| Project/Area Number |
13470381
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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 |
Plastic surgery
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| Research Institution | Tokai University |
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Principal Investigator |
TANINO Ryuzaburo Tokai University, School of Medicine, Professor, 医学部, 教授 (50051595)
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| Co-Investigator(Kenkyū-buntansha) |
INOKUCHI Sadaki Tokai University, School of Medicine, Professor, 医学部, 教授 (60160008)
NAKAZAWA Hiroe Tokai University, School of Medicine, Professor, 医学部, 教授 (20110885)
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| Project Period (FY) |
2001 – 2003
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| Project Status |
Completed (Fiscal Year 2003)
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| Budget Amount *help |
¥7,900,000 (Direct Cost: ¥7,900,000)
Fiscal Year 2003: ¥2,400,000 (Direct Cost: ¥2,400,000)
Fiscal Year 2002: ¥2,500,000 (Direct Cost: ¥2,500,000)
Fiscal Year 2001: ¥3,000,000 (Direct Cost: ¥3,000,000)
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| Keywords | cultured skin / vascular inductio / angiogenesis / gene transfer |
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| Research Abstract |
Growth factors such as vascular endothelial growth factor (VEGF) increase in the transplantation floor of cultured skin substitute (CSS) to stimulate angiogenesis. VEGF in the culture supematant in fibrin types of CSS were measured by EHSA, and VEGF mRNA was determined semiquantitatively by RT-PCR after two days of incubation. Furthermore CSS were implanted to athymic mice. VEGF concentration in the fibrin-cultured supematant was 84.3±11.8 pg/mi, whereas it was 27.8±4.68 pg/ml in the ease of the collagen substrate. The relative levels of VEGF mRNA were 1.088±0.100 and 0.698±0.226, respectively. In in vivo-transplantion, the fibrin-type cultured skin substitute showed an excellent "take" on the wound bed, and a normally proliferating keratinocyte layer with emergence of vascular endothelial cells in the transplanted floor was seen three days
To add the ability of angiogenesis to the CSS, gene transfer such as VEGE was planed In order to develop gene induction using virus vector, we chose the Lentivirus. And to examine the effect of gene expression level, Green Fluorescent Protein (GFP) was used as a reporter gene in vitro and used E. coli beta-galactosidase (LacZ) in vivo. Both reporter gene were well transfened to human nonnal Keratinocytes. It conduded lentivirous is effective to introduce the vascular endothelial growth factor to the human kemtinocytes in order to develop the Vascular Inducible Cultured Skin Substitute (VICSS)
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