Tehran University of Medical Sciences Acta Medica Iranica 0044-6025 56 4 2018 04 09 234 243 EN Hnin Ei Thu Department of Pharmacology, Faculty of Medicine, Universiti Kebangsaan Malaysia, Jalan Yaacob Latif 56000, Cheras, Malaysia. Zahid Hussain Department of Pharmaceutics, Faculty of Pharmacy, Universiti Teknologi MARA (UiTM), Puncak Alam Campus, Bandar Puncak Alam 42300, Selangor, Malaysia. Isa Naina Mohamed Department of Pharmacology, Faculty of Medicine, Universiti Kebangsaan Malaysia, Jalan Yaacob Latif 56000, Cheras, Malaysia. Ahmad Nazrun Shuid Department of Pharmacology, Faculty of Medicine, Universiti Kebangsaan Malaysia, Jalan Yaacob Latif 56000, Cheras, Malaysia. 2017 05 17 2017 08 13 Numerous in vitro, in vivo and clinical studies have evidenced the outstanding potential of dihydrotestosterone (DHT) in the treatment of male osteoporosis. Despite of promising clinical efficacy of DHT in regulating the skeletal growth and homeostasis, the exact molecular and translational mechanism is yet to be explored. This study was aimed to investigate the bone-forming molecular mechanism of DHT using MC3T3-E1 cell line as in vitro model. The mechanism of bone-forming ability of DHT was assessed by evaluating the time-mannered expression of bone-related biomarkers such as bone morphogenic protein-2 (BMP-2), alkaline phosphatase (ALP), runt-related transcription factor 2 (Runx-2), osteocalcin (OCN), type I collagen, osteopontin (OPN), transforming growth factor-β1 (TGF-β1) and androgen receptor (AR). Results demonstrated a remarkable efficacy of DHT (at a dose of 0.1 ng/mL) in promoting the expression of these vital bone-forming mediators. The resulting analysis revealed that the DHT-0.1 group showed higher expression of BMP-2 (106±9 pg/mL), ALP (381±16 pg/mL), Runx-2 (664±32 pg/mL), OCN (2265±111 pg/mL), type I collagen (276±16 pg/mL), TGF-β1 (81±7 pg/mL) and AR (411±21 pg/mL) compared to the control (CN) and other DHT-treated groups. These findings provide an in vitro evidence for the bone-forming capacity of DHT and its therapeutic significance for the treatment of male osteoporosis. https://acta.tums.ac.ir/index.php/acta/article/view/6468 https://acta.tums.ac.ir/index.php/acta/article/download/6468/5074