Authorship：Last author,　Corresponding author   Publishing type：Research paper (scientific journal)  

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Language：Japanese   Publisher：日本バイオマテリアル学会  

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Language：Japanese   Publisher：(一社)日本人工臓器学会  

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Language：Japanese   Publisher：(一社)日本人工臓器学会  

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Authorship：Corresponding author   Publishing type：Research paper (scientific journal)   Publisher：MDPI AG  

Cell aggregates have been applied in various fields such as regenerative medicine and drug toxicity testing. We have shown that H-(Lys-Pro)12-OH (KP24), a repeating sequence of lysine (Lys) and proline (Pro), induces uniformly sized cell aggregates simply by its presence in cell suspension. In this study, we considered that this peptide could be applied to a three-dimensional culture substrate that can induce uniform cell aggregates by immobilizing it on the substrate. Therefore, mouse fibroblasts (L929) were seeded on KP24-immobilized glass substrates and cell behavior was observed. We also seeded human-derived cells, namely, human mesenchymal stem cells (hMSC), on KP24-immobilized substrates and characterized their cell assemblies. Furthermore, KP24 was chemically immobilized on the substrate surface, which allowed us to trace the mechanism of KP24–cell interaction. As a mechanism analysis of the cell aggregation ability of KP24, we investigated whether KP24 interacts with the cell surface without being incorporated into the cell.

DOI： 10.3390/pr10091779

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Authorship：Corresponding author   Publishing type：Research paper (scientific journal)  

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Authorship：Corresponding author   Publishing type：Research paper (scientific journal)  

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Language：English   Publishing type：Research paper (scientific journal)  

Decellularized tissue is expected to be utilized as a regenerative scaffold. However, the migration of host cells into the central region of the decellularized tissues is minimal because the tissues are mainly formed with dense collagen and elastin fibers. This results in insufficient tissue regeneration. Herein, it is demonstrated that host cell migration can be accelerated by using decellularized tissue with a patterned pore structure. Patterned pores with inner diameters of 24.5 ± 0.4 μm were fabricated at 100, 250, and 500 μm intervals in the decellularized vascular grafts via laser ablation. The grafts were transplanted into rat subcutaneous tissue for 1, 2, and 4 weeks. All the microporous grafts underwent faster recellularization with macrophages and fibroblast cells than the non-porous control tissue. In the case of non-porous tissue, the cells infiltrated approximately 50% of the area four weeks after transplantation. However, almost the entire area was occupied by the cells after two weeks when the micropores were aligned at a distance of less than 250 μm. These results suggest that host cell infiltration depends on the micropore interval, and a distance shorter than 250 μm can accelerate cell migration into decellularized tissues.

DOI： 10.1039/d1tb02271g

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：Elsevier BV  

DOI： 10.1016/j.jconrel.2020.07.041

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：Elsevier BV  

DOI： 10.1016/j.polymdegradstab.2020.109240

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Acellular blood vessels possess high potential to be used as tissue-engineered vascular scaffolds. Previously, a high patency was achieved for an Arg-Glu-Asp-Val (REDV) peptide-immobilized small-diameter acellular graft in a minipig model. Results revealed the potential of the peptide to capture a circulating cell and also to suppress fibrin clot deposition. Here, the effect of REDV peptide density on the blood response under ex vivo blood perfusion conditions was investigated. When endothelial cells or platelets were seeded under static conditions, the number of adherent endothelial cells increased with the increase in peptide density. Platelets scarcely adhered on the surface where the peptide density was above 18.9 × 10-4 molecules per nm3. Fibrin clot deposition and circulating cell capture were evaluated in a minipig extracorporeal circulatory system. The fibrin clot did not form on the peptide-immobilized surface, in the range of peptide modification density that was evaluated, whereas the unmodified surface was covered with microthrombi. REDV-specific blood circulating cells were captured on the peptide-immobilized surface with a density above 18.9 × 10-4 molecules per nm3. These results illustrated, under ex vivo blood perfusion conditions, that the REDV-immobilized acellular surface was able to capture cells and also suppress platelet adhesion and fibrin clot deposition in a peptide density-dependent manner.

DOI： 10.1021/acsbiomaterials.0c00078

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Language：English   Publishing type：Research paper (scientific journal)  

Magnetic resonance angiography is an attractive method for the visualization of the cerebrovasculature, but small-sized vessels are hard to visualize with the current clinically approved agents. In this study, a polymeric contrast agent for the superfine imaging of the cerebrovasculature is presented. Eight-arm polyethylene glycol with a molecular weight of ≈17 000 Da conjugated with a Gd chelate and fluorescein (F-8-arm PEG-Gd) is used. The relaxivity rate is 9.3 × 10-3 m-1 s-1 , which is threefold higher than that of free Gd chelate. Light scattering analysis reveals that F-8-arm PEG-Gd is formed by self-assembly. When the F-8-arm PEG-Gd is intravenously injected, cerebrovasculature as small as 100 µm in diameter is clearly visualized. However, signals are not enhanced when Gd chelate and Gd chelate-conjugated 8-arm PEG are injected. Furthermore, small vasculature around infarct region in rat stroke model can be visualized. These results suggest that F-8-arm PEG-Gd enhances the MR imaging of cerebrovasculature.

DOI： 10.1002/mabi.201700391

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Language：Japanese   Publishing type：Research paper (scientific journal)   Publisher：Society of Polymer Science  

The amide functionality in the 2,5-diketopiperazine ring forms intermolecular hydrogen bonds between adjacent molecules that enable 2,5-diketopiperazines to form regulated higher-order supermolecular structures that are important in surface and coating engineering. In this study, we synthesized a cyclic(D-Tyr-D-Phe) peptide for the purpose of evaluating its self-assembly on glass or SUS316L, and the peptide's potential use in the design of antibacterial surfaces. A self-assembling 2,5-diketopiperazine nanostructure was observed and evaluated by atomic force microcopy (AFM). Cyclic(D-Tyr-D-Phe) formed a rod-like nanostructure on a glass surface. Compared to the synthetic H-D-Tyr-D-Phe-OH absorbed surface and the original material surfaces, self-assembling cyclic(D-Tyr-D-Phe) has the potential for antibacterial activity when adsorbed on surfaces. Finally, cyclic(D-Tyr-D-Phe) peptide did not show cell toxicity activity. The self-assembled cyclic(D-Tyr-D-Phe) has potential for use in the design of biomedical device surfaces with antibacterial activity.

DOI： 10.1295/koron.2017-0058

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Rapid in-situ re-endothelialization of coronary stents is one of the most effective approaches to inhibit late thrombosis and restenosis. Strut surfaces allowing excellent adhesion and migration of endothelial cells and endothelial progenitor cells may accelerate in-situ re-endothelialization. Here, a well-known endothelial cell adhesive peptide, Arg-Glu-Asp-Val (REDV), was directly immobilized onto metallic surfaces by means of single-step tyrosine oxidation with copper chloride (II) and hydrogen peroxide, which we recently reported as a new biomaterial modification technique. REDV immobilization on a 316L stainless steel plate improved endothelial cell adhesion and effectively suppressed platelet adhesion in vitro. In addition, a Co-Cr stent immobilized with Ac-Tyr-Gly-Gly-Gly-Arg-Glu-Asp-Val (Y-REDV) was implanted into a rabbit abdominal aorta. On 7 days postimplantation, 80% of the strut surface of the Y-REDV-immobilized stent was covered by a thin neointimal layer and was similar in appearance to native endothelium. Restenosis and late thrombosis were not observed in the Y-REDV-immobilized stent for 42 days. These findings suggest that direct immobilization of Y-REDV peptide onto metallic biomaterials by tyrosine oxidation is effective for promoting in-situ re-endothelialization in vascular stents. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 491-499, 2018.

DOI： 10.1002/jbm.a.36258

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：ELSEVIER SCIENCE BV  

Introduction: Salivary gland hypofunction, also known as xerostomia, occurs as a result of radiotherapy for head and neck cancer, autoimmune diseases, or aging. Xerostomia leads to oral health problems and thus affects the quality of life. Biological salivary gland tissue generated in vitro would provide an alternative mode of treatment for this disease.
Methods: To develop a novel method for modulating salivary gland tissue growth in vitro, we prepared a KP24 peptide-immobilized hydrogel sheet, wherein the peptide comprised repeating proline and lysine sequences, and evaluated the effect of this peptide on salivary gland tissue growth.
Results: We found that the KP24 peptide has the potential to enhance glandular tissue growth in vitro. This enhancement is associated with neurite outgrowth and increasing neural innervation.
Conclusion: KP24 peptide modified material would be a promising material for the modulation of salivary gland tissue growth in vitro. (C) 2016, The Japanese Society for Regenerative Medicine. Production and hosting by Elsevier B.V.

