Agnis Pondineka Ria Aditama - Health Polytechnic of Jember, Jember, Indonesia
Fitria Nugrahaeni - Universitas Muhammadiyah Prof. DR. HAMKA, Jakarta, Indonesia
Dwi Susiloningrum - Departement of Pharmacy, Institut Teknologi Kesehatan Cendekia Utama Kudus, Kudus, Indonesia
Dewi Perwito Sari - Department of Pharmacy, Faculty of Health Science, PGRI Adi Buana University, Surabaya, Indonesia
Ginanjar Putri Nastiti - Department of Pharmacy, Faculty of Medicine and Health Science, Maulana Malik Ibrahim State Islamic University, Malang, Indonesia
Muhajirin Dean - Department of Pharmacy, Faculty of Medicine dan Veterinary Medicine, Nusa Cendana University, Kupang, Indonesia
Burhan Ma'arif - Department of Pharmacy, Faculty of Medicine and Health Science, Maulana Malik Ibrahim State Islamic University, Malang, Indonesia

Estrogen is important for increasing bone formation and preventing bone resorption. phytoestrogens are groups of chemicals that have structures or functions similar to estrogen. They can take the place of estrogen in the body and have a direct effect on bone density by activating estrogen-β receptors (ER-β*) and encouraging the expression of osteocalcin (Ocn). The leaves of Marsilea crenata C. Presl. are known to contain phytoestrogens. This research aimed to determine the effect of the ethyl acetate fraction of M. crenata leaves (EFM) on increasing the ER-β* by measuring free estrogen receptor-β (ER-β) expression as well as increasing osteocalcin (Ocn) expression in the human fetal osteoblastic (hFOB) ۱.۱۹ cell line. This research was carried out by giving the EFM at doses of ۶۲.۵, ۱۲۵, and ۲۵۰ ppm. Genistein ۱μM is used as a positive control. Expression of ER-β and Ocn was analyzed using the immunocytochemistry (ICC) method and a confocal laser scanning microscopy (CLSM) instrument. The results showed that EFM optimally reduced ER-β expression at ۲۵۰ ppm by ۹۰۹.۸۰ arbitrary unit (AU) and increased Ocn expression at ۱۲۵ ppm by ۷۳۴.۹۴ AU. Further research in M. crenata is necessary because it suggests that the phytoestrogen content of the EFM may play a role in bone formation and have the potential to become an antiosteoporosis alternative drug.

Bone Formation, ER-β, Herbal medicine, Ocn, phytoestrogens