Biomedical subjects

Y Kadoya

Publications and source records attributed to Y Kadoya.

At least 37 records · Page 2Linked to original sources

PubMed1999

Significant role of laminin-1 in branching morphogenesis of mouse salivary epithelium cultured in basement membrane matrix.

Mouse submandibular epithelium shows branching morphogenesis in mesenchyme-free conditions when covered with a basement membrane matrix (Matrigel) in medium supplemented with epidermal growth factor. In the present study, the role of laminin-1 (LN1), a major glycoprotein of Matrigel, in this culture system was defined. When the epithelium was cultured in a LN1-nidogen gel, the epithelium showed much branching, comparable to that observed with Matrigel. By electron microscopy, only a felt-like matrix was formed on the epithelial surface in the LN1-nidogen gel cultures, while an organized basal lamina structure was formed on the epithelial surface in direct or transfilter recombination cultures with mesenchyme. Next, the epithelium covered with Matrigel was cultured in medium containing either biologically active peptides from LN1, IKVAV-including peptide (2097-2108), AG10 (2183-2194), AG32 (2370-2381) or AG73 (2719-2730) from the alpha1 chain, or YIGSR-including peptide (926-933) from the beta1 chain. Only AG73 (RKRLQVQLSIRT from the alpha1 chain carboxyl-terminal globular domain) inhibited the epithelial branching in Matrigel. These results suggest that LN1-nidogen can support the branching morphogenesis of submandibular epithelium even if LN1-nidogen is not assembled into an intact basal lamina, and that the AG73 sequence is an important site on LN1, which interacts with submandibular epithelial cells.

Amino Acid Sequence↗

PubMed1999

Anteroposterior and rotational movement of femur during knee flexion.

This study was designed to analyze anteroposterior and rotational movement of femoral condyles during knee flexion from 15 degrees to 90 degrees using magnetic resonance imaging. After a pilot study, scans were made in 10 healthy male Japanese volunteers. When centers of the circular profiles of posterior femoral condyles were used as reference points, the medial and lateral femoral condyles displaced posteriorly 1.9 +/- 0.8 mm and 2.3 +/- 0.5 mm, respectively (mean +/- standard error). Duplicate examinations on two separate occasions revealed the accuracy of this procedure was in the range of 1 to 2 mm. These results have confirmed that femoral rollback occurs in the unloaded normal knee during flexion from 15 degrees to 90 degrees, but its magnitude is small (2 mm). The results obtained in this in vivo measurement of anteroposterior movement of the femoral condyles have relevance for total knee replacement design.

Adult↗

PubMed1998

Measurements of zeta potentials of particulate biomaterials in protein-rich hyaluronan solution with changes in pH and protein constituents.

This study was undertaken to determine the zeta potentials of particulate biomaterials in three types of protein-rich hyaluronan solution with changes in pH; a microelectrophoretic method was used. For the purpose of determining the pH value of synovial fluid in various inflammatory conditions, we collected synovial fluid samples from joints with osteoarthritis (OA), rheumatoid arthritis (RA), and those undergoing revisions arthroplasties. The mean values of the pH in the synovial fluid from joints with OA, RA, and revision arthroplasty were shown to be 7.9, 7.5, and 8.1, respectively. The pH-zeta potential curves obtained differed, depending on the biomaterial and the medium. Addition of gamma-globulin to the medium reduced the absolute value of the zeta potentials of some of the biomaterials. The findings of this study suggest that the electrophoretic behaviors of the particulate biomaterials tested in this study are affected by the protein constituents of and pH changes in protein-rich synovial fluid. The values we obtained will be useful as reference standards and will also aid in the study of the surface phenomena of biomaterials.

Biocompatible Materials↗

PubMed1998

Laminin alpha1 chain G domain peptide, RKRLQVQLSIRT, inhibits epithelial branching morphogenesis of cultured embryonic mouse submandibular gland.