DOI： 10.1016/j.reth.2016.02.006

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DOI： 10.1295/koron2015-0030

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：ELSEVIER SCI LTD  

Researchers have attempted to develop efficient antithrombogenic surfaces, and yet small-caliber artificial vascular grafts are still unavailable. Here, we demonstrate the excellent patency of tissue-engineered small-caliber long-bypass grafts measuring 20-30 cm in length and having a 2-mm inner diameter. The inner surface of an acellular ostrich carotid artery was modified with a novel hetero-bifunctional peptide composed of a collagen-binding region and the integrin alpha 4 beta 1 ligand, REDV. Six grafts were transplanted in the femoral femoral artery crossover bypass method. Animals were observed for 20 days and received no anticoagulant medication. No thrombogenesis was observed on the luminal surface and five cases were patent. In contrast, all unmodified grafts became occluded, and severe thrombosis was observed. The vascular grafts reported here are the first successful demonstrations of short-term patency at clinically applicable sizes. (C) 2015 Elsevier Ltd. All rights reserved.

DOI： 10.1016/j.biomaterials.2015.04.031

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In vitro fabricated biological tissue would be a valuable tool to screen newly synthesized drugs or understand the tissue development process. Several studies have attempted to fabricate biological tissue in vitro. However, controlling the growth and morphology of the fabricated tissue remains a challenge. Therefore, new techniques are required to modulate tissue growth. RGD (arginine-glycine-aspartic acid), which is an integrin-binding domain of fibronectin, has been found to enhance cell adhesion and survival; it has been used to modify substrates for in vitro cell culture studies or used as tissue engineering scaffolds. In addition, this study shows novel functions of the RGD peptide, which enhances tissue growth and modulates tissue morphology in vitro. When an isolated submandibular gland (SMG) was cultured on an RGD-modified alginate hydrogel sheet, SMG growth including bud expansion and cleft formation was dramatically enhanced. Furthermore, we prepared small RGD-modified alginate beads and placed them on the growing SMG tissue. These RGD-modified beads successfully induced cleft formation at the bead position, guiding the desired SMG morphology. Thus, this RGD-modified material might be a promising tool to modulate tissue growth and morphology in vitro for biological tissue fabrication.

DOI： 10.1038/srep11468

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Language：Japanese   Publishing type：Research paper (scientific journal)   Publisher：SOC POLYMER SCIENCE JAPAN  

Polysaccharides such as alginate or chitosan have gained increasing attention for applications as soft biomaterials. In particular, alginate-Ca2+ hydrogel has been applied to food chemistry and as a tissue engineering scaffold. In this study, we prepared an alginate nanofiber without Ca2+ ions that utilizes self-assembling beta-sheet peptides as the cross-linker for the hydrogel. We designed this cross -linker from a sequence of hydrophilic and hydrophobic amino acids, such as (KLVFF)(4), and immobilized the self-assembled beta-sheet peptides to the -COOH group of the alginate to form cross links. We then investigated the molecular conformation of the self-assembled beta-sheet peptides in solution by circular dichroism and infrared spectroscopy.

DOI： 10.1295/koron.2015-0030

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A health monitoring system (HMS) involving a blood extraction device with a new type of hybrid biosensor comprising an enzyme and a semiconductor has recently been developed. A MOSFET was used as the transducer. The gate electrode was extracted from the MOSFET using a cable. Gold (Au)-plate-immobilized glucose oxidase (Go) was used as a biosensor and attached to the gate electrode. Go was immobilized on a self-assembled spacer combined with an Au electrode by the cross-link method using BSA as an additional bonding material. The electrode could be used to detect electrons generated by the oxidization of hydrogen peroxide produced by the reaction between Go and glucose using the constant electric current measurement system of the MOSFET-type hybrid biosensor. The sensitivities for the diluted whole blood and blood plasma were 61.4 and 171.2 V/(mol/L), respectively. The hybrid biosensor was useful for HMS. © 2013 VBRI press.

DOI： 10.5185/amlett.2012.6367

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Language：Japanese   Publishing type：Research paper (scientific journal)  

We designed water soluble copolymer for corrosion inhibition effects in aqueous enviroment. This copolymer was pursued for the goal of environmental adaptability. Water soluble copolymers were prepared from methoxypolyethylene glycol methacrylate (MPEG-MA) and acrylic acid units. We polymerized 5 to 65wt% of methoxypolyethylene glycol methacrylate and 95 to 35wt% of acrylic acid monomers by azobisisobutyronitrile (AIBN) and sodium peroxodisulfate in organic and water media, respectively. The solution containing these copolymers was evaluated inhibition of corrosion effect for steel materials. These results suggested that combination of MPEG-MA and acrylic acid in polymerizing in water media was most effective to inhibit corrosion for steel materials. It is important to contain 4 to 6 mol unit of ethylene oxide side chain in MPEG-MA. © 2012 The Society of Materials Science, Japan.

DOI： 10.2472/jsms.61.359

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Language：Japanese   Publishing type：Research paper (scientific journal)   Publisher：SOC POLYMER SCIENCE JAPAN  

RGDS peptide is a widely known cell-attachment peptide. In the present publication, the molecular design of molecules that incorporate two or more RGDS peptides and their conjugates with water-soluble polymers is reported. The cell attachment activity of these peptides was examined out by cell inhibition experiments. RGDS and (RGDS)(n) peptides in aqueous solutions were analyzed by molecular dynamics calculations. We discuss the relationship between the theoretical structural analysis and experimental conformation analysis obtained by CD spectra. The theoretical structural analysis and the experimental structural analysis were well in agreement. Furthermore, we discuss the activity of cell attachment and structure of peptide, from the viewpoint of cell attachment biomaterials.

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Language：Japanese   Publisher：The Japanese Society of Toxicology  

DOI： 10.14869/toxpt.39.1.0.P-7.0

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：INDERSCIENCE ENTERPRISES LTD  

The objective of this study was to prepare the complexes of 5-aminolevulinic acid (5-ALA) and iron oxide nanoparticles with different surface potentials and evaluate the imaging efficiency for mouse melanoma cells (B16 cells). The ferric and ferrous ions were co-precipitated in the presence of pullulan or the cationised and anionised derivatives to prepare iron oxide-pullulan nanoparticles (IOPNs). The surface potential of IOPN was changed by altering the mixing percentage of pullulan derivatives. The 5-ALA was mixed with the IOPN of different surface potentials to obtain IOPN-5-ALA complexes. When incubated with IOPN-5-ALA complexes, B16 cells exhibited the fluorescence of protoporphyrin IX (PpIX) bio-synthesised from 5-ALA internalised. The time profile of PpIX fluorescence percentage was greatly influenced by the surface potential of IOPN. This finding indicates that IOPN is a promising carrier for 5-ALA-based imaging of tumour cells.

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Language：Japanese   Publishing type：Research paper (scientific journal)   Publisher：SOC POLYMER SCIENCE JAPAN  

New-type branched beta-sheet peptides were designed using hydrophilic and hydrophobic amino acids. In the research, (Arg-Ala-Arg-Ala-Asp-Ala-Asp-Ala)(2) (RAD16) and RAD8 peptides were selected as typical sequences having high propensities for forming beta-strands. Thus, Lys residue was selected as a branching point, which introduced the spacer such as (D)Pro-Pro and Gly-Gly-Gly sequences. We investigated the stability of these peptides at several pH values and temperatures by measuring CD spectra and we studied the effects of self-assembling by observation of AFM. RAD16 and branched RAD8/8/16G took beta-sheet structures in water. However, RAD16, branched RAD8/8/16G and branched RAD16/16/8G built up nano fiber formations. Other random coil peptides in water were aggregated according to AFM observations. RAD16 and RAD8 peptides interact with each other and become more structured upon fiber formation.

