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. 2021 Nov 23;15:4741–4757. doi: 10.2147/DDDT.S334421

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© 2021 Wu et al.

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Saikosaponin D attenuated the formation of osteoclasts induced by RANKL. (A) Chemical structure of SSD. (B) Cell viability of BMMs (bone marrow-derived macrophages) was evaluated by CCK-8 at 48, 96h (OD 450nm). (C) The role of SSD in osteoclast formation in a dose-dependent manner. BMMs stimulated by RANKL (50ng/mL) were treated with or without various concentrations of SSD for 7 days. After fixing, the cells subjected to TRAP staining. (D) The number and area of TRAP-positive multinucleated cells (≥3 nuclei). (E) The effect of SSD on osteoclast formation in a time-dependent manner. BMMs which were stimulated by 50ng/mL RANKL and treated with 2μM SSD from day 1 to day 3, from day 3 to day 5, or from day 5 to day 7 were fixed with 4% PFA and subjected to TRAP staining. (F) The number and area of TRAP-positive multinucleated cells (≥3 nuclei). Original magnification, ×100. Scale bar, 200 μm. Mean ± SD is used to describe the data. Above experiments were conducted independently at least 3 times. ****P <0.0001, ***P<0.001, **P <0.01.