| Project/Area Number |
08672114
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
Functional basic dentistry
|
|---|
| Research Institution | HOKKAIDO UNIVERSITY |
|---|
Principal Investigator |
FUJISAWA Ryuichi HOKKAIDO UNIV., FAC.OF DENTISTRY,INSTRUCTER, 歯学部, 助手 (40190029)
|
|---|
| Project Period (FY) |
1996 – 1997
|
| Project Status |
Completed (Fiscal Year 1997)
|
| Budget Amount *help |
¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 1997: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1996: ¥1,700,000 (Direct Cost: ¥1,700,000)
|
|---|
| Keywords | bone metabolism / cell attachment / bone sialoprotein / hydroxyapatite / 骨 / RGDペプチド / インテグリン |
|---|
| Research Abstract |
We synthesized a peptide which mimics functional sequences of bone sialoprotein (BSP), a cell attachment protein in bone. This BSP model peptide (E_7PRGDY) is composed of a hydroxyapatite-binding sequence (E_7) and a cell-attachment sequence (RGD) of BSP.This peptide has affinity to synthetic hybroxyapatite crystals.
We examined attachment osteoblastic MC3T3-E1 cells to the crystals coated with proteins. The cells were attached to the crystals coated with BSP or the peptide, although they were not attached to the crystals coated with serum albumin. The cells attached on the peptide-coated crystals were flattened and developed pseudopods. The attachment was inhibited with excessive amount of a RGD peptide. The peptide may be attached to the crystals through the poly (Giu) sequence and expose the RGD sequence to a medium. Integrins on the surface of the cells can interact with RGD sequence.
We also examined attachment of osteoclastic cells to the crystals coated with proteins. The osteoclastic cells were prepared by differentiating murine bone marrow cells. The bone marrow cells were cultured on the crystals in a medium containing 1,25 (OH) _2D_3. Osteoclastic cells were detected by staining for tartrate-resistant acid phosphatase. Osteoclastic cells were observed attached on the crystals coated with BSP or the BSP model peptide. The attachment was inhibited with excessive amount of the RGD peptide, indicating involvement of integrins in this attachment. We conclude that BSP can mediates attachment of osteoblasts and osteoclasts on bone mineral and that its poly (Glu) sequence and RGD sequence are involved in this function.
|
