Fig. 2.

Osteoclast suppression by anti-RANKL mAb in vitro. Primary macrophages were expanded in bone marrow suspension cells by M-CSF treatment. Macrophages were treated with 0, 1, or 10 μg/mL of the anti-RANKL mAb for 3 days and cell numbers (a) were counted. Osteoclastogenesis was performed by stimulating macrophages with M-CSF and RANKL. Osteoclast cultures at day 5 were treated with the anti-RANKL mAb for 24 h and stained for TRAP expression. Average osteoclast numbers (b) and TRAP(+) macrophages (c) were plotted. Primary gingival fibroblasts were isolated from small pieces of gingiva and expanded. Gingival fibroblasts were cultured for 6 days with the mAb treatment every 2 days. Cells were enumerated and plotted (d). All assays were triplicated. *p < 0.05, **p < 0.01, ***p < 0.001