Objectives : In traditional Korean medicine, osteoporosis is not viewed merely as a reduction in bone mass but is understood as a state of disharmony of the whole body, particularly with the decline in bone quality being closely associated with kidney deficiency. Polygonati Odorati Rhizoma (POR) has traditionally been reported to have anti-inflammatory properties, and based on its ability to nourish yin (陰, yīn) and generated body fluids (津液, jīn yè), it may be effective for bone disease related to yin deficiency. However, no studies have yet been reported on the anti-osteoporosis effects of POR. Therefore, this study investigated the effects of POR extract on the differentiation and function of osteoclast. Methods : To evaluate the inhibitory effect of Polygonati Odorati Rhizoma water extract (W_POR) and 30% ethanol extract (Et_POR) were used. RAW 264.7 cells were co-treated with receptor activator of nuclear factor-κB ligand (RANKL) and each extract, and the inhibitory effects were compared using tartrate-resistant acid phosphatase (TRAP) staining and TRAP activity assays. The mechanism was analyzed by Western blot and RT-PCR to examine changes in nuclear factor of activated T-cells 1 (NFATc1) and cellular proto-oncogene Fos (c-Fos) expression, and RT-PCR was used to evaluate the expression of genes related to osteoclast fusion, differentiation and bone resorption. Results : Et_POR significantly reduced the number and size of TRAP-positive cells compared with W_POR, demonstrating a superior inhibitory effect on osteoclast differentiation. Et_POR suppressed the expression of NFATc1 and c-Fos at the protein and gene levels, and also reduced the expression of genes related to osteoclast differentiation and bone resorption, such as TRAP, osteoclast-associated receptor (OSCAR), ATPase H+ transporting V0 subunit d2 (Atp6v0d2), carbonic anhydrases II (CA2) and matrix metallopeptidase 9 (MMP-9). Conclusions : These results indicate that POR effectively inhibits osteoclast differentiation induced by RANKL through the NFATc1/c-Fos pathway. This suggests that POR has pharmacological potential to regulate osteoclast formation and function, and may be used as a potential therapeutic agent for bone resorption disease such as osteoporosis.