the apatite nuclei grow in situ. In the present study, organic polymer substrates were treated with glow-discharge in O2 gas atmosphere, then subjected to the above-mentioned biomimetic process. The induction periods for the apatite nucleation on all the examined organic polymers were reduced from 24 to 6 h, with glow-discharge treatment. The adhesive strengths of the formed apatite layer to the substrates increased from about 4 to 10 MPa for poly(ethylene terephthalate) and poly-ether sulfone, and from 1 approximately 2 to 6 approximately 7 MPa even for poly(methyl methacrylate), polyamide 6 and polyethylene. It is supposed that highly polar groups such as carbonyl, ester, hydroxyl, and carboxyl ones formed by glow-discharge treatment increased the affinity of a silicate ion with the substrates to decrease the induction period, and also increased the affinity of the apatite with the substrate to increase the adhesive strength.(ABSTRACT TRUNCATED AT 250 WORDS)

Adhesiveness↗
PubMed1995

Bioactive bone cement: the effect of amounts of glass powder and histologic changes with time.

A study was conducted to examine the influence of the amount of glass powder added to a bioactive bone cement of our formula on its mechanical and biologic properties. Serial changes in the cement with time were also examined. The bioactive bone cement consisted of CaO-SiO2-P2O5-CaF2 glass powder and bisphenol-a-glycidyl methacrylate resin. Glass powder was added to the cement in 30, 50, 70, and 80% weight ratios. The compressive strengths of the resulting cements (171-239 MPa) were more than double that of polymethylmethacrylate cement (68 MPa). Histologic examination of rat tibiae bearing artificial defects packed with each bioactive cement showed direct bone contact 4 weeks after surgery. The cement with a higher percentage of glass powder showed better direct formation of bone around its periphery with a thicker reactive layer. Under scanning electron microscopic observation, the reactive layer showed increased levels of calcium and phosphorus. Examination of histologic changes up to 26 weeks showed progressive bone formation around the cement and no sign of biodegradation.

Animals↗
PubMed1995

Bone-bonding behavior of three heat-treated silica gels implanted in mature rabbit bone.

Silica gel has been reported to induce apatite nucleation on its surface in vitro and it can act as a stimulant that induces formation of chemical apatite (Ca-P) layers on the surfaces of bioactive glass-ceramics. In this study, apatite formation in response to and the bone-bonding behavior of silica gels implanted in the tibiae of mature rabbits were studied. Implants were made from three silica gels treated at 400, 800, and 1000 degrees C, and the effects of such heat treatment on the above parameters were investigated. The silica gel was made by hydrolysis and polycondensation of tetraethoxysilane in aqueous solution containing polyethylene glycol. Rectangular implants (15 mm x 10 mm x 2 mm) of each heat-treated silica gel were implanted into both tibial bones of mature male rabbits, which were killed 4 or 8 weeks after implantation, and the tibiae containing the implants were dissected out. The bone-implant interfaces were investigated using Giemsa surface staining, contact microradiography, scanning electron microscopy-electron probe microanalysis, and X-ray diffraction. Histologically, no bonding of bone to any of the silica gels was observed at any time postimplantation. Soft tissue was observed at the bone-silica gel interface, but there were no giant foreign body or inflammatory cells. A Ca-P-rich layer was observed only on small areas of the surfaces of the silica gels treated at 400 and 800 degrees C 4 and 8 weeks after implantation. X-ray diffraction analysis confirmed the presence of hydroxyapatite in these Ca-P-rich layers.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1995

Scanning electron microscopy-electron probe microanalysis study of the interface between apatite and wollastonite-containing glass-ceramic and rabbit tibia under load-bearing conditions after long-term implantation.

Glass-ceramic implants containing oxy- and fluoroapatite [Ca10(PO4)6(O, F2)] and beta-wollastonite (CaSiO3) were studied under load-bearing conditions in a segmental replacement model in the tibia of the rabbit. A 16-mm segment of the middle of the tibial shaft was resected at a point distal to the junction of the tibia and the fibula. The defect was replaced by a 15 mm-long hollow, cylindrical implant that was fixed by intramedullary nailing using Kirschner wire. The implants were 9 mm in diameter and 15 mm long bearing a central hole 3.05 mm in dianeter. The rabbits used were killed 6 months, 1 year, 18 months, and 2 years after implantation. The interface between the bone and the glass-ceramic was investigated by scanning electron microscopy-electron-probe microanalysis (SEM-EPMA). None of the glass-ceramic implants broke, and the glass-ceramic had bonded directly to the bone tissue without any intervening soft tissue. A calcium-phosphorus layer (Ca-P layer) was observed at the glass-ceramic/bone interface. This layer was 30-100 microns thick at 6 months after implantation, 60-110 microns thick at 1 year after implantation, 80-200 microns thick at 18 months, and 120-350 microns thick at 2 years. At the lateral surface of the glass-ceramic uncovered by the bone, the calcium-phosphorus layer was 50-80 microns thick at 6 months after implantation, 250-450 microns thick at 1 year, 300 approximately 400 microns thick at 18 months, and 300 microns thick at 2 years. The thickness of the calcium-phosphorus layer increased moderately after long-term implantation. However, it was difficult to estimate the rate of increase in the thickness of calcium-phosphorus layer.

