OHSHIMA Mitsuhiro NIHON UNIVERSITY SCHOOL OF DENTISTRY,ASSISTANT, 歯学部, 助手 (30194145)
TSURUMACHI Tamotsu NIHON UNIVERSITY SCHOOL OF DENTISTRY,LECTURER, 歯学部, 講師 (60139201)
MAENO Masao NIHON UNIVERSITY SCHOOL OF DENTISTRY,ASSOCIATE PROFESSOR, 歯学部, 助教授 (60147618)
TAKEICHI Osamu NIHON UNIVERSITY SCHOOL OF DENTISTRY,ASSISTANT, 歯学部, 助手 (10277460)
|Budget Amount *help
¥2,300,000 (Direct Cost : ¥2,300,000)
Fiscal Year 1999 : ¥900,000 (Direct Cost : ¥900,000)
Fiscal Year 1998 : ¥1,400,000 (Direct Cost : ¥1,400,000)
1. Prostaglandins (PGEィイD21ィエD2, PGEィイD22ィエD2, TXAィイD22ィエD2) stimulated IL-6, TGF-β1 and PDGF-A gene expression and the synthesis of these proteins by radicular cyst-derived fibroblast-like cells (RC cells).
2. PGEィイD22ィエD2 secretion by RC cells were increased 5- to 15-fold when stimulated with IL-1α, -1β, PDGF-AA, -BB, EGF or bFGF. Furthermore, IL-1α-stimulated PGEィイD22ィエD2 production was significantly inhibited by H7 (PKC inhibitor) or dexamethasone.
3. PGEィイD22ィエD2 inhibited DNA synthesis by RC cells, whereas low concentration of TXAィイD22ィエD2 stimulated DNA systhesis. PGEィイD22ィエD2 or TXAィイD22ィエD2 somewhat inhibited collagenase production. TXAィイD22ィエD2 significantly stimulated IL-6 production, and PGEィイD22ィエD2 markedly stimulated LIF synthesis. Both prostanoids stimulated IL-8 synthesis dose-dependently, and marked stimulated MCP-1 production.
4. mRNA levels of cytokines/growth factors in five radicular cyst lining epithelia were analyzed by RT-PCR. The cyst lining epithelia apparently
expressed IL-1α, -1β, IL-6, IL-8, IL-11, TGF-β1, PDGF-A, aFGF, and receptors for KGF, EGF, HGF (c-met) and IL-6.
5. Effects of enzyme digestion on osmotic pressure of various protein solution (albumin, gelatin or transferrin) or hyaluronic acid (HA) solution was determined. Although protein concentrations had little effect on the increase in osmotic pressure, enzyme digestion rate of each protein was significantly effective to increase osmotic pressure (approx. 8-20 mOSM). Countrary to protein solution, low concentration of HA solution (0.1%) revealed approx. 10 mOSM increase in osmotic pressure, whreas the same HA solution increased only 3 mOSM after the enzyme digestion.
6. Active and latent collagenase activities were found in all the cyst fluid specimens, and active collagenase was cross-reacted with anti-human MMP-8 antibody at 65 kDa. Gelatiase activity was also found in all the specimens, which detected as major doublet bands colse to 92 kDa, and a minor band at 72 kDa by zymography ; those were cross-reacted with anti-human MMP-9 and -2 antibody, respectively. All the enzyme activities were blocked by the addition of an MMP inhibitor, o-phenanthroline. Less