###### Abstract  
Owing to global population aging, instances of bone metabolic diseases have increased. Consequently, interest in natural and functional plant food products for the prevention and treatment of osteoporosis is also increasing. In this study, we determine the potential therapeutic effects of _Benicasa hispida_ extract (HR1901-W) on osteoblast and osteoclast differentiation. The potential preventive effects of _Benicasa hispida_ in osteoporosis have not previously been reported. We identified and analyzed 2-furoic acid, a chemical component of HR1901-W. We evaluated whether HR1901-W promoted osteogenesis in the MC3T3-E1 cell line and whether it inhibited the differentiation of RAW 264.7 macrophage cells (osteoblast precursors). We observed that HR1901-W promoted significantly high dose-dependent proliferation and extracellular matrix mineralization in MC3T3-E1 cells. In fact, increased cell proliferation was found to be associated with increased protein expression of factors related to osteoblast differentiation, including alkaline phosphatase, osteoclast, and runt-related transcription factor 2. On the other hand, macrophage colony-stimulating factor (10 ng/mL) and nuclear factor-xB ligand (100 ng/mL) treated differentiated RAW264.7 macrophages exhibited a significant reduction in tartrate-resistant acid phosphatase activity. Taken together, our results indicate that HR1901-W is a promising candidate of functional materials that regulate the balance between bone-forming osteoblasts and bone-resorbing osteoclasts to prevent osteoporosis. Benicasa hispida]Benincasa hispida  
other minerals are deposited onto the bone or its extracellular matrix [11; 12]. Notably, the differentiation of osteoblasts is mediated by the differential expression of specific factors. For instance, runt-related transcription factor 2 (Runx2) regulates the early stages of osteoblast differentiation by promoting the differentiation of pluripotent mesenchymal cells into immature osteoblasts [13; 14]. Another key factor of osteoblast differentiation is alkaline phosphatase (ALP) that is synthesized by osteoblast precursors immediately following growth arrest [15; 16]. Notably, ALP is an ectoenzyme that is attached to the outer surface of the osteoblast cell membrane. It facilitates calcium deposition on the bone by transporting inorganic phosphoric acid during mineralization [17; 18]. While it initially forms the bone mass, it subsequently plays a role in bone remodeling and mineral metabolism [19; 20]. Osteoclasts are multinucleated cells derived from the monocyte/macrophage lineage that resorb the mineralized matrices formed by osteoblasts. The macrophage colony-stimulating factor (M-CSF) and the receptor activator of nuclear factor-xB ligand (RANKL) stimulate the differentiation of immature osteoclasts into mature osteoclasts. These factors are recruited by hematopoietic stem cells to specific regions of the bone surface. Furthermore, they initiate bone resorption by stimulating osteoclastogenesis factors in the early stage of osteoclasts [21; 22]. Tartrate-resistant acid phosphatase (TRAP) is specifically expressed when immature osteoclasts differentiate into multinucleated osteoclasts. Notably, its expression is used to confirm osteoclast proliferation [23]. _Benincasa hispida_ (Thunb.) Cogn. (syn. _Benincasa cerifera_ [PERSON]) is an annual plant of the family Cucurbitaceae. It is a vegetable widely distributed in Asian countries and regions such as Southeast Asia, India, China, and Korea [24; 25]. _B. hispida_ fruit contains 93-96% moisture and is rich in nutrients such as vitamin C, vitamin B2, Na, and Ca [17; 26; 27]. Phenolic compounds such as asstilbin, catechin, and naringenin have been isolated from _B. hispida_ fruit [28]. Other bioactive compound constituents include triterpenes (alrusenol, multififorenol), isomultiforenol), sterols (lupeol, lupeol acetate, \(\beta\)-sitosterol), glycosides, saccharides, caroten, \(\beta\)-sitosterin, tannins, and uronic acid [29; 30].