Active sequences from the laminin alpha1 and alpha2 chain carboxyl-terminal globular domains (G domain) have been identified by screening overlapping synthetic peptides in a number of biological assays (Nomizu et al. [1995] J. Biol. Chem. 270:20583-20590; Nomizu et al. [1996] FEBS Lett. 396:37-42). We have tested the activity of these peptides in submandibular gland explants of embryonic day 13 mice to determine the functional sites involved in organ development. The laminin alpha1 chain peptide, RKRLQVQLSIRT (residues 2719-2730 and designated AG-73), significantly inhibited epithelial branching morphogenesis. In contrast, other cell adhesive laminin alpha1 chain peptides including the AASIKVAVSADR and NRWHSIYITRFG failed to inhibit the branching. MG-73, a homologue of AG-73 from the laminin alpha2 chain, did not inhibit the branching. The alpha2 chain peptide had no effect, which may be due to the low levels of this laminin chain in day 13 mice. Laminin alpha2 chain-specific monoclonal antibodies strongly reacted with the basement membranes of developed acini but only weakly stained embryonic day 13 submandibular epithelium. The expression of E-cadherin and alpha6 integrin, as detected by immunofluorescence, were unchanged in both AG-73 and control scramble peptide-treated epithelial cells of the explants. In contrast, immunostaining of nidogen/entactin showed that explants treated with AG-73 for 3 days had a discontinuous basement membrane. Explants treated for 3 days with control peptide showed a normal basement membrane. These results suggest that the region containing the AG-73 sequence of the laminin alpha1 chain is crucial for development of submandibular gland at early embryonic stages. The discontinuous basement membrane in AG-73-treated explants may indicate an important role for this region in basement membrane assembly.

Animals↗

PubMed1998

Wear and osteolysis in total joint replacements.

UNLABELLED: This presentation summarizes the results of our recent studies on the pathogenesis of osteolysis around total joint arthroplasties. First, interface tissues with adjacent bone were retrieved and histopathologically investigated with reference to the cells on the bone surface. Secondly, polyethylene particles were extracted with the tissue digestion method and characterized with scanning electron microscopy. Finally, an animal model for osteolysis was created and various interface conditions were compared concerning their resistance to particle migration. Histopathological examinations demonstrated that active bone formation, regarded as a repair process, was the commonest feature, even in revised cases. They also highlighted the role played by macrophages, not as cells producing inflammatory mediators which could activate osteoclasts, but as cells primarily responsible for the bone loss in osteolytic lesions. Among the particle species present, only polyethylene particles were shown to play a significant role in macrophage recruitment and subsequent osteolysis. A quantitative extraction of polyethylene particles showed a significant difference in the "number" of particles between osteolysis positive and negative cases whereas the "sizes" of particles were similar in these two groups. The critical number of particles for osteolysis was around 1 x 10(10) particles/g tissue and the cellular reaction against phagocytosable particles accumulated over this concentration may be the prerequisite for progression of osteolysis. The animal model for osteolysis indicated that the progression of osteolysis depends on the integrity of the bone-implant interface. We suggest that the solid fixation of the prosthesis performed by current techniques (e.g., improved cementing technique, hydroxyapatite coating) is beneficial for preventing particle migration and subsequent osteolysis. CLINICAL RELEVANCE: Osteolysis induced by particulate wear debris from implant materials has been recognized as the major cause of long-term failure in total joint replacements. However, the development of preventive measures for this phenomenon has not been successful because the mechanism in which wear particles cause osteolysis is not quite clear. On the basis of results obtained in this study, we believe that the basic strategy for addressing the problem of osteolysis is to reduce the "number" of accumulated wear particles in the interface tissues. This could be achieved either by improving the materials or the geometry of the articulating counterface. Another possibility is to increase the integrity of the bone-implant interface to prevent particle migration. It is important to note that pre-clinical testing of materials and prosthetic designs should include an analysis of the characteristics of the particle generated (e.g., size and number). The widespread bone formation, even in revised cases, is encouraging in view of "conservative treatment" of aseptic loosening. Assuming that bone loss in aseptic loosening is not a remorseless process, some form of intervention, whether mechanical or pharmacological, might be possible to tip the balance more in favour of bone formation than resorption. A comprehensive understanding of the bone reactions in osteolysis, including the basic mechanisms of bone loss, shown in this study, are decisive for the development of preventive measures that may minimize the clinical impact of this phenomenon.

Animals↗

PubMed1998

The EGF system in fetal development.

Immunohistochemical localization of the epidermal growth factor receptor (EGFR) disclosed a wide distribution of this protein in diverse fetal tissues. This review focuses on some of the vast work on the expression and operation of the EGFR and its principal ligands in fetal organogenesis.

