| Project/Area Number |
14207055
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| Research Category |
Grant-in-Aid for Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Orthopaedic surgery
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| Research Institution | The University of Tokyo |
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Principal Investigator |
MABUCHI Kunihiko The University of Tokyo, Graduate School of Information Science and Technology, Professor, 大学院・情報理工学系研究科, 教授 (50192349)
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| Co-Investigator(Kenkyū-buntansha) |
NAKAYAMA Yasuhide National Cardiovascular Center, Department of Biomedical Engineering, Laboratory Chief, 生体工学部, 部長 (50250262)
IKEGAWA Shiro RIKEN, SNP Research Center, Laboratory Head, 遺伝子多型研究センター, チームリーダー (30272496)
MATSUDA Takehisa Kushu University, Graduate School of Medicine, Professor, 大学院・医学系研究科, 教授 (60142189)
NAKAMURA Kouzo The University of Tokyo, Graduate School of Medicine, Professor, 大学院・医学系研究科, 教授 (60126133)
HOSHIKAWA Atsuto Dokkyo University School of Medicine, Department of Orthopaedics, Lecturer, 整形外科, 講師 (50383021)
鈴木 隆文 東京大学, 大学院・情報理工学系研究科, 科学技術振興特任教員(常勤形態) (50302659)
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| Project Period (FY) |
2002 – 2004
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| Project Status |
Completed (Fiscal Year 2004)
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| Budget Amount *help |
¥42,770,000 (Direct Cost: ¥32,900,000、Indirect Cost: ¥9,870,000)
Fiscal Year 2004: ¥8,190,000 (Direct Cost: ¥6,300,000、Indirect Cost: ¥1,890,000)
Fiscal Year 2003: ¥13,000,000 (Direct Cost: ¥10,000,000、Indirect Cost: ¥3,000,000)
Fiscal Year 2002: ¥21,580,000 (Direct Cost: ¥16,600,000、Indirect Cost: ¥4,980,000)
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| Keywords | Photo-polymerizing Gelatin / Chondrocyte Transplantation / scaffold / Bone marrow interstitial stem cell / free radical / Type II collagen / aglycan / camphorquinone / Type II コラーゲン / type IIコラーゲン / tvpe II コラーゲン |
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| Research Abstract |
We have developed a styrenated gelatin as a scaffold for cartilage chondrocyte trans-plantation based on the following three-point concept : 1)The scaffold can be cross-linked in situ (by visible light irradiation) at the surgery site to fill any gaps in the articular cartilage tissue ; 2)Transplanted cells enclosed with in it can proliferate and produce cartilage matrices, and ; 3)The scaffold has proper hardness and elasticity and yet will biodegrade after it is cross-linked. In this study, we examined the potential and the problems of styrenated gelatin as a scaffold for chondrocyte transplantation.
Styrenated gelatin was synthesized by connecting styrene groups with lysine gelatin residue and mixed with cultured rabbit chondrocytes in a medium containing 20% fetal bovine serum with camphorquinone equivalent to 0.1% of the gelatin weight. Then the mixed solution was irradiated with harmless visible light (400-520 nm) for 2 minutes using a dental halogen lamp and cross-linked.
The resu
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lts showed that : 1)Embedded chondrocytes can survive, proliferate, and produce cartilage matrices after it is cross-linked under the condition that the substitu-tion ratio of the stylenated group was below ca. 45% (group/molecule), and the mixture weight ratio between the stylenated gelatin and its dissolvent (medium) was 30% ; 2)RT-PCR showed that the articular chondrocytes cultured in the gelatin hydrogel expressed the mRNA of type II collagen and aggrecan core protein ; these cartilaginous matrices were observed histologically throughout the gelatin three weeks after the cell trans-plantation. However, sulfated glycosaminoglycan production, a representative component of the cartilage matrix, was not as high (40-50%) as when collagen gel instead of styrenated gelatin was used as the scaffold ; in this case, the substitution ratio of the stylenated group was ca. 45% group/molecule, and the weight ratio of the mixture between the stylenated gelatin and its dissolvent medium was 30% (the elasticity of the stylenated gelatin hydrogel was around 13-15 KPa (ca. 1/40 of the actual cartilage tissue of the living body) just after it was photo-polymerized).
These results suggest that, even though the embedded chondrocyte proliferation and cartilage matrix production in the stylenated gelatin hydrogel are insufficient and the elasticity and biodegradability of its cross-linked body in-vivo are still unsatisfactory, proving that our chondrocyte transplantation scaffold concept is realizable and promising. Less
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