Dual Effect of Chrysanthemum indicum Extract to Stimulate Osteoblast Differentiation and Inhibit Osteoclast Formation and Resorption In Vitro

Figure 1

CIE inhibits the formation of TRAP-positive osteoclasts from cocultures and BMMs. (a) Mouse bone marrow cells and primary osteoblasts were cocultured with IL-1 (10 ng/mL) in the presence of CIE (50 μg/mL) or DMSO (control) for 5 days. Cells were fixed with 3.7% formalin in PBS, permeabilized with 0.1% Triton X-100 in PBS, and stained with TRAP solution. (b) Mouse primary osteoblasts were pretreated with CIE (50 μg/mL) or DMSO (control) for 1 h and then were stimulated with IL-1 (10 ng/mL) for indicated times. The expressions of RANKL and OPG were determined by using real-time RT-PCR analysis. (c) BMMs were cultured for 4 days in the presence of M-CSF (30 ng/mL) and RANKL (100 ng/mL) with either DMSO or CIE. Cells were detected by TRAP staining. (d) TRAP-positive multinucleated cells were counted as osteoclasts. ; . (e) BMMs were cultured for 3 days with the indicated doses of CIE in the presence of M-CSF (30 ng/mL). Cell viability was analysed using an XTT assay.