eng Oriental Scientific Publishing Company Biomedical and Pharmacology Journal 0974-6242 2024-09-30 17 3 1749 1755 10.13005/bpj/2980 60106 article Agnis Pondineka Ria Aditama 1 Dewi Taurisiawati Rahayu 1 Brivian Florentis Yustanta 2 Wuri Widi Astuti 2 Wahyu Wijayati 2 Estin Gita Maringga 2 Health Polytechnic of Jember, Jember, Indonesia. Department of Midwifery, STIKES Karya Husada Kediri, Kediri, Indonesia Women over the age of 40 often experience a decrease in bone mass due to a lack of estrogen, which can increase the risk of osteoporosis. Previous studies have shown that the leaves of Green Clover (Marsilea crenata C. Presl.) contains phytoestrogen compounds which can potentially replace the function of estrogen in the body. In this context, this study aims to evaluate the ability of water fractions from M. crenata leaves in increasing the expression of osterix (Osx) and osteocalcin (Ocn), which are important indicators of increased bone formation, specificially on differentiation and maturation of osteoblast cell. In this study, the water fraction with a dose of 62.5; 125; and 250 µg/L, as well as Genistein 2.5 µg/ml, were given to human fetal osteoblast (hFOB) 1.19 cells that had reached the confluence point. The analysis method uses immunocytochemical techniques with confocal laser scanning microscopy (CLSM) to identify changes in the bone formation process. All dosages of water fraction considerably enhanced the bone formation in hFOB 1.19 cells, as indicated by the results. The results of this study provide confirmation that the water fraction derived from M. crenata leaves has promise as a possible agent for combating osteoporosis. This is achieved by enhancing differentiation and maturation of osteoblast cell through the upregulation of Osx and Ocn genes. https://biomedpharmajournal.org/vol17no3/the-effect-of-water-fraction-derived-from-green-clover-marsilea-crenata-c-presl-leaves-on-differentiation-and-maturation-of-human-osteoblast-cell/ Differentiation in vitro Maturation Marsilea crenata C. Presl. Osteoblast Osteocalcin Osterix