Acta Medica Iranica 2018. 56(4):234-243.

Mechanistic Insight Into the Efficient Osteogenic Potential of Dihydrotestosterone: Exploring Sequential Expression of Bone-Related Protein Biomarkers
Hnin Ei Thu, Zahid Hussain, Isa Naina Mohamed, Ahmad Nazrun Shuid


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.


Dihydrotestosterone; Osteoporosis; Enhanced osteogenesis; Cell proliferation; Bone-related biomarkers; Time mannered expression

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Beloti MM, Rosa AL. Osteoblast differentiation of human bone marrow cells under continuous and discontinuous treatment with dexamethasone. Braz Dent J 2005;16:156-61.

Karsenty G, Wagner EF. Reaching a genetic and molecular understanding of skeletal development. Dev Cell 2002;2:389-406.

Lian JB, Javed A, Zaidi SK, Lengner C, Montecino M, van Wijnen AJ, et al. Regulatory controls for osteoblast growth and differentiation: the role of Runx/Cbfa/AML factors. Crit Rev Eukaryot Gene Expr 2004;14:1-41.

Leder BZ, LeBlanc KM, Schoenfeld DA, Eastell R, Finkelstein JS. Differential effects of androgens and estrogens on bone turnover in normal men. J Clin Endocrinol Metab 2003;88:204-10.

Van Pottelbergh I, Goemaere S, Zmierczak H, Kaufman JM. Perturbed sex steroid status in men with idiopathic osteoporosis and their sons. J Clin Endocrinol Metab 2004;89:4949-53.

Lips P. Vitamin D physiology. Prog Biophys Mol Biol 2006;92:4-8.

Clarke BL, Khosla S. New selective estrogen and androgen receptor modulators. Curr Opin Rheumatol 2009;21:374-9.

Zallone A. Direct and indirect estrogen actions on osteoblasts and osteoclasts. Ann N Y Acad Sci 2006;1068:173-9.

Maclaughlin EJ, Sleeper RB, McNatty D, Raehl CL. Management of age-related osteoporosis and prevention of associated fractures. Ther Clin Risk Manag 2006;2:281-95.

Saadiah Abdul Razak H, Shuid AN, Naina Mohamed I. Combined Effects of Eurycoma longifolia and Testosterone on AndrogenDeficient Osteoporosis in a Male Rat Model. Evid Based Complement Alternat Med 2012;2012:872406.

Bland R. Steroid hormone receptor expression and action in bone. Clin Sci (Lond) 2000;98:217-40.

Compston JE. Sex steroids and bone. Physiol Rev 2001;81:419-47.

Huang CK, Lai KP, Luo J, Tsai MY, Kang HY, Chen Y, et al. Loss of androgen receptor promotes adipogenesis but suppresses osteogenesis in bone marrow stromal cells. Stem Cell Res 2013;11:938-50.

Antonio C, Alberto F. Male osteoporosis and androgenic therapy: from testosterone to SARMs. Clin Cases Miner Bone Metab 2009;6:229-33.

Cilotti A, Falchetti A. Male osteoporosis and androgenic therapy: from testosterone to SARMs. Clin Cases Miner Bone Metab 2009;6:229-33.

Wilt TJ, MacDonald R, Hagerty K, Schellhammer P, Kramer BS. Five-alpha-reductase Inhibitors for prostate cancer prevention. Cochrane Database Syst Rev 2008;2:CD007091.

Kapur SP, Reddi AH. Influence of testosterone and dihydrotestosterone on bone-matrix induced endochondral bone formation. Calcif Tissue Int 1989;44:108-13.

Thu HE, Mohamed IN, Hussain Z, Shuid AN. Dihydrotestosterone, a robust promoter of osteoblastic proliferation and differentiation: understanding of time-mannered and dose-dependent control of bone forming cells. Iran J Basic Med Sci 2017;20:894-904.

Lin IC, Slemp AE, Hwang C, Sena-Esteves M, Nah HD, Kirschner RE. Dihydrotestosterone stimulates proliferation and differentiation of fetal calvarial osteoblasts and dural cells and induces cranial suture fusion. Plast Reconstr Surg 2007;120:1137-47.

Bart L. Clarke, Sundeep Khosla. Androgens and Bone. Steroids 2009;74:296-S305.

Sinnesael M, Boonen S, Claessens F, Gielen E, Vanderschueren D. Testosterone and the male skeleton: a dual mode of action. J Osteoporos 2011;2011:240328.

