Figure 1.

Effect of PEE on the viability of BMMs and TRAP⁺ MNC formation. Isolated BMMs were treated with PEE in a dose- and time-dependent manner. (A) Cells were treated with various concentrations of PEE (0, 25, 50, 100 and 200 µg/ml) and incubated for 4 days. (B) Cells were treated with 25 µg/ml PEE and incubated for various durations (0, 1, 2, 3 and 4 days). (C) Intracellular TRAP was stained to evaluate the effect of PEE on osteoclastogenesis. TRAP staining of osteoclasts was conducted following 4 days culture in the presence of 0, 5, 10, 25 and 50 µg/ml PEE. Scale bar, 200 µm, at ×10 magnification. (D) Quantification of osteogenesis. TRAP⁺ MNCs with >3 nuclei were classed as mature osteoclasts. (E) TRAP activity was measured with a TRAP solution assay at day 4. Absorbance was measured at 405 nm. Data are presented as the mean ± standard deviation and are representative of at least three experiments. **P<0.05, ***P<0.01 vs. 0 µg PEE. PEE, Peucedanum japonicum Thunb. ethanol extract; BMM, bone marrow-derived macrophage; TRAP, tartrate resistant acid phosphatase; MNC, multinuclear cell.