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Language：Japanese   Publisher：The Society of Materials Science, Japan  

Biocompatible piezoelectric materials are becoming increasingly important for actuators and sensors in medical devices. In this paper, we highlighted on some perovskite oxides MgSiO₃, CaSiO₃, CaTiO₃ and CaZrO₃ which are biocompatible piezoelectric materials discovered by first-principles calculation in our previous studies. In order to verify their biocompatibility, the cytotoxicity of their isomeric oxides with the same components was examined as comparing with typical perovskite oxides Pb(Zr, Ti)O₃ and BaTiO₃. The fibroblast (L929) cells were cultured during 7 days and the effect of materials was evaluated by the half maximal inhibitory concentration (<i>IC</i>50). As a result, the colony formation assay indicated that BaTiO₃, CaTiO₃, MgSiO₃ and CaZrO₃ has the higher <i>IC</i>50 values. On the other hand, CaSiO₃ and Pb(Zr,Ti)O₃ shows the strong toxicity. The obtained <i>IC</i>50 values had a good correlation with the proliferation ratio in our previous studies.

DOI： 10.2472/jsms.58.943

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Other Link： https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-19760080/

Language：Japanese   Publishing type：Research paper (scientific journal)  

Biocompatible piezoelectric materials are becoming increasingly important for actuators and sensors in medical devices. In this paper, we highlighted on some perovskite oxides MgSiO3, CaSiO3, CaTiO3 and CaZrO3 which are biocompatible piezoelectric materials discovered by first-principles calculation in our previous studies. In order to verify their biocompatibility, the cytotoxicity of their isomeric oxides with the same components was examined as comparing with typical perovskite oxides Pb (Zr, Ti)O3 and BaTiO3. The fibroblast (L929) cells were cultured during 7 days and the effect of materials was evaluated by the half maximal inhibitory concentration (IC50). As a result, the colony formation assay indicated that BaTiO3, CaTiO3, MgSiO3 and CaZrO3 has the higher IC50 values. On the other hand, CaSiO3 and Pb (Zr, Ti)O3 shows the strong toxicity. The obtained IC50 values had a good correlation with the proliferation ratio in our previous studies. ©2009 The Society of Materials Science, Japan.

DOI： 10.2472/jsms.58.943

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：VSP BV  

The adhesion and proliferation of human adipo-stromal cells was investigated for poly(ethylene terephthalete) (PET) films of two-dimensional (2D) substrate and non-woven fabrics of three-dimensional (3D) substrate after seeding at the same cell density and culturing at the same medium volume to surface area of the substrates ratio. When seeded by a static seeding method, more cells adhered on the film than on the non-woven fabric. However, the number ratio of cells proliferated to those initially adhered was similar. For the non-woven fabric, cell proliferation was enhanced by an agitated culture method. NaOH treatment introduced carboxyl groups into the surface of substrates, irrespective of the substrate type. Cell adhesion of a peptide (Arg-Gly-Asp, RGD) was chemically immobilized through the carboxyl groups on the non-woven fabric surface at a density of 10 pmol/cm(2). RGD immobilization significantly increased the number of cells adhered after the agitated seeding method, but did not affect the cell proliferation. Phosphorylation of focal adhesion kinase (FAK) was also enhanced by the RGD immobilization on the PET non-woven fabrics. (C) Koninklijke Brill NV, Leiden, 2009

DOI： 10.1163/156856209X426600

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：VSP BV  

The objective of this study is to investigate the adhesion of human adipo-stromal cells on self-assembled monolayers (SAM) with different surface densities and gradients of Arg-Gly-Asp (RGD) peptide. The different densities and gradients of carboxyl groups on the SAM surface were prepared by a SAM exchange technique, and then RGD was chemically immobilized to allow the RGD surface density on the SAM to change over the range of 2.3-8.9 ng/cm(2) (3.0-12 pmol/cm(2)). The spreading area and survival percent of cells increased as the density of RGD immobilized on the SAM became high, whereas no dependence of the RGD density on the number of cells adhered was observed. The survival percent of cells tended to increase with an increase in the RGD immobilization for the carboxyl group-gradient SAM. This finding suggests the possibility that the cell adhesion can be regulated by controlling the RGD density or gradient of cell substrate. (C) Koninklijke Brill NV, Leiden, 2009

DOI： 10.1163/156856209X416502

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：ELSEVIER SCI LTD  

The complement system is strongly activated by surfaces carrying nucleophilic groups, such as hydroxyl (OH) groups, and triggered by deposition of complement protein fragment, Ob. Surfaces carrying amino groups, the other representative nucleophilic group, are expected to be potential activators of the complement system through the alternative pathway. Few studies thus far have examined the potential of artificial materials carrying amino groups in activating the complement system. In this study, we employed a self-assembled monolayer (SAM) of 11-amino-1-undecanethiol (NH2-SAM) and a polyethyleneimine (PEI)-coated surface as model surfaces to study interactions between amino groups and serum complement pathway. SAMs of 11-mercaptoundecanol (OH-SAM) and 1-dodecanethiol (CH3-SAM) were used as control surfaces, respectively. Although much protein was adsorbed from serum solutions on the two types of amino surfaces, amounts of Ob deposition were much less than those observed on OH-SAM. Amounts of C3a released on the amino surfaces were same levels as that of CH3-SAM, but significantly smaller than that on OH-SAM. These facts suggest that the nucleophilic amino groups on NH2-SAM and PEI-coated surfaces do not directly activate the alternative pathway, but the protein adsorbed layers formed on amino surfaces activate it, but to an extent much smaller than that on OH-SAM. In addition, we found no deposition of C1q molecules on the amino surfaces, suggesting that these surfaces fail to activate the classical pathway. However, more careful studies are needed to conclude it, because it is known that C1q is only transiently detected at typical classical activation interfaces. (c) 2007 Elsevier Ltd. All rights reserved.

DOI： 10.1016/j.biomaterials.2007.10.005

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Language：Japanese   Publishing type：Research paper (scientific journal)   Publisher：SOC POLYMER SCIENCE JAPAN  

Polysaccharides such as alginate or chitosan have gained increasing attention for applications as soft biomaterials. In particular, alginate-Ca(2+) hydrogels have been applied to food chemistry and as tissue engineering scaffold. In contrast, alginate-metal ion hydrogels are toxic to liver cells and tissue. In this study, we prepared an alginate hydrogel without Ca(2+) ions that utilizes self-assembling beta-sheet peptides as the cross-linker for the hydrogel. We designed this cross-linker from a sequence of hydrophilic and hydrophobic amino acids, such as (FEFK)(n) where n = 2-5, and the self-assembled beta-sheet peptides immobilized to the -COOH group of the alginate to form cross-links. We then investigated the molecular conformation of the self-assembled beta-sheet peptides in solution and in solid-state by circular dichroism and Fourier-trans form infrared spectroscopy and determined the optimal gelating conditions of the alginate. Here, we present a novel method of using self-assembling beta-sheet peptides as a cross-linker for a new alginate hydrogel that has potential use in drug delivery materials and as tissue engineering scaffolds.

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Language：English   Publishing type：Research paper (international conference proceedings)   Publisher：SPIE-INT SOC OPTICAL ENGINEERING  

Hemodialysis for chronic renal failure is the most popular treatment method with artificial organs. However, hemodialysis patients must continue the treatment throughout their life, the results of long term extracorporeal dialysis, those patients develop the various complications and diseases, for example, dialysis amyloidosis etc. Dialysis amyloidosis is one of the refractory complications, and endotoxin is thought to be the most likely cause of it, recently. Endotoxin is one of the major cell wall components of gram-negative bacteria, and it has various biological activities. In addition, it is known that a mount of endotoxin exists in living environment, and medicine is often contaminated with endotoxin. When contaminated dialyzing fluids are used to hemodialysis, above-mentioned dialysis amyloidosis is developed. Therefore, it is important that the detection and removal of endotoxin from dialyzing fluids. Until now, the measurement methods using Limulus Amebosyte Lysate (LAL) reagent were carried out as the tests for the presence of endtoxin. However, these methods include several different varieties of measurement techniques. The following are examples of them, gelatinization method, turbidimetric assay method, colorimetric assay method and fluoroscopic method. However, these techniques needed 30-60 minutes for the measurement. From these facts, they are not able to use as a "real-time endotoxin detector". The detection of endotoxin has needed to carry out immediately, for that reason, a new detection method is desired. In this research, we focused attention to adsorption reaction between ε-polylysine and endotoxin, ε-polylysine has the structure of straight chain molecule composed by 25-30 residues made by lysine, and it is used as an antimicrobial agent, moreover, cellulose beads with immobilized ε-polylysine is used as the barrier filter for endotoxin removal. The endotoxin is adsorbed to immobilized ε-polylysine, as the result of this reaction, the mass incrementation is occurred, and the existence of endtoxin can be detected immediately, by using of Quartz Crystal Microbalance (QCM). In this report, the immobilization of ε-polylysine onto the Au and Si substrate and its adsorptive activity are described. We use X-ray Photoelectron Spectroscopy (XPS) to confirm the ε-polylysine immobilization, and the adsorptive activity of immobilized ε-polylysine is measured by AFM and QCM. This molecular adsorption type endotoxin sensor aims to the realization of "real-time endotoxin detection system".