Animals↗
PubMed1994

Interaction between the N-terminal domain of the 230-kDa subunit and the TATA box-binding subunit of TFIID negatively regulates TATA-box binding.

Transcription initiation factor TFIID plays a central role in transcriptional regulation. Drosophila TFIID is a multimeric protein consisting of the TATA box-binding polypeptide (TBP) and a number of tightly associated polypeptides. Previously, the largest subunit of TFIID (p230) was cloned and demonstrated to inhibit the TATA-box binding of TBP in the absence of other subunits. Here we demonstrate that p230 contains at least two sites of interaction with TBP and that the N-terminal site mediates both strong physical interactions with TBP and inhibition of the TBP function. A detailed mutagenesis study shows that the inhibitory domain is indistinguishable from the strong TBP-binding domain, thus indicating that interaction of the p230 N-terminal region with TBP may directly control TATA-box binding.

Amino Acid Sequence↗
PubMed1994

Molecular cloning of Drosophila TFIID subunits.

Transcription initiation factor TFIID is a multisubunit complex containing a TATA-box-binding factor (TFIID tau/TBP) and associated polypeptide factors (TAFs) with sizes ranging from M(r) approximately 20,000 to > 200,000. As a result of direct promoter interactions, TFIID nucleates the assembly of RNA polymerase II and other initiation factors into a functional preinitiation complex. Although the native TFIID complex mediates both basal and activator-dependent transcription in reconstituted systems, TBP itself is competent for only basal transcription. Thus, TAFs are essential cofactors for regulated transcription. The complementary DNAs encoding the p230 (M(r) 230,000), p110 and p85 subunits of TFIID have recently been cloned. Here we report the molecular cloning and characterization of the p62, p42, p28 and p22 subunits. These participate in a network of heterogeneous protein-protein interactions within TFIID. Sequence similarities between p62/p42 and the histones H4/H3, respectively, suggest that these subunits have a functional relationship with chromatin.

Amino Acid Sequence↗
PubMed1994

The role of hydrated silica, titania, and alumina in inducing apatite on implants.

Pure soluble silica prepared by a sol-gel method induced bone-like hydroxyapatite formation onto its surface when the silica was immersed in a simulated body fluid (SBF), whereas silica glass and quartz did not. This finding directly supports the hypothesis that hydrated silica plays an important role in biologically active hydroxyapatite formation on the surfaces of bioactive glasses and glass-ceramics, which leads to bone-bonding. Gel-derived titania is also a hydroxyapatite inducer because of its abundant TiOH groups. These results provide further insight into the unique osseointegration of titanium and its alloys. It is suspected that gel-derived titania develops an apatite layer by taking calcium and phosphate from the body fluid, thus producing bone-bonding. Although sufficient AlOH groups may remain in the alumina gel, they do not serve to initiate apatite generation when immersed in SBF. This phenomenon explains the fact that an intermediate fibrous tissue is usually found to separate the alumina implant from bone. One may infer that both abundant OH groups and negatively charged surfaces of gel-derived silica and titania are important for hydroxyapatite induction. material which possesses and/or develops both a negatively charged surface and abundant OH groups in a physiologically-related fluid is most likely to be an efficient apatite inducer. Such materials are suitable candidates to serve as bone-bonding biomaterials.

Aluminum Oxide↗
PubMed1994

Ultrastructural study of the A-W GC-bone interface after long-term implantation in rat and human bone.

The interface between apatite- and wollastonite-containing glass-ceramic (A-W GC) and bone after long-term implantation was studied by scanning and transmission electron microscopy (SEM and TEM) using rat and human specimens. First, particles of A-W GC (100-220 microns in diameter) were implanted into rat tibiae, and specimens were prepared for observation at 24, 48, 72, and 96 weeks after the operation. These long-term specimens showed an A-W GC-bone interface different from that at an earlier stage, which was investigated in our previous studies. SEM showed that the Ca-P-rich layer was wider, suggesting that leaching of ions from the A-W GC had continued even after bonding with bone. In some regions, the material particles were evidently replaced by the bone. TEM showed that the intervening apatite layer had become indistinct, and that A-W GC had intermingled with bone at the interface. In some regions, the surface of the A-W GC was degraded. These findings suggest that the surface region of A-W GC is slowly replaced by bone. Second, a human bone specimen, which included A-W GC particles (300-700 microns in diameter) implanted as a bone filler for about 75 weeks was harvested and investigated. Excellent A-W GC-bone bonding was observed, and the ultrastructure of the interface was similar to that in rats after long-term implantation. This finding demonstrated that A-W GC possibly worked in human bone in the same way as in rat bone, showing excellent bioactivity.

Adult↗
PubMed1994

Osteoclastic resorption of apatite formed on apatite- and wollastonite-containing glass-ceramic by a simulated body fluid.

