1997 Fiscal Year Final Research Report Summary
Dye assay of matrix components in gingival crevicular fluid for destructive activity of alveolar bone
| Project/Area Number |
08457514
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (B)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
Conservative dentistry
|
|---|
| Research Institution | Asahi University |
|---|
Principal Investigator |
IWAYAMA Yukio Asahi Univ.School of Dentistry, Prof., 歯学部, 教授 (70028744)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
KOU Yoshitada School of Dentistry Asahi University Instructor, 歯学部, 助手 (40298435)
SHIBUTANI Toshiaki School of Dentistry, Asahi University Ass.Prof., 歯学部, 講師 (40206149)
|
|---|
| Project Period (FY) |
1996 – 1997
|
| Keywords | matrix / alveolar bone / sulfated glycosaminoglycan / alcian blur / disease activity / gingival crevicular fluid |
|---|
| Research Abstract |
A modification of the method of detecting sulfated glycosaminoglycans (S-GAG) was adapted for use in an Alcian blue assay. We used low pH,and high salt concentration to minimize interference from other polyanions such as DNA,protein, or hyaluronate. The samples of S-GAGs including chondroitin 4-sulfate, chondroitin 6-sulfate, dermatan sulfate and heparan sulfate were blotted onto cellulose acetate strips and stained with 0.2% Alcian blue solution containing 0.05M MgCl^2,0.4M guanidine-HCl, 0.002M sulfuric acid, and 0.25% Triton X-100 at pH 1.5. There was some variability in the binding ability of S-GAG species to the dye. Chondroitin 4-sulfate showed the greatest affinity for the dye while heparan sulfate had the weakest.
The amount of dye bound by DNA,bovine serum albumin, or hyaluronate was negligible. The new method developed here can be adapted as a quantitative assay using a scanning densitometer with a linear range to 300ng of S-GAG.Moreover, this assay method can be used for direct measurement of S-GAG in the human biological fluid (saliva, serum, synovial fluid) without interferences by other macromolecular polyanion.
The total S-GAG obtained from the timed samples of gingival crevicular fluid was significantly higher (P<0.001) in the periodontitis subjects (71.41(]SY.+-。[)20.04ng) compared to the healthy (7.83(]SY.+-。[)2.21ng), and gingivitis subjects (10.53(]SY.+-。[)4.28ng).
|