DOI： 10.1117/12.759459

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：VSP BV  

DOI： 10.1163/156856207781367756

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：ELSEVIER SCIENCE BV  

The objective of this study is to prepare a novel gene carrier from pullulan, a polysaccharide with an inherent affinity for the liver, and evaluate the feasibility in gene transfection. Various amounts of spermine were chemically introduced into pullulan with molecular weights of 22,800, 47,300, and 112,000 to prepare cationized pullulan derivatives with different percentages of spermine introduced. Each cationized pullulan derivative was complexed with a plasmid DNA at various ratios and applied to HepG2 cells for in vitro gene transfection. The level of gene expression depended on the percent spermine introduced of cationized pullulan derivatives and the molecular weight of pullulan. However, when compared at the complexation molar ratio of pullulan derivative to the plasmid DNA, the expression level became maximum around the ratio of 10(2), irrespective of the pullulan molecular weight. Pre-treatment of cells with asialofetuin of asialoglycoprotein receptor ligand decreased the level of gene expression by the complexes. The cationized pullulan derivative with an appropriate physicochemical character is a promising non-viral carrier which promotes the receptor-mediated internalization of plasmid DNA and consequently enhances the expression level. (c) 2007 Elsevier B.V. All rights reserved.

DOI： 10.1016/j.jconrel.2007.01.005

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：SPRINGER  

Poly(proline) and random copolypeptide composed of Pro and Ala residues were synthesized, and their solution properties and molecular conformation were investigated. Aqueous solutions of the polypeptide were irradiated with gamma-rays above the transition temperature. It was shown that the transition temperature of the aqueous solution of the copolypeptide is influenced by Ala-residue content and gamma-ray irradiation.

DOI： 10.1007/s00289-006-0666-4

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Language：English   Publishing type：Research paper (international conference proceedings)   Publisher：SPIE-INT SOC OPTICAL ENGINEERING  

Patients with renal failure become not able to expel the waste product, and they accumulate the toxic products for themselves. They therefore must use the hemodialysis to weed out the metabolic decomposition product. Hemodialysis for chronic renal failure is the most popular treatment method with artificial organs. However, hemodialysis patients must continue the treatment throughout their life, the results of long term extracorporeal dialysis, those patients develop the various complications and diseases, for example, dialysis arnyloidosis etc. Dialysis amyloidosis is one of the refractory complications, and the prevention of this complication is important. Recently, endotoxin is thought to be the most likely cause of the complication.Endotoxin is one of the :major cell wall components of gram-negative bacteria, and it forms the complex with proteins and lipopolysaccharide (LPS). It has various biological activities, e.g. attack of fever, when it gets mixed into human blood. In addition, it is known that large amount of endotoxin exists in living environment, and medicine is often contaminated with endotoxin. When contaminated dialyzing fluids are used to hemodialysis, above-mentioned dialysis amyloidosis is developed Therefore, it is important that the detection and removal of endotoxin from dialyzing fluids. Until now, the measurement methods using Limulus Amebosyte Lysate (LAL) reagent were carried out as the tests for the presence of endotoxia. However, these methods include several different varieties of measurement techniques. The following are examples of them, gelatinization method, turbidimetric assay method, colorimetric assay method and fluoroscopic method. However, these techniques needed 30-60 minutes for the measurement. From these facts, they are not able to use as a "real-time endotoxin detector". The detection of enclotoxin has needed to carry out immediately, for that reason, a new "real-time" detection method is desired.We focused attention to adsorption reaction between, epsilon-polylysine and enclotoxin. epsilon-polylysine has the structure of straight chain molecule composed by 25-30 residues made by lysine, and it is used as an antimicrobial agent, moreover, cellulose beads with immobilized E-polylysine is used as the barrier filter for enclotoxin removal. Therefore, it is expected that the enclotoxin be adsorbed to the immobilized epsilon-polylysine onto the probe. As the result of this reaction, the mass of the probe is increased, and enclotoxin can be detected by using of Quartz Crystal Microbalance (QCM). In our previous research, we. have already acquired the proteins immobilization technique onto Au and Si surface. In this report, the proposal of molecular adsorption type endotoxin detection system, and the immobilization of E-polylysine onto the probe are described. We use X-ray Photoelectron Spectroscopy (XPS) to confirm the F-polylysine immobilization, and the adsorptive activity of immobilized E-polylysine is measured by XPS and AFM. The purpose of this study is to bring about the realization of "Real-time enclotoxin detection system".

DOI： 10.1117/12.715985

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Language：English   Publishing type：Research paper (international conference proceedings)   Publisher：IEEE  

When renal failure patients undergo the hemodialysis, the endtoxin (ET) that is the bacterial toxin that causes the symptom such as generation of heat is measured as one of the pollution markers. Presently, because the high performance filtration equipment develops, the amount of ET in dyalysate water and solution was achieved under the ET standard value set by Japanese Society for Dialysis Therapy etc. However, the ET measurement is obligated to the moon a couple of degrees at the dialysis center etc. the measurement is needed for 30 minutes or more since it use batch method and rely on the chemical reaction.In this study, we paid attention to the polyamino said named epsilon-Polylisine (epsilon-PL) that can select and adsorb ET and the mass measurement using Quartz crystal microbalance method (QCM). We aim to the development of the real-time ET measuring system using these. If it is achieved, the amount of ET can be monitored while blood is dialyzed. Moreover, it seems that treatment that discontinues dialyzing can be taken when it exceeds the reference value.

DOI： 10.1109/MHS.2007.4420860

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Language：English   Publishing type：Research paper (international conference proceedings)   Publisher：TRANS TECH PUBLICATIONS LTD  

Therapeutic angiogenesis by localized delivery of angiogenic factors is a promising approach to treat patients with cardiovascular disease and to engineer large tissues. Vascular endothelial growth factor (VEGF) is the most common and biologically active form of the VEGF family. which acts as a mitogen to endothelial cells and is capable of spec fie binding to heparin. However, when VEGF is administered via bolus injection, it can be widely distributed, and its concentration is likely to be within the effective window for only a short time period due to rapid degradation. Delivery of angiogenic factors, using controlled drug delivery strategies, offers great potential to promote angiogenesis at a specific site while reducing the unwanted side effects that may occur with systemic delivery. We now report on the sustained release of VEGF from alginate gels, modified with a heparin-binding peptide. Briefly, a small peptide with the sequence of G(5)K(PA)FAKLAARLYRKA, which is known to specifically interact with heparin, was chemically conjugated to alginate, and the peptide-modified alginate formed gels after mixing with heparin and VEGF. The release rate of VEGF from the gels slowed in vitro for over 45 days, compared with release from non-modified alginate gels. This result suggests potential applications of alginate gels in promoting angiogenesis for therapeutic purposes, as well as for tissue engineering.

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Publisher：The Society of Life Support Engineering  

DOI： 10.5136/lifesupport.18.Supplement_87

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Publishing type：Research paper (international conference proceedings)  

Cell growth efficiencies in various bioreactors are not sufficient for in vitro tissue regeneration. Recently rapid vascular tissue regeneration using plasmid DNA coding vascular endothelial growth factors (VEGF) have been reported. We newly prepared PLLA porous scaffold containing pVEGF/polycations polyplexes for direct gene transfection to HUVEC cells on the scaffold resulting in the enhanced cell growth and tissue regeneration. The VEGF expression in HUVECs was monitored using real time PCR and its correlation with the cell growth rate was studied.