We immersed mirror-polished apatite- and wollastonite-containing glass-ceramic (A-W GC) disks in a simulated body fluid (SBF) for 5 days to form bonelike apatite on their surface. Neonatal rabbit bone cells were cultured on these or on plain A-W GC disks for 10, 24, and 48 h. We observed the substrates by scanning electron microscopy after treating them with pronase E plus EDTA to remove all cells except osteoclasts. Osteoclasts with a non-motile appearance formed no lacunae on the plain A-W GC, whereas on the bonelike apatite formed on A-W GC by the SBF, actively moving osteoclasts made many tracklike resorption lacunae. These were evident even after 10 h of culture and became more extensive after longer culture periods. The bonelike apatite was therefore a more suitable medium than plain A-W GC for maintaining osteoclast activity. This study demonstrated in vitro osteoclastic resorption of bonelike apatite formed on A-W GC by an SBF. It suggests that the apatite layer, through which a surface-active ceramic bonds to bone in vivo, can be resorbed by osteoclasts and subjected to bone remodeling.

Animals↗
PubMed1994

Bioactive glass-ceramic containing crystalline apatite and wollastonite initiates biomineralization in bone cell cultures.

Rat bone cells were cultured in the presence of bioactive glass-ceramic containing crystalline apatite and wollastonite. Scanning electron microscopy observations of the surface of the seeded ceramic disks revealed that cells attached, spread, and proliferated on the material surface. Soaking in cell-free culture medium showed that no change occurred in the surface structure. However, when cultured with bone cells and observed under a transmission electron microscope, an electron-dense layer was noted initially at the surface of the material, before bone formation occurred. In addition, energy-dispersive X-ray microanalysis demonstrated the presence of calcium and phosphorus in this layer. Progressively, during the following days of culture, active osteoblasts synthetized and laid down an osteoid matrix composed of numerous collagen fibrils arranged either parallel or perpendicularly to the first-formed electron-dense layer. Mineralization initiated on the ceramic surface dispersed then along the collagenous fibrils, leading to a mineralized matrix which surrounded the ceramic particles. These results demonstrate the capacity of apatite-wollastonite glass ceramic to initiate biomineralization in osteoblast cultures and to achieve a direct bond between the surface apatite layer of the bioactive glass-ceramic and the mineralized bone matrix.

Animals↗
PubMed1994

Interaction between drugs and pressure-sensitive adhesives in transdermal therapeutic systems.

Release experiments with four drugs using representative pressure-sensitive adhesive (PSA) matrices were performed at 37 degrees C, and drug-PSA polymer interaction was determined by the Williams, Landel, and Ferry (WLF) equation. Two acrylic-type [2-ethylhexylacrylate and acrylic acid copolymer (2EHA/AA) or acrylamide copolymer (2EHA/AAm)], one rubber-type (a mixture of high and low molecular weight polyisobutylene), and one silicone-type PSA were used, and dipropylphthalate (PP), aminopyrine (AMP), ketoprofen (KP), and lidocaine (LC) were selected as model drugs because of their molecular size and functional groups. PSA containing acrylic acid (2EHA/AA) strongly interacted with the amide LC, with the tertiary amine AMP, and with the carboxylic acid KP; PSA-containing acrylamide (2EHA/AAm), however, did not interact with LC or AMP, although it markedly interacted with KP. The rubber-type and silicone-type PSAs, composed of no or only a few polar functional groups, did not interact with any of the drugs used in this experiment. Therefore, the diffusion coefficient of the drugs through PSA was influenced by the drug-PSA polymer interaction, and the extent of this interaction can be estimated by the relationship between the drug concentrations in the PSA and their diffusion coefficients.

Acrylates↗
PubMed1994

Successful treatment of cytophagic histiocytic panniculitis with modified CHOP-E. Cyclophosphamide, adriamycin, vincristine, predonisone, and etoposide.

A 27-year-old woman developed generalized subcutaneous painful nodules, fever, abnormal liver function, a bleeding tendency, and pancytopenia. Skin biopsies revealed the lobular panniculitis with a morphologically benign histiocytic infiltration with prominent phagocytosis. Leukophagocytosis and erythrophagocytosis were also present in the bone marrow. The diagnosis of cytophagic histiocytic panniculitis was made. The patient received polychemotherapy with cyclophosphamide, Adriamycin, and vincristine on day 1, prednisone on day 1-5 (modified CHOP), with the addition of etoposide (E). This regimen was repeated 8 times every 3 weeks. The patient obtained a complete clinical remission that has lasted almost 2 years after the completion of chemotherapy. Thus we suggest modified CHOP-E chemotherapy for an effective treatment for the aggressive form of cytophagic histiocytic panniculitis.

Adult↗
PubMed1994

A case of Recklinghausen neurofibromatosis associated with membranous nephropathy.

A 68-year-old woman who had a history of Recklinghausen neurofibromatosis from 1964 showed nephrotic syndrome in 1989. Renal biopsy revealed membranous nephropathy. Various suspected causes of secondary membranous nephropathy were not found. Coexistence of Recklinghausen neurofibromatosis and membranous nephropathy has rarely been reported.

Aged↗