Animals↗

PubMed1997

Substructures of the acinar basement membrane of rat submandibular gland as shown by alcian blue staining and cryo-fixation followed by freeze-substitution.

The ultrastructure of the epithelial basement membrane was studied in the acinar cells of adult rat submandibular glands after: (i) immersion fixation in 0.1 M sodium cacodylate buffer (CB, pH 7.2) containing 2.5% glutaraldehyde (GA) and alcian blue (0.5%) in conjunction with microwave irradiation, (ii) perfusion fixation with 2.5% GA in CB, (iii) rapid freezing followed by freeze-substitution (RF-FS) with 1% GA in acetone, and (iv) RF-FS with 2% OsO4 in acetone. The specimens were post-fixed with 1% OsO4 in CB after methods (i) and (ii) but not (iii) and (iv). Fixed specimens were embedded in epoxy resin and the ultrathin sections were cut, stained with both lead and uranium, and observed under a transmission electron microscope. Various substructural components could be seen in the acinar basement membrane. In the specimens processed by method (iv), a clear meshwork structure could be found just beneath the basal plasma membrane. This meshwork could not be seen in the specimens processed by method (iii) but thin filaments of approximately 100 nm in length extending from the epithelial base toward the connective tissue space were evident. By methods (i) and (ii), an electron dense 30-75 nm layer could be seen subjacent to basal cell membranes. By method (ii), particularly, thick threads connecting this layer to collagen fibres in the connective tissue were stained with alcian blue. Lamina lucida was not identified.

Alcian Blue↗

PubMed1997

Wear particulate species and bone loss in failed total joint arthroplasties.

The aim of this study was to evaluate the relative contribution of polyethylene, metal, and polymethylmethacrylate (cement) particles to the overall bone loss in aseptic loosening. Twenty-four interface tissues with adjacent bone were obtained during 17 revision total joint arthroplasties (11 hips and six knees). Osteoclasts and macrophages were identified immunohistochemically on the bone surface. The length of the bone surface in contact with these cell types was measured and analyzed with reference to the particulate species present within the fibrous interface. The presence of abundant polyethylene particles significantly increased the proportion of the bone surface in contact with macrophages but did not have a significant influence on that of osteoclasts. Osteoclastic bone resorption was significantly more extensive in the presence of metal particles. In contrast, the presence of cement particles did not have a significant influence on macrophage or osteoclast coverage of the bone surface. These results highlight the significance of polyethylene particles in macrophage recruitment and subsequent osteolysis and suggest a different mechanism of bone loss related to metal, namely mediation through osteoclastic activities. The relative contribution of cement particles was negligible and needs reevaluation in light of evidence provided by others.

Adult↗

PubMed1997

Expression of epidermal growth factor receptor in fetal mouse submandibular gland detected by a biotinyltyramide-based catalyzed signal amplification method.

Branching morphogenesis of the fetal mouse submandibular gland (SMG) can be modulated in vitro by stimulation or inhibition of the epidermal growth factor receptor (EGFR). Because the mRNAs for EGF and EGFR are detectable in RNA of SMG rudiments isolated directly from fetuses, the EGF system probably operates physiologically as a regulator of SMG morphogenesis. However, neither EGFR protein nor its precise cellular localization has been characterized in the fetal SMG. Here we show EGFR protein in fetal mouse SMG by immunoprecipitation, affinity labeling, ligand-induced autophosphorylation, and immunohistochemistry. SMGs from E16 fetuses (day of vaginal plug = E0) were labeled with [35S]-cysteine/methionine and homogenized. After addition of specific antibody to EGFR, the immunoprecipitate was isolated, resolved by polyacrylamide gel electrophoresis, and detected by autoradiography. A single band of 170 kD was detected, corresponding to the EGFR protein. Affinity labeling with [125I]-EGF of the membrane fraction of E18 SMG also revealed a prominent band at 170 kD, showing that this EGFR protein can bind specifically to its ligand. Incubation of SMG membranes from E18 fetuses with EGF in the presence of [gamma-32P]-ATP, followed by immunoprecipitation with anti-phosphotyrosine antibody also showed a single band at 170 kD, demonstrating autophosphorylation of the EGFR in response to binding of its ligand. Immunohistochemical localization of the cellular sites of EGFR in the fetal SMG required use of a catalyzed signal amplification procedure, with biotinyltyramide as the amplifying agent. EGFR was localized predominantly, if not exclusively, in cell membranes of epithelial cells of the rudiment, whereas staining of mesenchymal cells was equivocal. Staining was strongest on duct cells, and weak on cells of the end-pieces. These findings clearly show that a functional EGFR protein is expressed in fetal SMG chiefly, if not exclusively, on epithelial cells.

