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Hybrid Scaffolds Composed of Amino-Acid Coated Sponge and Hydroxyapatite for Hard Tissue Formation by Bone Marrow Cells

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Journal of Biomedical Science and Engineering 2014年第6期7卷 316-329页

作者： Takayoshi Yabuuchi Masataka Yoshikawa hideyuki kakigi Hiroyuki Hayashi Department of Endodontics Osaka Dental University Osaka City Japan

A formalin-treated polyvinyl-alcohol (PVF) sponge is convenient as a scaffold because its configuration is easily modified. However, coating the sponge with an adhesive chemical agent is necessary to attach bone marrow cells (BMCs) to the sponge structure. Moreover, it was considered that a hybrid scaffold composed of a sponge and enveloped cylindrical porous hydroxyapatite (HA) would be convenient. In this study, the effect of leucine (Leu) coating on a PVF sponge was examined for osteogenesis on an HA/PVF hybrid scaffold by rat BMCs (rBMCs). In an in vivo assessment, the sponge immersed in Leu solution (10 mg/ml) was inserted into the hollow center of cylindrical HA. The sponge received 1.5 × 106 rBMCs obtained from male Fischer 344 rats. The hybrid scaffolds were then implanted subcutaneously of syngeneic rats for 6 weeks. In vitro assessment of Leu to hard tissue formation with coating on the well or addition in rBMC culture medium was also performed in a 6-well plate for 2 weeks. In vivo examinations showed the excellent effect of Leu coating on PVF sponge. Leu-coated PVF sponge in the scaffolds showed marked new bone formation in the pores by histological examination. Leu-coated PVF sponge showed a high quantity of osteocalcine (OC). HA might prevent the release of rBMCs from PVF as a barrier. In in vitro examinations, the quantity of OC in rBMC culture with and without the addition of Leu in culture medium showed no significant difference. However, addition of Leu showed significant ALP activity level in culture medium. Leu coating in culture plate wells showed no influence on the quantity of OC. It was concluded from the results that Leu might prevent the emigration of rBMCs to the outside of the scaffold and promote the differentiation of cells to osteoblasts in the scaffold.

关键词： Amino-Acid Bone Marrow Cells Bone Formation Hybrid Scaffold Scaffold Hydroxyapatite Formalin-Treated Polyvinyl-Alcohol Sponge

Effect of L-lysine in culture medium on nodule formation by bone marrow cells

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Journal of Biomedical Science and Engineering 2012年第10期5卷 587-592页

作者： Masataka Yoshikawa Yasunori Shimomura, hideyuki kakigi Norimasa Tsuji Takayoshi Yabuuchi Hiroyuki Hayashi Department of Endodontics Osaka Dental University Osaka Japan

The aim of this study was to estimate the effect of L-lysine on nodule formation by rat bone marrow cells in vitro. In this study, L-lysine was added to medium for mesenchymal stem cell culture to promote proliferation and differentiation of the cells, and then nodule formation was estimated in an in vitro rat bone marrow cell culture. Bone marrow cells from the bone shafts of the femora of Fischer 344 rats were cultured in minimum essential medium with 20 μl of L-lysine solution at 10﹣4, 10﹣5, 10﹣6, 10﹣7 or 10﹣8 M. Dexamethasone was also added to the medium at 10 nM for differentiation of stem cells from bone marrow into osteoblast progenitor cells. The subculture was performed for 2 weeks. The quantity of osteocalcin in rat bone marrow cell culture with dexamethasone was 392 ng/ml. In the medium with dexamethasone and 10﹣8 M L-lysine, the quantity of osteocalcin was 437 ng/ml. Nodules only formed upon addition of 20 μl of L-lysine at 10﹣8 M. It was indicated that 10﹣8 M L-lysine should be the optimal concentration for calcification. For nodule formation by rat bone marrow cells in vitro, the optimum concentration of L-lysine in culture medium might be 20 μl of 10﹣8 M. L-lysine could play an important role in matrix production for bone formation in vitro.

