Effects of prostaglandin E2 and lipopolysaccharide on osteoclastogenesis in RAW 264.7 cells.
BACKGROUND
Prostaglandins (PGs) can act on both hematopoietic and osteoblastic lineages to enhance osteoclast formation.


METHODS
We examined PGE2 stimulated osteoclastogenesis in RAW 264.7 cells and the role of endogenous PGE2 in lipopolysaccharide (LPS) stimulated osteoclastogenesis.


RESULTS
RANKL (1-100 ng/ml) increased formation of osteoclasts, defined as tartrate resistant acid phosphatase multinucleated cells, with peak effects at 30 ng/ml. Addition of PGE2 (0.01-1.0 microM) to RANKL (30 ng/ml) dose dependently increased osteoclast number 30-150%. Use of NS-398 (0.1 microM) or indomethacin (Indo, 1.0 micro M) to block endogenous PG synthesis had little effect on the response to RANKL alone but significantly decreased the response to PGE2. Addition of LPS (100 ng/ml) to RANKL increased osteoclast number 50%, and this response was significantly decreased by NS-398 and Indo. RANKL and PGE2 produced small, additive increases in COX-2 mRNA levels, while LPS produced a larger increase. PG release into the medium was not increased by RANKL and PGE2 but markedly increased by LPS.


CONCLUSIONS
We conclude that RANKL stimulated osteoclastogenesis can be enhanced by PGE2 and LPS though direct effects on the hematopoietic cell lineage and that these effects may be mediated in part by induction of COX-2 and enhanced intracellular PG production.
Kaneko
H
H
Musculoskeletal Institute, University of Connecticut Health Center, 263 Farmington Avenue, Farmington, CT 06030, USA.
Mehrotra
M
M
Alander
C
C
Lerner
U
U
Pilbeam
C
C
Raisz
L
L
D000818:Animals; D045744:Cell Line, Tumor; D015232:Dinoprostone / Q000494:pharmacology*; D008070:Lipopolysaccharides / Q000494:pharmacology*; D051379:Mice; D009578:Nitrobenzenes / Q000494:pharmacology; D010010:Osteoclasts / Q000187:drug effects*; D011461:Prostaglandins F, Synthetic / Q000494:pharmacology; D053245:RANK Ligand / Q000494:pharmacology; D013449:Sulfonamides / Q000494:pharmacology
D008070:Lipopolysaccharides; D009578:Nitrobenzenes; D011461:Prostaglandins F, Synthetic; D053245:RANK Ligand; D013449:Sulfonamides; C505689:Tnfsf11 protein, mouse; C080955:N-(2-cyclohexyloxy-4-nitrophenyl)methanesulfonamide; C006326:fluprostenol; D015232:Dinoprostone