Effect of Steroidal Hormone Pregnenolone on Proliferation and Differentiation of MC3T3-E1 Osteoblast like Cells

Abstract

Background: Bone remodeling is a complex process which includes continuous resorption by osteoclast cells and bone formation by osteoblast cells. Bone fragility is a common issue of elderly population particularly in postmenopausal women. It has been established that steroidal hormones have important role in bone homeostasis. Therefore hormone replacement therapy could have beneficial effects on bone health as compared to other treatments. Objectives: Imbalance between the rate of bone formation and bone resorption leads to fragility of bones. During the current study, we aimed to explore the ability of pregnenolone (1) (PRE), on proliferation and differentiation of MC3T3-E1 cells. We further aimed to investigate the underlying mechanism of action for anabolic effect of PRE (1). Methods: Effects of pregnenolone (1) on proliferation, differentiation, and mineralization of MC3T3 osteoblast like cells were determined .Cell viability was analyzed using MTT assay, and flow cytometry. ALP activity and alizarin staining were employed to evaluate the effect of pregnenolone on osteoblast differentiation. Moreover, western blot for analysis of certain important proteins, crucial for regulation of bone homeostasis, such as BMP2 and RANKL, was also performed. Results: Our results showed that pregnenolone (1) at a concentration of 5 μM caused a significant (p< 0.05) rise in the growth of MC3T3-E1 cells, whereas a comparable effect was observed in osteoblast differentiating assays. Significant decrease in RANKL expression was observed at (0.04 – 1 µM). Our results therefore indicated the possible role of pregnenolone (1) in positive regulation of bone homeostasis by suppressing RANKL expression. Conclusion: Taken together, our results indicate that pregnenolone (1) has the potential to enhance osteoblast proliferation, as inferred from the increased number of cells. These results demonstrated that pregnenolone (1) could be a potential anabolic agent for the treatment of fragility related disorders.