关键词： L-Lysine Bone marrow Cells Calcium Deposition Dexamethasone In Vitro

Dextran coating on and among fibers of polymer sponge scaffold for osteogenesis by bone marrow cells in vivo

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Journal of Biomedical Science and Engineering 2010年第8期3卷 751-757页

作者： Masataka Yoshikawa Norimasa Tsuji hideyuki kakigi Takayoshi Yabuuchi Yasunori Shimomura Hiroyuki Hayashi Hajime Ohgushi 不详

Although hydroxyapatite is commonly used as a scaffold for bone regeneration, sponges may be suitable because of the adaptability to the defect. To use as a scaffold, the fiber of sponge would be coated with any adhesive to storage stem cells in the sponges. Fiber in the structure of commercially available sponges was coated by immersion in dextran solution and air dried. After seeding of rat bone marrow cells (rBMCs), the sponges were implanted subcutis of rats for estimate osteogenesis in vivo. The level of osteocalcin was 25.28 ±5.71 ng/scaffold and that of Ca was 129.20 ±19.69 μg/scaffold. These values were significantly high- er than those in sponges without dextran coating (p <0.01). It was thought that rBMCs could be stored on the shelf by dextran deposition in the fiber of the sponge. In vivo examination, dextran induced osteogenesis by rBMCs in many spaces in the inner structure of the sponge.

关键词： Dextran Cell Adhesion Scaffold Bone Marrow Cells Osteogenesis

Bone Formation in a Scaffold Composed of Cylindrical Hydroxyapatite and Tryptophan- or Lysine-Coated Sponge in Vivo

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Journal of Biomedical Science and Engineering 2015年第6期8卷 389-398页

作者： Masataka Yoshikawa hideyuki kakigi Hiroshi Maeda Ikuo Nishikawa Hideaki Ikenaga Takeshi Inamoto Norimasa Tsuji Department of Endodontics Osaka Dental University Osaka Japan

Because of the three-dimensional structure of bone or hard tissue such as a tooth, a scaffold is necessary for its regeneration by cellular engineering. Commonly, for in vivo examination, hydroxyapatite (HA) has been used as such a scaffold. Cylindrical HA with a hollow center, which included a columnar formalin-treated polyvinyl alcohol sponge, was used in this examination as a scaffold. The sponge had been coated with L-tryptophan or L-lysine before insertion into the hollow center of the HA. Rat bone marrow cells (rBMCs) derived from the femur were seeded in the sponge before insertion into the hollow center of HA. The number of rBMCs seeded in each sponge was 1.5 × 106. These scaffolds were implanted subcutaneously into the backs of Fischer 344 rats for 6 weeks. In the amino-acid-coated sponge in HA, osteogenesis was found histologically. An osteocalcin level of approximately 10 ug was measured in the scaffolds with L-tryptophan-coated formalized poly-vinyl alcohol sponge containing rBMCs, 4 ug on average in the scaffolds with L-lysine-coated sponge containing the cells and about 2 ug in each scaffold with uncoated sponge containing the cells. The structure of the scaffolds used in this study was thought to be suitable for osteogenesis by rBMCs. It was concluded that tryptophan, as a factor for bone formation by stem cells, functioned by promoting cell adhesion and the differentiation of stem cells into osteoblasts.

关键词： Scaffold Hydroxyapatite Bone Marrow Cells L-Tryptophan L-Lysine Poly-Vinyl Formal Sponge

In Vivo Estimation of Osteogenesis by Bone Marrow Cells in a Bi-Phasic Scaffold and in Each of Its Components

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Journal of Biomedical Science and Engineering 2016年第11期9卷 501-514页

作者： Masataka Yoshikawa hideyuki kakigi Ayano Miyamoto Sadaomi Sugimoto Keisuke Nakai Hideaki Ikenaga Takeshi Inamoto Hiroshi Maeda Department of Endodontics Osaka Dental University Osaka Japan