Affinity Labels↗

PubMed1997

Importance of nidogen binding to laminin gamma1 for branching epithelial morphogenesis of the submandibular gland.

Epithelial-mesenchymal interactions are major driving forces for the development of most solid organs. The importance of these interactions was first shown for the embryonic submandibular gland more than 40 years ago. We here present evidence that interactions between two basement membrane components, nidogen (entactin) and laminin gamma1 chain, could be important for epithelial-mesenchymal interactions in this gland. Nidogen mRNA was detected by in situ hybridization in the mesenchyme, and yet the protein was detected in epithelial and endothelial basement membranes. The role of nidogen-laminin interactions for epithelial morphogenesis was studied by applying antibodies to submandibular gland organ cultures. Antibodies reacting strongly with the nidogen-binding site of laminin gamma1 chain drastically perturbed branching epithelial morphogenesis. Electron microscopy of the epithelial-mesenchymal interface showed that blocking antibodies disrupted the formation of the basement membrane. Epidermal growth factor was shown to increase the expression of nidogen in mesenchyme, and could counteract the effect of the blocking antibodies. We suggest that nidogen could be an important mesenchymal factor for submandibular gland development.

Animals↗

PubMed1997

The size and shape of particulate polyethylene wear debris in total joint replacements.

Osteolysis induced by wear particles has been recognized as one of the major causes of long-term failure in total joint replacements. However, little is currently known about the exact nature of particles, as the particles are too small to be characterized by light microscopy. In this study, ultra-high molecular weight polyethylene (UHMWPE) particles retrieved from ten cases (six cemented and four uncemented) for Freeman type conforming tibiofemoral total knee replacements (TKRs), three Charnley total hip replacements (THRs) and five Imperial College/London Hospital double cup surface hip replacements for aseptic loosening were extracted using a high-performance method with ultracentrifugation and characterized by scanning electron microscopy. The equivalent circle diameter (ECD) of all 18 cases ranged from 0.40 to 1.15 microns (Mean +/- SE = 0.70 +/- 0.05 micron, median = 0.67 micron). The aspect ratio was 1.50 to 2.04 (Mean +/- SE = 1.75 +/- 0.04, median = 1.73), and roundness was 1.24 to 2.34 (Mean +/- SE = 1.61 +/- 0.07, median = 1.65). The numbers of particles were 5.2 x 10(8) to 9.17 x 10(10)/g tissue (Mean +/- SE = 1.42 x 10(10) +/- 5.41 x 10(9)/g tissue, median = 7.04 x 10(9)). The number of polyethylene (PE) particles/g tissue in TKRs was significantly larger than that in THRs (1.04 x 10(10)/g tissue and 2.16 x 10(9)/g tissue respectively, median, p = 0.03, Mann-Whitney U test). Unstable fixation of the tibial PE component might account for the accumulation of a large number of PE particles in the interface tissue.

Biocompatible Materials↗

PubMed1997

Number of polyethylene particles and osteolysis in total joint replacements. A quantitative study using a tissue-digestion method.

Our aim was to analyse the influence of the size, shape and number of particles on the pathogenesis of osteolysis. We obtained peri-implant tissues from 18 patients having revision surgery for aseptically loosened Freeman total knee replacements (10), Charnley total hip replacements (3) and Imperial College/London Hospital double-cup surface hip replacements (5). The size and shape of the polyethylene particles were characterised using SEM and their concentration was calculated. The results were analysed with reference to the presence of radiological osteolysis. The concentration of polyethylene particles in 6 areas with osteolysis was significantly higher than that in 12 areas without osteolysis. There were no significant differences between the size and shape of the particles in these two groups. We conclude that the most critical factor in the pathogenesis of osteolysis is the concentration of polyethylene particles accumulated in the tissue.