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Language：Japanese   Publishing type：Research paper (international conference proceedings)  

Adhesion of human adipo-stromal cells was investigated for substrates of which surface was modified by immobilization of a cell adhesion peptide RGDS. The substrate surface was gold-coated and modified by a self-assemble technique with different alkanethiols, while the RGDS was chemically immobilized. The modified surface was characterized by water wettability and Electron Spectroscopy for Chemical Analysis measurements. The amount of RGDS immobilized was determined by using radioiodinated RGDS. Cell adhesion was evaluated in terms of the area and number of cells adhered. The highest cell spreading was observed for the RGDS-immobilized substrate with 11-mercaptoundecanoic acid.

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Language：English   Publishing type：Research paper (international conference proceedings)  

As the population ages, health management will be one of the important issues. The development of a safe medical machine based on MEMS technologies for the human body will be the primary research project in the future. We have developed the glucose sensor, as one of the medical based devices, for use in the Health Monitoring System (HMS). HMS is the device that continuously monitors human health conditions. For example, blood is the monitoring target of HMS. The glucose sensor specifically detects the glucose levels of the blood and monitors the glucose concentration as the blood sugar level. This glucose sensor has a "separated Au electrode," which immobilizes GOx. In our previous work, GOx was immobilized onto Au electrode by the SAMs (Self-Assembled Monolayer) method, and the sensor, using this working electrode, detected the glucose concentration of an aqueous glucose solution. In this report, we used a Pt electrode, which immobilized GOx, as a working electrode. Au electrode, which was used previously, was dissolved by the application of current in the presence of chloride ions. Based on the above-mentioned fact, a new working electrode, which immobilized GOx, was produced using Pt, which did not possess such characteristics. These Pt working electrodes were produced using the covalent binding method and the cross-link method, and both the electrodes displayed a good sensing property. In addition, the electrode using glutaraldehyde (GA) and bovine serum albumin (BSA) as crosslinking agents was produced, and it displayed better characteristics as compared with those displayed by the electrode that used only GA. Based on the above-mentioned techniques, the improvement in performance of the sensor was confirmed.

DOI： 10.1117/12.582346

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：SPRINGER  

Several types of poly(Gly-co-Pro) and poly(Ala-co-Pro) were synthesized. Their molecular conformations and thermosensitive properties were investigated in an attempt to find new thermosensitive materials. These polypeptides were assumed the polyproline-II structure in the temperature range of 20 to 80 degrees C. They also exhibited cloud points in light transmittance, indicating the phase transition. The transition temperatures increased with decrease in hydrophobicity of the polypeptides, that is, in the order poly(proline), poly(Ala-co-Pro) and poly(Gly-co-Pro).

DOI： 10.1007/s00289-005-0384-3

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：SPRINGER  

Proline oligopeptides composed of 13, 6 and 4 Pro residues were synthesized by solid-phase procedure. The circular dichroism spectra showed that the proline oligopeptides form the polyproline-II helix in water and in trifluoroethnol. However, it was shown that the oligopeptide composed of 13 Pro residues remarkably presents conformational transition from the polyproline-II to the polyproline-I helix in methanol and 1-propanol. This result is the first spectrum evidence that proline oligopeptides can form the polyproline-I helix in pure aliphatic alcohol such as methanol and 1-propanol. It was also shown that the propensity forming polyproline-I helix is more favorable in 1-propanol than in methanol, and also that the longer the chain-length is, the greater the stability of the polyproline-I helix is. Such a chain-length dependency of the conformational stability is also supported by the theoretical calculation using molecular mechanics.

DOI： 10.1007/s00289-004-0317-6

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：VSP BV  

The objective of this study is to investigate the proliferation and differentiation of stromal cells derived from human adipose tissues cultured on substrates with different surface properties. In addition, a similar investigation was performed on cells proliferated in different concentrations of basic fibroblast growth factor (FGF-2). The culture substrates include several polymer films with different water wettabilities, glass or a cell-culture plate, and that coated with collagen type I or IV, gelatin and FGF-2. The proliferation profiles of cells were influenced by the type of culture substrate and the growth factor concentration. A larger number of proliferated cells was observed for substrates with a water contact angle around 80 degrees, while the cell number was significantly larger for every protein-coated substrate. The rate of cell proliferation became maximal at a FGF-2 concentration of 1000 ng/ml. The FGF-2 concentration used for cell proliferation affected the differentiation profile of cells proliferated. Stromal cells, proliferated in 1 ng/ml FGF-2, were osteogenically differentiated to the strongest and fastest extent among those in other growth factor doses. The alkaline phosphatase (ALP) activity of cells increased with the increased cell number, although the activity per cell was identical, irrespective of the substrate type. The strongest adipogenic differentiation was observed for cells proliferated in 1000 ng/ml FGF-2 and the differentiation induction was maintained for a long time period. No clear dependence of the cell number on adipogenesis was observed. These findings indicate that the proliferation and differentiation of human adipose tissue-derived stromal cells are influenced by the culture substrate and the concentration of FGF-2 used for proliferation.

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：ELSEVIER SCIENCE BV  

Storage and release of endogenous growth factors by the extracellular matrix (ECM) are important biological events that control tissue homeostasis and regeneration. The interaction between basic fibroblast growth factor (bFGF) and heparan sulfate proteoglycans has been extensively studied and used as a prototype model of such a system, while the lower affinity of fibrillar type I collagen for bFGF has generally been considered biologically insignificant. However, our present investigation revealed that bFGF spontaneously interacts with type I collagen solution and sponges under in vitro and in vivo physiological conditions, and is protected from the proteolytic environment by the collagen. bFGF incorporated in a collagen sponge sheet was sustainedly released in the mouse subcutis according to the biodegradation of the sponge matrix, and exhibited local angiogenic activity in a dose-dependent manner. Intramuscular injection of collagen microsponges incorporating bFGF induced a significant increase in the blood flow in the murine ischemic hindlimb, which could never have been attained by bolus injection of bFGF. These results suggest the significance and therapeutic utility of type I collagen as a reservoir of bFGF. (C) 2004 Elsevier B.V. All rights reserved.

DOI： 10.1016/j.jconrel.2004.07.008

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：MARY ANN LIEBERT INC  

This study is an investigation of the proliferation and differentiation of bone marrow stromal cells (BMSCs) on film substrates with different surface properties. Films of noncharged polymers with several water wettabilities; cell culture plates coated with collagen type I or IV, gelatin, or basic fibroblast growth factor (bFGF); and glass were used. BMSCs isolated from rat bone marrow were cultured on the various substrates in medium with dexamethasone [Dex(+)] or without dexamethasone [Dex(-)] to assess cell proliferation and differentiation. The number of proliferated BMSCs depended on the water wettability of substrates, although the cell number was greater in Dex(-) medium than in Dex(+) medium. Protein-coated substrates exhibited a high proliferation rate compared with noncoated substrates. Alkaline phosphatase (ALP) activity increased with increasing cell number, whereas ALP activity per cell correlated well with cell number. When cultured in Dex(+) medium containing bFGF or on culture plates coated with bFGF, BMSC proliferation tended toward enhancement with an increase in the amount of bFGF added in solution form, whereas it did not depend on the amount of bFGF in coated form. On the other hand, ALP activity and calcium content of BMSCs became maximal with bFGF coated at about 1 x 10(3) to 2 x 10(3) ng, in contrast to bFGF in solution form. Irrespective of the amount of bFGF, ALP activity and calcium content levels for bFGF in coated form were higher than for bFGF in solution form. It is concluded that the type of culture substrate and the manner of addition of bFGF affect the proliferation and differentiation of BMSCs.

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Language：English   Publisher：WILEY-V C H VERLAG GMBH  

Tissue engineering aims to create new tissues and organs to replace those lost to disease, trauma, or congenital defects. However, considerable instruction must be given to the cells forming these new tissues if one is to create a tissue structurally and functionally similar to the native tissue. Materials are ideal for the local presentation of various peptides and proteins that can bind to cell surface receptors and regulate cell function. An overview of techniques to present peptides and proteins from materials, and the utility of these systems in engineering tissues in vitro and in vivo is presented.

