| Project/Area Number |
17390382
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Thoracic surgery
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| Research Institution | The University of Tokushima |
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Principal Investigator |
KITAGAWA Tetsuya The University of Tokushima, Institute of Health Biosciences, Graduate School, Professor (80240886)
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| Co-Investigator(Kenkyū-buntansha) |
YOSHIZUMI Masanori Nara Medical University, Faculty of Medicine, Professor (60294667)
KITAICHI Takashi The University of Tokushima, Institute of Health Bioscienoes, Graduate School, Lecturer (20335813)
KANBARA Tamotsu The University of Tokushima, Institute of Health Bioscienoes, Graduate School, Assistant Professor (20380082)
YOSHIDA Homare The University of Tokushima, University Medical and Dental Hospital, Assistant Professor (50448338)
URATA Masahisa The University of Tokushima, University Medical and Dental Hospital, Clinical Fellow (40448337)
増田 裕 徳島大学, 大学院ヘルスパイオサイエンス研究部, 講師 (20314875)
来島 敦史 徳島大学, 医学部・歯学部附属病院, 医員 (50403704)
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| Project Period (FY) |
2005 – 2007
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| Project Status |
Completed (Fiscal Year 2007)
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| Budget Amount *help |
¥11,060,000 (Direct Cost: ¥10,100,000、Indirect Cost: ¥960,000)
Fiscal Year 2007: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000)
Fiscal Year 2006: ¥2,300,000 (Direct Cost: ¥2,300,000)
Fiscal Year 2005: ¥4,600,000 (Direct Cost: ¥4,600,000)
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| Keywords | decellularization technique / scaffold / endothelialization / recellularization in vivo / recellularization in vitro / 滅菌法 / キャホールド / α-SMA / マトリックス / 脱細胞化心臓弁 / 再細胞化 / トリプシン / トライトン / 水酸化アンモニウム / フリーズドライ |
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| Research Abstract |
Background and Objectives : Although various technologies to recellularize both allogenic/xenogenic decellularrized heart valves and tissue-engineered biodegradable scaffolds have been developed, adequate condition of the decellularized matrix scaffold for recellularization by recipient cells and mechanism of regeneration of these grafts remain to be elucidated. The purpose of this study was to find optimal decellularizing techniques for the heart valve from the point of the matrix structure and to evaluate the recellularization by recipient cells using piglet model in vivo.
Adequate decellularization techniques= Newly developed ammonium hydroxide-triton method, even the 0.1% concentration of ammonium hydroxide, decellularized the heart valve tissue completely. Although the elastin and collagen fiber, which is the main matrix structure, was maintained with good condition by the ammonium hydroxide-triton method, most of glycosaminoglycan fiber has almost disappeared. Concerning on the di
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sappearance of glycosaminoglycan, further examination to preserve the glycosaminoglycan fiber will be required for better decellularization.
Recellularization of the decellularized pulmonary valve in vivo : Porcine decellularized monocusp valve graft was implanted to the internal jugular vein of allogenic piglet model, and was harvested four weeks after implantation. Immunohistological examination showed that α-SMA positive cells migrated into the surface and inner layer of the graft tissue, and vWF positive cells covered the surface of it. Matrix structure of the graft seemed to be preserved, however, massive inflammatory cells infiltrated into the graft tissue.
Discussion : We developed 0.1% ammonium hydroxide-triton method as an adequate decellularization technique from the point of matrix tissue preservation. These data on recellularization of the decellularized pulmonary valve implanted in vivo imply that recipient cells migrate into the graft with decellularized matrix scaffold. Concerning on reasonable recellularization of the graft, further examination to exclude the inflammatory cells infiltration and to function with good hemodynamics and durability will be required for better decellularization technology in future clinical use. Less
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