| Project/Area Number |
62570681
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Orthopaedic surgery
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| Research Institution | Kyoto University |
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Principal Investigator |
SHIMIZU Katsuji Faculty of Medicine, Kyoto University, 医学部, 助手 (90170969)
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| Co-Investigator(Kenkyū-buntansha) |
IWASAKI Renpei Faculty of Medicine, Kyoto University, 医学部, 助手 (70184877)
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| Project Period (FY) |
1987 – 1988
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| Project Status |
Completed (Fiscal Year 1988)
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| Budget Amount *help |
¥2,300,000 (Direct Cost: ¥2,300,000)
Fiscal Year 1988: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1987: ¥1,800,000 (Direct Cost: ¥1,800,000)
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| Keywords | Intervertebral Discs / Ageing / アミロイド |
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| Research Abstract |
Age-associated amyloid deposition in human articular structure has been reported in synovial joints (Uchino et al., 1980; Goffin et al.,1981) and in intervertebral discs (Takeda et al., 1984) by histological studies. While amyloid from various tissues are characterized and classified by the nature of amyloid fibril proteins, such biochemical method of investigation has not been applied to the study of amyloid of articular tissues.
To investigate amyloid of ageing in human interverterbral discs biochemically, validity of the water-solubilization technique by Pras et al. was assessed in the present study.
Sixty surgical specimen of intervertebral discs consisting of annulus fibrosus and nucleus pulposus were collected and stored at -20 C. Thirty intervertebral discs were selected for biochimical studies by their age and by the conditions after preservation.
After pulverization in liquid introgen, tweleve times of extraction of the sample by 150 mM saline washed out most of the soluble proteins. Successive solubilization by distilled water was monitored by absorption at 280 nm. and the first two extracts were collected. Aliquots were processed for electronmicroscopic study after negative staining.
Electron microscopic studies of water solubilized fraction showed a few cluster of amyloid fibrils in about 100 observatory fields. Therefore, further purification or characterization was not attempted.
It was concluded that water-solubilization of amyloid in intervertebral disc yields very small amount of amyloid fibrils and that a low content of amyloid in the tissue or some nuknown effect of the matrix resulted in poor extraction.
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