Dihydrotestosterone, a robust promoter of osteoblastic proliferation and differentiation: understanding of time-mannered and dose-dependent control of bone forming cells

Objective(s): The present study was aimed to evaluate the time-mannered and dose-dependent effects of ۵α-dihydrotestosterone (۵α-DHT) on the proliferation and differentiation of bone forming cells using MC۳T۳-E۱ cells. Materials and Methods: Cell proliferation was analyzed using MTS and phase contrast microscopic assays. Osteogenic differentiation was assessed through a series of in vitro experiments including crystal violet staining, alkaline phosphatase (ALP) activity, and Van Gieson (VG) staining. Taken together, the efficiency of bone mineralization was examined by using alizarin red s (ARS) staining, Von Kossa staining, scanning electron microscopy (SEM) and energy dispersive x-ray (EDX) analysis. Results: The resulting data revealed that ۵α-DHT exhibits promising potential particularly at a dose of ۰.۱ ng/ml, in promoting the growth of MC۳T۳-E۱ cells compared to the control group (CN). Moreover, a significantly higher ALP activity was evident in the experimental group treated with ۵α-DHT compared to the CN group at various time intervals. MC۳T۳-E۱ cells treated with ۵α-DHT also expressed a remarkably higher collagen deposition and mineralization (calcium and phosphate contents) compared to the CN group at various time intervals. Conclusion: Conclusively, we suggest that ۵α-DHT exhibits outstanding potential of promoting proliferation and differentiation in osteoblasts which could be the in vitro basis for the efficacy of ۵α-DHT in the treatment of androgen-deficient male osteoporosis.

Keywords:

Active bone formation , Differentiation , MC۳T۳-E۱ cells , Morphogenic modulation , Proliferation , ۵α-dihydrotestosterone