DOI： 10.1002/adma.200300383

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Language：English   Publishing type：Research paper (international conference proceedings)  

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Other Link： http://orcid.org/0000-0002-6683-4513

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Language：English   Publishing type：Research paper (international conference proceedings)   Publisher：SPIE-INT SOC OPTICAL ENGINEERING  

As the population ages, the management of the health is one of the important problems. Development of the harmless medical machine based on MEMS technologies for human body will be the mainly research projects in the future. Before now, we developed the glucose sensor for using at Health Monitoring System (HMS) as one of the medical device based on micromechatronics. HMS is the device that monitors human health conditions continuously. For example, the monitoring target of HMS is the blood. The whole blood contains the manifold health index markers, and it is very important to measure them in the health care. Glucose sensor specifically detects the glucose of the blood, and it monitors the glucose concentration as blood sugar level. This glucose sensor had 'separated Au electrode' which immobilized GOx. By utilizing this style, it becomes possible that the sensor part is easily miniaturized. In our previous work, GOx was immobilized onto Au electrode by using of SAMs (Self-Assembled Monolayor) method, and the sensor using this working electrode detected the glucose concentration of glucose aqueous solution. Furthermore, the miniaturization of Au electrode was realized.
In this report, glucose sensor which immobilized GOx using the cross-link method was produced, and the performance comparison with the sensor using SAMs method was carried out. And we carried out operation confirmation of produced glucose sensor using dilution human serum and whole blood. In addition, the cholesterol sensor which immobilized cholesterol oxidase (ChOx), which specifically detects the cholesterol on the blood, onto separated Au electrode by cross-link method was produced. The immobilization of the ChOx was evaluated from the spectra of XPS, and the performance as a sensor was evaluated.

DOI： 10.1117/12.522815

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：INT AMER ASSOC DENTAL RESEARCHI A D R/A A D R  

It is widely assumed that coupling the degradation rate of polymers used as cell transplantation carriers to the growth rate of the developing tissue will improve its quantity or quality. To test this hypothesis, we developed alginate hydrogels with a range of degradation rates by gamma-irradiating high-molecular-weight alginate to yield polymers of various molecular weights and structures. Decreasing the size of the polymer chains increased the degradation rate in vivo, as measured by implant retrieval rates, masses, and elastic moduli. Rapidly and slowly degrading alginates, covalently modified with RGD-containing peptides to control cell behavior, were then used to investigate the effect of biodegradation rate on bone tissue development in vivo. The more rapidly degrading gels led to dramatic increases in the extent and quality of bone formation. These results indicate that biomaterial degradability is a critical design criterion for achieving optimal tissue regeneration with cell transplantation.

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：ELSEVIER SCI LTD  

Phosphorylcholine (PC)-endcapped oligomer and block co-oligomer were prepared by employing a photoiniferter-based quasi-living polymerization technique. The designed oligomer had a PC polar head group attached to an alkylene chain at one end of the molecule and an oligo(styrene) (oligoST) segment at the other end. In the co-oligomer, an oligo(N,N-dimethylacrylamide) (oligoDMAAm) segment was inserted between both ends of the oligomer mentioned above. Surface coating of these amphiphilic substances, using an appropriate coating procedure, resulted in a very hydrophilic characteristic, suggesting that the oligoST anchored on the substrate and the PC polar head group was exposed to or located on the outer coating layer. Non-cell adhesivity in serum-containing medium was observed, while slightly reduced protein adsorption was observed. Thus, PC-endcapped oligomer and block co-oligomer appear to function as a biocompatible coating. (C) 2003 Published by Elsevier Science Ltd.

DOI： 10.1016/S0142-9612(03)00344-2

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Language：English  

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Language：Japanese   Publisher：高分子刊行会  

CiNii Books

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Language：English   Publishing type：Research paper (international conference proceedings)   Publisher：SPIE-INT SOC OPTICAL ENGINEERING  

A lot of researches and developments of medical device based on micromechatronics are performed. We develop the glucose sensor for using at HMS (Health Monitoring System) as one of the devices of those. In the development of a glucose sensor, it is necessary that the efficient hybridization between enzyme (glucose oxidase : GOD) as receptor and semiconductor as transducer. We devised the biosensor which used GOD immobilized gold plate as a gate electrode of the MOSFET. By utilizing this type, it becomes possible that the sensor part is easily miniaturized. When the glucose was detected using this sensor, there was a signal with concentration dependence. Therefore, it was confirmed that the prototype glucose sensor operated as a biosensor. From the above results, MOSFET type biosensor with gate electrode which immobilized GOD produced in this paper seems to be enough available as a glucose sensor. For miniaturization of the system, we examined the relationship between size of the working electrode and detection sensitivity. Consideration of the electrode area was not needed to detect the signal sensitively from the glucose sensor. And when we changed glass reference electrode to Au electrode, the enough sensitivity was obtained, too.

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Language：Japanese   Publisher：日本DDS学会  

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Language：Japanese   Publisher：(公社)日本薬学会  

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：TAYLOR & FRANCIS LTD  

This paper investigates the controlled release of hepatocyte growth factor (HGF) by biodegradable gelatin hydrogels and their HGF-induced angiogenic effect. Hydrogels of different degradabilities were prepared through chemical crosslinking gelatin with varied amounts of glutaraldehyde. When the gelatin hydrogels were radioiodinated and subcutaneously implanted into the back of mice, the remaining radioactivity of the hydrogels decreased with time. However, the remaining period became longer when the concentration of glutaraldehyde used for hydrogel preparation increased. Following implantation of gelatin hydrogels incorporating I-125-labeled HGF, the HGF radioactivity retained in the mouse subcutis for longer time periods as the glutaraldehyde concentration becomes higher. The time profile of HGF remaining in every, gelatin hydrogel was in good accordance with that of hydrogel degradation, indicating HGF release as a result of hydrogel biodegradation. The gelatin hydrogel incorporating HGF histologically induced angiogenic change around the implanted hydrogel. Gelatin hydrogels incorporating 5 and 10 mug HGF significantly enhanced the number of capillaries newly formed around the implanted site. This was in marked contrast to free HGF of same dose form and HGF-free, empty gelatin hydrogel. The gelatin hydrogel incorporating HGF induced VEGF around the implanted site. In vitro bioassay revealed that HGF molecules interacting with gelatin, still exhibited the biological activity. The interacted HGF would be released from gelatin hydrogels only when they were degraded to generate water-soluble gelatin fragments. It is possible that the HGF associating gelatin fragments of bioactivating, results in induced angiogenic effect.

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：MARY ANN LIEBERT INC PUBL  

De novo adipogenesis at the implanted site of a basement membrane extract (Matrigel) was induced through controlled release of basic fibroblast growth factor (bFGF). bFGF was incorporated into biodegradable gelatin microspheres for its controlled release. When the mixture of Matrigel and bFGF-incorporated gelatin microspheres was implanted subcutaneously into the back of mice, a clearly visible fat pad was formed at the implanted site 6 weeks later. Histologic examination revealed that the de novo formation of adipose tissue accompanied with angiogenesis was observed in the implanted Matrigel at bFGF doses of 0.01, 0.1, and 1 mu g/site, the lower and higher doses being less effective. The de novo formation induced by the bFGF-incorporated microspheres was significantly higher than that induced by free bFGF of the same dose. The mRNA of a lipogenesis marker protein, glycerophosphate dehydrogenase, was detected in the formed adipose tissues, biochemically indicating de novo adipogenesis. Free bFGF, the bFGF-incorporated gelatin microspheres, or Marigel alone and bFGF-free gelatin microspheres with or without Matrigel did not induce formation of adipose tissue. This de novo adipogenesis by mixture of Matrigel and the bFGF-incorporated gelatin microspheres will provide a new idea for tissue engineering of adipose tissue.

