1988 Fiscal Year Final Research Report Summary
Purification of a growth factor from human bone matrix and its clinical application.
| Project/Area Number |
61570708
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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 | Hokkaido University |
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Principal Investigator |
MASUDA Takeshi Hokkaido University School of Medicine, 医学部, 助教授 (20109424)
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| Project Period (FY) |
1986 – 1988
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| Keywords | Bone matrix / Non-collagenous protein / Purification of growth factor in bone / Insulin-like growth factor II / コラーゲン合成の促進 |
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| Research Abstract |
Bone volume is determined by the relative rates of bone resorption and bone formation. The purpose of this study was to purify a growth factor from human bone matrix activating the proliferation of embryonic chick calvarial cells.
Materials and Methods
Cancellous bone matrix from human femoral heads were pulverized, washed to remove serum and debris, and then demineralized in 10% EDTA (pH 7.2). The growth was purified from the EDTA extract by the procedures including acetone treatment, Hw 50F colomn chromatography, sephadex G-75 superfine column chromatography, MONO-Q fast protein liquid chromatography, and reversed-phase high pressure liquid chromatorgaphy.
Results
1 In the final purification step, the growth factor was eluted as a single peak at 114 min. by the reversed-phase HPLC. The protein migrated as a single band upon SDS-PAGE.
2) Sequence of the first 30 N-terminal amino acids of the final preparation were identical to that of Insulin-like growth factor II (IGF-II).
3) The purified growth factor at 0.1 ng/ml doubled DNA synthesis, collagen synthesis and alkaline phosphatase activity compared to the control value determined in the absence of the factor.
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