Figure 2.

Quantitative analysis of osteoclast formation and the pit formation assay in experiment groups and control groups. In experiment groups, MLO-Y4 and RAW264.7 were co-cultured on a plastic dish or dentin slice for eight days, treated with different level of heparin, in the absence or presence of 10 nM 1,25(OH)₂D₃. In control groups, RAW264.7 were cultured with or without MLO-Y4, in the absence or presence of 10 IU/mL heparin and 10 nM 1,25(OH)₂D₃. 50 ng/mL RANKL induced osteoclastogenesis served as a positive control. (A) Osteoclast numbers per well in experiment groups (n = 7); (B) osteoclast numbers per well in control groups (n = 5); (C) area of osteoclastic resorption pits on dentin slices in experiment groups (n = 3); (D) area of osteoclastic resorption pits on dentin slices in control groups (n = 3); (E) giant osteoclasts (osteoclast with more than 20 nuclei) number in experiment group supplemented with 1,25(OH)₂D₃ (n = 5); and (F) giant osteoclast (200×). The black arrows indicate the nuclei in a single giant osteoclast. Each of above values is expressed as the mean ±S.D. MLO: MLO-Y4 cells. RAW: RAW264.7 cells. vitD: 1,25(OH)₂D₃. TRAP: tartrate resistant acid phosphatase. MNC: multinuclear cell. * p < 0.05; ** p < 0.01; *** p < 0.001 represent significant differences from the relevant group 0.