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：VSP BV  

The objective of this study is to investigate the influence of gelatin complexation on the biological activity of basic fibroblast growth factor (bFGF) and its resistance to trypsin digestion. When bFGF was mixed at 37 degrees C with acidic gelatin with an isoelectric point (IEP) of 5.0, the activity to promote in vitro proliferation of BI-IK cells became lower compared with that of free bFGF, in contrast to mixing with the basic gelatin with an IEP of 9.0. A maximum reduction in the bFGF activity was observed fur thr bFGF-gelatin complex prepared at a mixing molar ratio of 1/1. The bFGF activity of cell proliferation reduced at the initial period after mixing with the acidic gelatin at 37 degrees C, followed by no substantial change. Complexation with the acidic gelatin at 4 degrees C had no influence on the bFGF activity, irrespective of the bFGF/gelatin ratio and complexation time. The biological activity of bFGF was reduced by the trypsin treatment, but the reduced extent was suppressed through gelatin complexation at 37 degrees C. in an electrophoresis study, the protective effect of gelatin complexation on thr trypsin digestion was also confirmed in terms of the molecular weight loss. It is possible that the complexing gelatin covers bFGF molecules, resulting in suppression of their interaction with the cell surface receptor as well as protection from their enzymatic attack.

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：ELSEVIER SCI LTD  

This article reports a novel preparation method of poly(ethylene glycol) (PEG)-polystyrene (PST) amphiphilic block copolymers with well-defined block lengths by using photopolymerization of an iniferter, benzyl N,N-diethyldithiocarbamate. PEG macroiniferters, which were prepared by end-capping of PEG monomethyl ethers with benzyl N,N-diethyldithiocarbamate group at one end, were irradiated with UV light in the presence of styrene (ST). NMR analyses showed that the PST block was chain-extended from the PEG block, resulting in the preparation of PEG-PST block copolymers. The number-average molecular weights of the copolymers increased almost linearly with irradiation time, light intensity, and concentration of ST. The polydispersities of the copolymers remained relatively small throughout the reaction (M-w/M-n, approximate to 1.3). The composition of two PEG-PST block copolymers thus obtained was as follows: PEG (M-n; 1.9 x 10(3) g mol(-1))-PST (3.0 x 10(3) g mol(-1)) and PEG (4.9 x 10(3) g mol(-1))-PST (2.6 x 10(3) g mol(-1)). These copolymers were coated onto a poly(ethylene terephthalate) film surface. X-ray photoelectron spectroscopy analyses and water wettability measurements showed that the PST block was enriched at the outermost layer as cast in air, whereas upon immersion into water, the PEG block was oriented toward water. Enhanced wettability was observed for the diblock copolymer with a higher PEG content. Significantly reduced cell adhesion was observed on both the coated surfaces. Thus, the PEG-PST block copolymer may function as a cell adhesion-resistant coating which reduced cell-substrate interaction. (C) 1999 Published by Elsevier Science Ltd. All rights reserved.

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Language：English   Publishing type：Research paper (international conference proceedings)   Publisher：SPRINGER  

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：AMER CHEMICAL SOC  

This paper reports a surface process technology that simultaneously enables the formation of a hydrogel and its fixation on the surface of gold substrates. The technology utilizes two different chemical reactions of thiols: oxidation-induced disulfide bond formation and chemisorption on gold. Thiol-derivatized poly (vinyl alcohol)s (SH-PVAs), which were synthesized by radical copolymerization of vinylbenzyl N,N-diethyldithiocarbamate and vinyl acetate, followed by alkaline hydrolysis, were coated on gold surfaces and then air-dried at room temperature. X-ray photoelectron spectroscopy (XPS) analyses and water contact angle measurements of a gold surface before and after treatment with SH-PVA provided evidence that a hydrogel was formed and fixed on the gold surface. Atomic force microscopic (AFM) observations showed that the gel was tightly fixed even after thorough washing with hot water. Surfaces with regional hydrogelation, which were prepared on a gold-deposited micropatterned polymer, were visualized by cellular patterning under a phase-contrast microscope and by fluorescein-labeled protein immobilization under a fluorescent microscope.

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：Springer Verlag  

Theoretical conformational analysis was carried out for four disulfide-linked tetrapeptides Ac-Cys-Pro-D-Xaa-Cys-NHMe(D-Xaa= D-Val, D-Phe, D-Leu, and D- norleucine) using ECEPP and optimization procedure for investigating how the stabilities of the β-bend conformation at D-Xaa-Pro portion are affected by branching and bulkiness of hydrophobia side-chain groups of D-Xaa residue. Calculated results indicate that cyclic Ac-Cys-Pro-D-Xaa-Cys-NHMe commonly have a dominant character taking type II β-bend at the Pro-D-Xaa portion and also show fairly good agreement with experimental results of the NMR spectroscopy for a tetrapeptide Ac-Cys-Pro-D-Val-Cys- NH2. It is shown that the Ala-residue approximation is also a reasonable method to investigate the basic local conformational character of the Cys-Pro-D-Xaa-Cys sequence as well as that of the Cys-Pro-Xaa-Cys one. Moreover, it is suggested that the disulfide-linked tetrapeptides Ac-Cys-Pro-D-Xaa-Cys-NHMe are good candidates of the simple standard molecules for investigating the spectroscopic characters related to type II β- bend conformations.

DOI： 10.1007/s002890050410

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：NATURE PUBLISHING GROUP  

Theoretical conformational analysis was carried out on four cyclic tetrapeptides Ac-Cys-Pro-Xaa-Cys-NHMe (Xaa=Val, Phe, Leu, and norleucine) using Empirical Conformation Energy Program for Peptides (ECEPP) and optimization procedure for investigating the effects of differences in the hydrophonbic side-chain groups of Xaa residue on the beta-bend conformation at the Xaa-Pro portion of cyclic peptides having the disulfide linkage. Calculated results indicate that four cyclic Ac-Cys-Pro-Xaa-Cys-NHMe essentially form type III beta-bend at the Pro-Xaa portion, and also show fairly good agreement with experimental results of the NMR spectroscopy and X-ray crystallography for the tetrapeptides having Cys-Pro-Xaa-Cys sequence.

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Language：Japanese   Publisher：Japan Thermosetting Plastics Industry Association  

The purpose of this investigation is to improve the toughness, water-proof, and electrical insulation of phenolic resin. For this purpose, distance between two crosslinking nodes of phenolic resin was made longer and density of phenolic OH group was made lower. In the concrete, glutaraldehyde was used instead of formaldehyde. At first, reaction condition such as the kind of solvent or catalyst was investigated in order to synthesize novolac type phenol-glutaraldehyde resin. As a result, the reaction was easily proceeded when acetic acid and HCI were used as solvent and catalysis respectively. Next, flowability and curing behavior of molding compounds which were prepared from phenol novolac, the phenol-glutaraldehyde resin, hexamethylenetetramine as curing agent, and wood flour as filler were investigated. As a result, when the content of the phenol-glutaraldehyde resin was less than 60wt%, the molding compounds were suitable for transfer molding. Curing rate was not depended on the content of the phenol-glutaraldehyde resin. Properties of cured resin by transfer molding from the molding compounds were also investigated. It was found that toughness, water-proof, and electrical insulation were improved by an increase of the content of phenol-glutaraldehyde resin.

DOI： 10.11364/networkpolymer1996.19.87

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Other Link： https://jlc.jst.go.jp/DN/JALC/00053622600?from=CiNii

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：NATURE PUBLISHING GROUP  

Theoretical conformational analysis was carried out on four cyclic tetrapeptides Ac-Cys-Pro-Xaa-Cys-NHMe (Xaa=Val, Phe, Leu, and norleucine) using Empirical Conformation Energy Program for Peptides (ECEPP) and optimization procedure for investigating the effects of differences in the hydrophonbic side-chain groups of Xaa residue on the beta-bend conformation at the Xaa-Pro portion of cyclic peptides having the disulfide linkage. Calculated results indicate that four cyclic Ac-Cys-Pro-Xaa-Cys-NHMe essentially form type III beta-bend at the Pro-Xaa portion, and also show fairly good agreement with experimental results of the NMR spectroscopy and X-ray crystallography for the tetrapeptides having Cys-Pro-Xaa-Cys sequence.

