Effects of AA and DHA on osteoclast formation and resorption pit formation.

<p>CD14+ monocytes were seeded in 96-well plates and osteoclast differentiation was stimulated by the addition of 25 ng ml^-1 M-CSF and 30 ng ml^-1 RANKL as described in Materials and Methods. Vehicle (0.08% ethanol) or LCPUFAs were added from day 3 for in differentiating osteoclasts and from the onset of resorption (day 12–14) in mature osteoclasts. Experiments were terminated after 3 weeks. Osteoclasts were TRAP stained and counted under a light microscope. Haematoxylin was used to visualize the nuclei as well as the resorption pits on dentine discs. TRAP+ cells with 3 or more nuclei were counted. A. Photomicrographs of the effects of AA and DHA on osteoclast formation in differentiating osteoclasts seeded on glass coverslips. Large multinucleated osteoclasts are indicated with black arrows. Scale bar = 500 μm. B. Effects of AA and DHA on osteoclast formation in differentiating osteoclasts. C. Effects of AA and DHA on mature osteoclast numbers. D. Effects of AA and DHA on resorption pit formation on dentine discs in mature osteoclasts. The results are representative of three independent experiments conducted in triplicate. Diff Ocl—differentiating osteoclasts. Mat Ocl—mature osteoclasts. N.D.—No osteoclasts detected. *P<0.05; **P<0.01; ***P<0.001 vs. vehicle.