Chromatography, High Pressure Liquid↗

PubMed1997

Nitric oxide synthase is expressed in human macrophages during foreign body inflammation.

Although nitric oxide (NO) is a well documented effector molecule in rodent macrophages, its significance in human mononuclear phagocytic cells has been controversial. The foreign body inflammatory reaction around loosened joint replacement implants leads to formation of an osteolytic granulomatous pseudo-synovial membrane rich in activated macrophages. We studied 13 specimens of interface membrane tissue collected from revision surgery of aseptically loosened hip and knee prostheses for the presence of inducible NO synthase (iNOS). The presence of iNOS was demonstrated immunohistochemically in 10 of these specimens. Within the tissue this enzyme was confined to macrophages and vascular endothelial cells. iNOS activity was demonstrated biochemically by measuring the calcium-independent generation of citrulline from L-arginine, and the presence of iNOS mRNA was demonstrated using reverse transcriptase polymerase chain reaction. NO synthesis in the interface tissue may be an important factor in the maintenance of the inflammatory and osteolytic processes.

Adult↗

PubMed1996

Bone formation and bone resorption in failed total joint arthroplasties: histomorphometric analysis with histochemical and immunohistochemical technique.

To elucidate the reactions of bone around aseptically loosened total joint arthroplasties, 24 interface tissues with adjacent bone were obtained in 17 revision operations (11 hips and six knees). The morphology of the bone surface next to the interface membrane was investigated with histochemical and immunohistochemical techniques and then histomorphometrically analysed. One-third of the total bone surface. 32.69 +/- 5.16% (mean +/- SE) (n = 24), showed positive alkaline phosphatase activity. The bone surface in contact with the cells positive for CD11b (a macrophage marker) amounted to 19.33 +/- 5.16% (n = 24). The proportion of the osteoclastic bone resorption estimated by vitronectin receptor expression was 7.67 +/- 1.82% (n = 21). Tissues retrieved from the sites where radiographic evidence of osteolysis was present (n = 12) had a significantly larger extent of the bone surface in contact with CD11b-positive cells than did the tissues from areas without osteolysis (n = 12, p = 0.0067, Mann-Whitney U test), whereas no significant difference was observed in the extent of osteoclastic bone resorption. These data demonstrate that active bone formation, regarded as a repair process, is the most common feature even in revised cases. They also highlight the role played by macrophages, not as cells producing inflammatory mediators that could activate osteoclasts, but as cells primarily responsible for the bone loss in osteolytic lesions.

Adult↗

PubMed1996

Assessment of the role of GM-CSF in the cellular transformation and the development of erosive lesions around orthopaedic implants.

Granulocyte-macrophage colony stimulating factor (GM-CSF) is a hematopoietic growth factor with a regulatory effect on the transformation of immature macrophages into multinucleated giant cells (MNGC) that exhibit phenotypic and functional characteristics of osteoclasts (OC). The authors analyzed the bone implant interface membranes harvested from 15 patients with failed total joint replacements for the production and tissue distribution of GM-CSF and interleukin-1 (IL-1). Immunohistology and liquid culture were employed to assess the contribution of these factors in the recruitment of macrophages and the development of bone resorbing MNGC at these sites. This process has been implicated in osteoclastic bone resorption, bone, and bone marrow necrosis adjacent to orthopaedic implants. Histologic assessment of the interface indicated the presence of granuloma and a variable number of MNGC in 11 cases. Four cases showed sites of intramembranous formation of osteoid and mineralized bone that was accompanied by normal bone marrow in two cases. Granulocyte-macrophage colony stimulating factor was expressed by a distinct subset of phagocytic macrophages in the lining layer on the implant side. interleukin-1-positive cells outnumbered those stained for GM-CSF. Stimulation of cultured cells with prosthetic metal particulate material showed marked similarity in the expression of these cytokines to cultures treated with lipopolysaccharide (LPS) or phytohemagglutinin (PHA). The induction of GM-CSF production in the lining layer where small MNGC develop indicates that these cells differentiate locally following the phagocytosis of particulate wear debris. In conclusion, GM-CSF promotes the proliferation and early stages of fusion and development of MNGC responsible for osteolysis at these sites. These results also highlight the capacity of the interface to display both osteogenic and inflammatory characteristics. Collectively, the findings suggest that the local bone marrow could participate in the development of the interface as a source of myeloid cells in addition to the capacity of marrow stroma to generate various osteogenic cells essential for the ingrowth of bone into prosthetic implants.