A bi-phasic scaffold consisting of a columnar formaldehyde-acetalized polyvinyl alcohol (PVF) sponge and a cylindrical porous hydroxyapatite (HA) with a hollow center was devised. Rat bone marrow cells (rBMCs) were seeded into the sponge placed in the hollow center of the cylindrical porous HA. The bi-phasic scaffold, a cylindrical porous HA and a PVF sponge separated from a bi-phasic scaffold after rBMC seeding, and a PVF sponge without rBMCs as a negative control, were implanted for 6 weeks into rat dorsal subcutaneous tissue. In each construct, bone formation was examined histologically and osteocalcin was measured immunochemically. Bone formation was observed in the bi-phasic scaffold and also in the cylindrical porous HA isolated from the bi-phasic scaffold. A significant difference in the quantity of osteocalcin was observed between the bi-phasic scaffold and the isolated cylindrical porous HA. No bone formation was found in the isolated PVF sponge. The bi-phasic scaffold as an outer layer of the scaffold seemed to inhibit the outflow of rBMCs from the PVF sponge. This type of bi-phasic scaffold may have two specific characteristics: Attachment of cells both in PVF sponge and cylindrical porous HA.

关键词： Bi-Phasic Scaffold Hydroxyapatite Polyvinyl Alcohol Sponge Bone Formation In Vivo

Direction without speed information process in the human brain: a magnetoencephalographic study using random dots apparent motion stimulus

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Open Journal of Molecular and Integrative Physiology 2011年第2期1卷 17-22页

作者： Sachiko Oka Tomokazu Urakawa Ryusuke kakigi Yoshiki Kaneoke Department of Integrative Physiology National Institute for Physiological Sciences Myodaiji-cho Okazaki Japan

Apparent motion stimulus induces visual perception of smooth motion even though there is no speed information. We examined whether human brain response as measured by magnetoencephalography carries direction information in the visually presented apparent motion of a randomdot pattern in a similar manner as continuous motions that have speed and direction information. Although there was no significant effect of motion direction on the peak response latency and amplitude, mutual information entropy (IE) significantly increased after the motion onset at approximately 36 ms after the response latency in 41% of the evaluations. Detailed analysis of the data from five subjects who participated in both the present apparent motion and our previous coherent motion studies revealed that the maximum IE latency (delay) for apparent motion was significantly longer than that for coherent motion, although the mean maximum IE was the same. The results indicate that direction is represented in the response waveform evoked by apparent motion but the manner is different from that for coherent motion probably due to the distinct neural processes engaged only for the apparent motion perception. We consider that direction and speed can be processed separately in the human brain because direction information was generated without speed information for the perception of apparent motion.

关键词： Apparent Motion Random Dot Kinematogram Direction Information Entropy Magnetoencephalography

虾青素摄入、热应激和间歇式再负荷的联合处理对废用性肌萎缩的肌卫星细胞的影响（英文）

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Journal of Zhejiang University-Science B(Biomedicine & Biotechnology) 2018年第11期19卷 844-852页

作者： Toshinori YOSHIHARA Takao SUGIURA Nobuyuki MIYAJI Yuki YAMAMOTO Tsubasa SHIBAGUCHI Ryo kakigi Hisashi NAITO Katsumasa GOTO Daijiro OHMORI Toshitada YOSHIOKA Graduate School of Health and Sports Science Juntendo UniversityInzaiChiba 270-1695Japan Faculty of Education Yamaguchi UniversityYamaguchiYamaguchi 753-8513Japan Toyo Koso Kagaku Co.Ltd. UrayasuChiba 279-0041Japan Institute of Health and Sports Science Kurume UniversityKurumeFukuoka 839-8502Japan Institute of Liberal Arts and Science Kanazawa UniversityKanazawaIshikawa 920-1192Japan Faculty of Medicine Juntendo UniversityBunkyo-kuTokyo 113-0033Japan Graduate School of Health Sciences Toyohashi SOZO UniversityToyohashiAichi 440-8511Japan Department of Chemistry School of MedicineJuntendo UniversityInzaiChiba 270-1695Japan Hirosaki Gakuin University HirosakiAomori 036-8577Japan

目的:探讨透过虾青素摄入、热应激和间歇式再负荷的不同处理方式及其联合处理,对于尾部悬吊后大鼠比目鱼肌卫星细胞的影响。创新点:利用虾青素摄入、热应激与间歇式再负荷的不同方式,探讨预防大鼠比目鱼肌废用性肌萎缩的有效处理。方法:... 详细信息

