| Project/Area Number |
09671819
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Ophthalmology
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| Research Institution | TOKYO DENTAL COLLEGE |
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Principal Investigator |
SHIMAZAKI Jun Tokyo Dental College, Department of Dentistry, Lecturer, 歯学部, 講師 (40170930)
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| Co-Investigator(Kenkyū-buntansha) |
SHINOZAKI Naoshi Tokyo Dental College, Department of Dentistry, Lecturer, 歯学部, 講師
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| Project Period (FY) |
1997 – 1998
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| Project Status |
Completed (Fiscal Year 1998)
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| Budget Amount *help |
¥3,000,000 (Direct Cost: ¥3,000,000)
Fiscal Year 1998: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1997: ¥2,200,000 (Direct Cost: ¥2,200,000)
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| Keywords | fibroblasts / conjunctival epithelium / amniotic membrane / growth factors / cell proliferation / Ocular Surface / cicatricial keratoconjunctivitis / dry eye |
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| Research Abstract |
1. Histology and preservation : Fresh human amniotic membrane (AM) had uniform thickness of collagen layer and overlying epithelial cells. The epithelial cells lost their viability up to 80% after 70 days' cryopreservation with DMSO.
2. Animal modesl : Human preserved AMs were secured on bare sciera of albino rabbits and histological changes were studied. The amniotic epithelium remained for about one week postoperatively, then gradually replaced by host conjunctiva. Inflammatory cells were observed beneath the membrane in some cases.
3. Clinical samples : AMs were recognized in 3 out of 14 human samples which had reoperation following AM transplantation, suggesting AMs were not degraded in some cases after transplantation.
4. Growth factors : Roles of growth factors on proliferation/differentiation of ocular surface epithelia were studied using immunohitochemistry and ELISA.bFGF, HGF, and TGF-beta3 were found primarily in the cytoplasm of AM epithelium, whereas TGFbeta 1 and TGF-beta3 were distributed more diffusely. Concentrations of bFGF, HGF, TGF-betal, TGF-beta3 in AM were 5.31*3.04, 14.2*6.0, 2.45*1 .45.0.56*0.45 ng/g, respectively. The concentrations decreased to 1 7.7-23.5% after 70 days' cryopreservation.
5. Effects on ocular surface cells : Addition of homogenized AMs to either cultured conjunctival epithelium or subccnjunctival fibroblasts resulted in significant increases in proliferation of these cells.
6. Cultivation of conjunctival epithelium on AM : We cultivated human conjunctival epithelium on AM using SHEM and RCGM media. These cells became multilayered with intercellular complex formation.
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