Adult↗

PubMed1995

Antibodies against domain E3 of laminin-1 and integrin alpha 6 subunit perturb branching epithelial morphogenesis of submandibular gland, but by different modes.

Branching epithelial morphogenesis requires interactions between the surrounding mesenchyme and the epithelium, as well as interactions between basement membrane components and the epithelium. Embryonic submandibular gland was used to study the roles of two mesenchymal proteins, epimorphin and tenascin-C, as well as the epithelial protein laminin-1 and one of its integrin receptors on branching morphogenesis. Laminin-1 is a heterotrimer composed of an alpha 1 chain and two smaller chains (beta 1 and gamma 1). Immunofluorescence revealed a transient expression of laminin alpha 1 chain in the epithelial basement membrane during early stages of branching morphogenesis. Other laminin-1 chains and alpha 6, beta 1, and beta 4 integrin subunits seemed to be expressed constitutively. Expression of epimorphin, but not tenascin-C, was seen in the mesenchyme during early developmental stages, but a mAb against epimorphin did not perturb branching morphogenesis of this early epithelium. In contrast, inhibition of branching morphogenesis was seen with a mAb against the carboxy terminus of laminin alpha 1 chain, the E3 domain. An inhibition of branching was also seen with a mAb against the integrin alpha 6 subunit. The antibodies against laminin alpha 1 chain and integrin alpha 6 subunit perturbed development in distinct fashions. Whereas treatment with the anti-E3 resulted in discontinuities of the basement membrane at the tips of the branching epithelium, treatment with the mAb against alpha 6 integrin subunit seemed to leave the basement membrane intact. We suggest that the laminin E3 domain is involved in basement membrane formation, whereas alpha 6 beta 1 integrin binding to laminin-1 may elicit differentiation signals to the epithelial cells.

Animals↗

PubMed1995

Neovascularisation and the induction of cell adhesion molecules in response to degradation products from orthopaedic implants.

OBJECTIVES: To characterise the cellular interactions and the mechanisms involved in the recruitment of inflammatory macrophages and T cells to the bone implant interface in 30 patients with aseptically loosened orthopaedic prostheses. METHODS: Cell adhesion molecules E-selectin, VCAM-1, ICAM-1 and the receptors LFA-1 and CR3 were immunolocalised on cryostat sections of the bone-implant interface obtained during revision arthroplasty. The percentage of expression on vascular endothelium was determined on serial sections. RESULTS: E-selectin was upregulated on different vessels in 21 patients. Its expression correlated strongly with the presence of metal wear debris. VCAM-1 was detected on vessels in six patients only, and was coexpressed with E-selectin in three patients with metal debris. VCAM-1 was more frequently observed in the lining cells on the implant side. ICAM-1 was upregulated on vessels on the bone side only in 13 patients, and was more strongly expressed on aggregates of macrophages and multinucleated giant cells on the implant side. These macrophage aggregates coexpressed both ICAM-1 and CR3. CONCLUSION: Our findings implicate the contribution of three different pathways in the recruitment of inflammatory cells to the joint in response to orthopaedic implant wear particles. The association of E-selectin expression and metal debris may suggest hypersensitivity reactions. Finally, the simultaneous expression of ICAM-1 and CR3 on the same macrophages on the implant side may predict an additional function of these molecules in homotypic adhesion/cell aggregation that precede differentiation of phagocytes to multinucleated giant cells.

Adult↗

PubMed1995

A new phenylpropanoid glucoside and other constituents of Oenanthe javanica.

Three glucosides (1-3), together with known phenylpropanoids and polyacetylenes were isolated from Oenanthe javanica Blume (DC.). Glucoside 1, named oenanthoside A, is a new phenylpropanoid glucoside and was determined to be 2,3-methylenedioxy-5-allylphenyl beta-D-glucopyranoside, mainly by NMR techniques. Glucosides 2 and 3 were identified as eugenyl beta-D-glucopyranoside and pinoresinol beta-D-glucopyranoside, respectively. These were found for the first time in this plant.

Anti-Bacterial Agents↗