Yarrow JF, Wronski TJ, Borst SE. Testosterone and Adult Male Bone: Actions Independent of 5α-Reductase and Aromatase. Exerc Sport Sci Rev 2015;43:222-30.

Balkan W, Burnstein KL, Schiller PC, Perez-Stable C, D'Ippolito G, Howard GA, et al. Androgen-induced mineralization by MC3T3-E1 osteoblastic cells reveals a critical window of hormone responsiveness. Biochem Biophys Res Commun 2005;328:783-89.

Luppen CA, Leclerc N, Noh T, Barski A, Khokhar A, Boskey AL, et al. Brief bone morphogenetic protein 2 treatment of glucocorticoid-inhibited MC3T3-E1 osteoblasts rescues commitment-associated cell cycle and mineralization without alteration of Runx2. J Biol Chem 2003;278,44995-5003.

Yamaguchi A, Komori T, Suda T. Regulation of osteoblast differentiation mediated by bone morphogenetic proteins, hedgehogs, and Cbfa1. Endocr Rev 2000;21:393-411.

Raida M, Heymann AC, Gunther C, Niederwieser D. Role of bone morphogenetic protein 2 in the crosstalk between endothelial progenitor cells and mesenchymal stem cells. Int J Mol Med 2006;18:735-9.

Ripamonti U, Crooks J, Matsaba T, Tasker J. Induction of endochondral bone formation by recombinant human transforming factor‐2 in the baboon. Growth Factors 2000;17:269-85.

Suzawa M, Takeuchi Y, Fukumoto S, Kato S, Ueno N, Miyazono K, et al. Extracellular matrix associated bone morphogenetic proteins are essential for differentiation of murine osteoblastic cells in vitro. Endocrinology 1999;140:2125-33.

Nakano Y, Addison WN, Kaartinen MT. ATP induced mineralization of MC3T3-E1 osteoblast cultures. Bone 2007;41:549-61.

Jeong JC, Lee JW, Yoon CH, Kim HM, Kim CH. Drynariae Rhizoma promotes osteoblast differentiation

and mineralization in MC3T3-E1 cells through regulation of bone morphogenetic protein-2, alkaline phosphatase, type I collagen and collagenase-1. Toxicol in Vitro 2004;18:829-34.

Komori T. Regulation of bone development and extracellular matrix protein genes by RUNX2. Cell Tissue Res 2010;339:189-95.

Sakamoto W, Isomura H, Fujie K, Deyama Y, Kato A, Nishihira J, et al. Homocysteine attenuates the expression of osteocalcin but enhances osteopontin in MC3T3-E1 preosteoblastic cells. Biochim Biophys Acta 2005;1740:12-6.

Thu HE, Mohamed IN, Hussain Z, Shuid AN. Eurycoma longifolia as a potential alternative to testosterone for the treatment of osteoporosis: Exploring time-mannered proliferative, differentiative and morphogenic modulation in osteoblasts. J Ethnopharmacol 2017;195:143-58.

Kim HK, Kim MG, Leem KH. Osteogenic activity of collagen peptide via ERK/MAPK pathway mediated boosting of collagen synthesis and its therapeutic efficacy in osteoporotic bone by back-scattered electron imaging and microarchitecture analysis. Molecules 2013;18:15474-89.

Huang W, Carlsen B, Rudkin G, Berry M, Ishida K, Yamaguchi DT, et al. Osteopontin is a negative regulator of proliferation and differentiation in MC3T3-E1 pre-osteoblastic cells. Bone 2004;34:799-808.

Ihara H, Denhardt DT, Furuya K, Yamashita T, Muguruma Y, Tsuji K. et al. Parathyroid hormone-induced bone resorption does not occur in the absence of osteopontin. J Biol Chem 2001;276:13065-71.

Beck GR Jr, Zerler B, Moran E. Phosphate is a specific signal for induction of osteopontin gene expression. Proc Natl Acad Sci U S A 2000;97:8352-57.

Derynck R, Akhurst RJ. Differentiation plasticity regulated by TGFbeta family proteins in development and disease. Nat Cell Biol 2007;9:1000-4.

Guo X, Wang XF. Signaling cross-talk between TGF-beta/BMP and other pathways. Cell Res 2009;19:71-88.

Riggs BL, Khosla S, Melton LJ 3rd. Sex steroids and the construction and conservation of the adult skeleton. Endocrine Rev 2002;23:279-302.


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