目的:探讨透过虾青素摄入、热应激和间歇式再负荷的不同处理方式及其联合处理,对于尾部悬吊后大鼠比目鱼肌卫星细胞的影响。创新点:利用虾青素摄入、热应激与间歇式再负荷的不同方式,探讨预防大鼠比目鱼肌废用性肌萎缩的有效处理。方法:将49只雄性Wistar大鼠分成对照组、吊尾组、间歇式再负荷组、虾青素摄入组、热应激组和虾青素摄入+热应激组。摘取7天尾部悬吊后的大鼠的比目鱼肌,使用免疫组织化学技术,分析肌卫星细胞的激活程度。结论:虾青素摄入、热应激与间歇式再负荷的不同方式的联合处理能减少比目鱼肌废用性肌萎缩,其部分效果可能是由于肌卫星细胞数量的增加所致。

关键词：虾青素废用性肌萎缩热应激肌卫星细胞

Study on Styrene Grafting onto Per-fluorinated Polymers Effects of Irradiated and Reaction Conditions

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Study on Styrene Grafting onto Per-fluorinated Polymers Effe...

Study on styrene grafting onto per-fluorinated polymers——effects of irradiation and reaction conditions

第6届辐射研究与辐射工艺学术年会

作者： Akio Matsuura Tomoyuki kakigi Saneto Asano Fumihiro Mutou Akihiro Oshima Masakazu Washio Advanced Research Institute for Science and Engineering Waseda University 3-4-1 Okubo Shinjuku Tokyo 169-8555 JAPAN

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辐射研究与辐射工艺学报 2005年第2期23卷 103-103页

作者： Akio Matsuura Tomoyuki kakigi Saneto Asano Fumihiro Mutou Akihiro Oshima Masakazu Washio Advanced Research Institute for Science and Engineering Waseda University Shinjuku Tokyo 169-8555 JapanAdvanced Research Institute for Science and Engineering Waseda University Shinjuku Tokyo 169-8555 JapanAdvanced Research Institute for Science and Engineering Waseda University Shinjuku Tokyo 169-8555 JapanAdvanced Research Institute for Science and Engineering Waseda University Shinjuku Tokyo 169-8555 JapanAdvanced Research Institute for Science and Engineering Waseda University Shinjuku Tokyo 169-8555 JapanAdvanced Research Institute for Science and Engineering Waseda University Shinjuku Tokyo 169-8555 Japan

Fuel cell is in focus as a very low exhaust new dynamo system. Especially polymer electrolyte fuel cells (PEFC) using proton exchange membranes (PEM) have attracted much attention for the electrical vehicle and other mobile applications such as cellar phone and personal computer. Conventional perfluoro-sulfonic acid membranes such as Nation() (DuPont de Nemours LTD.) have been the popular PEM used in PFEC. However, there are still several problems such as insufficient gas barrier properties, low thermal resistance, and their high *** this study, hawse have studied the partially fluorinated PEM for PEFC by radiation grafting onto per-fluorinated polymers such as PFA, FEP and ETFE with reactive styrene monomers using pre-irradiation grafting method in gas and in liquid phase, respectively, and successive sulfonation. Characteristic properties of the grafted and sulfonated materials have been discussed and optimized fabricating conditions were *** properties of the obtained materials have been measured by differential scanning calorimetry,thermogravimetric analysis and FT-IR spectroscopy, *** the case of irradiation in air, glass transition temperatures (Tg) of the grafted materials depend on the yields of styrene grafting onto the materials. That is higher yields of grafting give lower thermal stability. On the other hand, when the peroxy radicals were converted trapped radicals by exposure in air after irradiation under oxygen-free atmosphere, Tg of the grafted materials hardly depends on the grafting ***, the yields of grafting in gas phase were higher than that of liquid phase. However, the sample form of grafting in gas phase has been greatly changed, whereas the sample form of grafting in liquid phase has kept its original ***, it is suggested that the fabricating conditions for PEFC membranes were preferred to irradiation under oxygen-free atmosphere and grafting in liquid *** exchange capacities of t

关键词： polymers effects and