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Language：English   Publishing type：Research paper (international conference proceedings)   Publisher：PROTEIN RESEARCH FOUNDATION  

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Language：English   Publishing type：Research paper (international conference proceedings)   Publisher：FUDAN UNIVERSITY  

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：VSP BV  

Tetrapeptides, Arg-Gly-Asp-SeT (RGDS), Arg-Gly-Asp-Val (RGDV), and Arg-Gly-Asp-Thr (RGDT), respectively, appearing in the cell-attachment domains of fibronectin, vitronectin, and collagen, and pentapeptide Tyr-Ile-Gly-Ser-Arg (YIGSR) appearing in B1 chain of laminin, were synthesized by liquid-phase procedure. Bioactivities of RGD, RGDX (X=S, V and T), YIGSR, and YIGSR-NH2 as cell recognition determinants were investigated by cell-attachment test using these oligopeptides immobilized to ethylene-acrylic acid copolymer (PEA) film. The cell lines used were A431, NRK, CHO-K1, HeLa.S3, and RLC-16 cells. It was found that the residue X in RGDX plays an important role for cell-attachment activity of RGDX, and, regarding YIGSR, introduction of NH2 residue at the C-terminal of the pentapeptide enhances the cell-attachment activity.

Web of Science

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：SOC POLYMER SCIENCE JAPAN  

Pentapeptide Tyr-Ile-Gly-Ser-Arg (YIGSR), an amino acid sequence existing in the cell attachment domain of the B1 chain of laminin, was synthesized by an improved liquid-phase procedure. Bioactivity of YIGSR as cell recognition determinant was investigated using L-929 fibroblast cell and A431 epidermoid cell. The YIGSR pentapeptide was immobilized to ethylene-acrylic acid copolymer (PEA) film, and the number of cells attached to the immobilized film was counted. In addition, the cell-attachment activity of YIGSR toward the cells was also evaluated by the cell inhibition test. These two tests led to a conclusion that YIGSR acts as a cell determinant toward L-929 and A431 cells.

Web of Science

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：JOHN WILEY & SONS INC  

Tetrapeptides, Arg-Gly-Asp-Ser (RGDS), Arg-Gly-Asp-Val (RGDV), and Arg-Gly-Asp-Thr (RGDT), present in the cell-attachment domain of fibronectin, vitronectin, and collagen, respectively, were synthesized by using an improved liquid-phase procedure. Bioactivities of RGD and RGDX (X = S, V, and T) as cell recognition determinants were investigated by two methods to evaluate interactions of these oligopeptides with L-929 fibroblast cells originating in mouse epithelia. In the first method, these oligopeptides were immobilized to ethylene-acrylic acid copolymer (EAA) film, and the cell-attachment activity of the immobilized film was measured. In the second method, interaction of oligopeptides with the cells was evaluated by measuring the cell inhibition caused by oligopeptides. It was found that RGD and RGDX exhibit remarkable cell-attachment activity, and the activity of oligopeptides depends on X.

Web of Science

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：JAPAN SOC BIOSCI BIOTECHN AGROCHEM  

The enzymic peptide synthesis in an organic medium was investigated by using papain in the presence of PEG. The added PEG enhanced the enzyme activity of papain for synthesis of the peptide, Boc-Gly-Asp(OBzl)OBzl. The activity papain in the PEG-added system was higher than that in a PEG-free system.

Web of Science

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：SPRINGER VERLAG  

Tetrapeptide Arg-Gly-Asp-Ser (RGDS), an amino acid sequence existing in the cell-attachment domain of fibronectin, was synthesized using improved solid-phase procedure. Cell-attachment activity of the RGDS toward L-929 fibroblast cells originating in mouse epithelia was examined by measuring (a) the number of cells attached onto RGDS-immobilized polyvinyl alcohol (PVA) film, and (b) the % inhibition of cell-attachment onto polystyrene substrate from suspension of the cells in the presence of RGDS molecules. It was found that (a) a number of cells attached to the RGDS-immobilized PVA films, and (b) RGDS molecules remarkably attached to the cells, and, as a result, RGDS inhibited the cells to adhere onto the substrate.

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：SOC POLYMER SCIENCE JAPAN  

Web of Science

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Language：English   Publishing type：Research paper (scientific journal)  

An oligopeptide, L-arginyl-glycyl-L-aspartyl-L-serine, having cell attachment activity was synthesized from the respective aminoacids carrying suitable protecting residues, by using carboxymethyl polyethylene glycol (PEG)-modified proteases in organic solvents. Papain, trypsin, and a-chymotrypsin were modified with PEG. Organic solvents used were 1, 1, 1-trichloroethane, chloroform and chloroform/ethyl cellosolve (1:1) mixture. Identification of the products was done by gel permeation chromatography (GPC) and thin layer chromatography (TLC). © 1989, Vsp. All rights reserved.

DOI： 10.1163/156856290X00044

Scopus

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Language：Japanese  

J-GLOBAL

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Language：English   Publishing type：Research paper, summary (international conference)   Publisher：MARY ANN LIEBERT, INC  

Web of Science

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J-GLOBAL

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J-GLOBAL

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J-GLOBAL

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Language：Japanese   Publisher：The Japan Society of Mechanical Engineers  

CiNii Books

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Language：Japanese   Publisher：The Japan Society of Mechanical Engineers  

CiNii Books

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Language：Japanese   Publisher：The Japan Society of Mechanical Engineers  

CiNii Books

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J-GLOBAL

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J-GLOBAL

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Language：Japanese   Publisher：The Japan Society of Mechanical Engineers  

Biocompatible piezoelectric materials are becoming increasingly important for actuators and sensors in medical devices. In this paper, we highlighted on some perovskite-type oxides MgSiO_3, MgTiO_3 CaTiO_3, CaZrO_3 and CaSnO_3 which were discovered through first-principles calculation in our previous studies to fabricate novel lead-free biocompatible piezoelectric materials. In order to verify their biocompatibility, the cytotoxicity of isotopic oxides with the same components was examined as comparing with typical perovskite-type oxides BaTiO_3, LiNbO_3, KNbO_3 and NaNbO_3. The fibroblast (L929) cells were cultured during 7 days and the effect of materials was evaluated by the half maximal inhibitory concentration (IC50). As a result, it was recognized that BaTiO_3, MgSiO_3, CaTiO_3 and CaSnO_3 has the higher biocompatibility. On the other hand, the others has lower biocompatibility. Especially LiNbO_3 and NaNbO_3 shows the strong toxicity.

CiNii Books

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Language：English   Publishing type：Research paper, summary (international conference)   Publisher：AMER CHEMICAL SOC  

Web of Science

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Language：Japanese   Publisher：The Japan Society of Mechanical Engineers  

Biocompatible piezoeleciric materials are becoming increasingly important for actuators and sensors in medical devices. In this paper, we highlighted on some perovskite-type oxides MgSiO_3, CaSnO_3, and MgTiO_3 which were discovered through first-principles calculation in our previous studies to fabricate new Irad-free biocompatible piezoelectric materials. In order to verify their biocompatibility, the cytotoxicity of isotopic oxides with the same components was examined as comparing with typical perovskite-type oxides KNbO_3 and LiMbO_3. The fibroblast (L929) cells were cultured during 7 days and the effect of materials was evaluated by the half maximal inhibitory concentration (IC50). As a result, it was recognized that MgSiO_3 and CaSnO_3 has the higher biocompatibility. On the other hand, LiNbO_3 shows the strong toxicity.

DOI： 10.1299/jsmemm.2010.12

CiNii Books

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Language：Japanese   Publisher：[日本材料学会]  

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Language：Japanese  

CiNii Books

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Language：Japanese   Publisher：Kansai University  

CiNii Books

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Other Link： http://hdl.handle.net/10112/947

Language：Japanese   Publisher：The Japan Society of Mechanical Engineers  

本研究では,医用デバイス用アクチュエータやセンサへ適用される生体適合圧電材料として,第一原理計算により求めたペロブスカイト型酸化物であるMgSiO_3, CaSiO_3, CaTiO_3および既存圧電材料であるPb(Zr,Ti)O_3, BaTiO_3に対して細胞毒性実験を行った.マウスの皮下結合組織由来の繊維芽細胞株を各試料の抽出液中において7日間培養し,相対細胞増殖率および倍加時間により評価した.その結果,MgSiO_3およびCaTiO_3が高い生体適合性を示すことを明らかにした.

CiNii Books

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Language：English   Publishing type：Research paper, summary (international conference)   Publisher：AMER CHEMICAL SOC  

Web of Science

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Language：Japanese  

CiNii Books

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Language：Japanese   Publisher：The Japan Society of Mechanical Engineers  

CiNii Books

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Event date： 2018.12

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