Genistein contributes to cell cycle progression and regulates oxidative stress in primary culture of osteoblasts along with osteoclasts attenuation

Sahabjada Siddiqui,Abbas Ali Mahdi,Md Arshad

doi:10.1186/s12906-020-03065-5

Abstract

BackgroundThe present study was designed to examine the role of isoflavone genistein (GS) on bone formation, regulating oxidative stress and cell cycle in primary osteoblasts, as well as attenuation of osteoclast formation.MethodsPrimary calvaria osteoblasts were isolated from 2 to 3 days old neonatal rat pups (n = 6–8) of Sprague Dawley rats. Osteoblasts were incubated with varying concentrations of GS and different assays viz. cell proliferation, differentiation, calcium deposition, cell cycle progression, antioxidant ability, and osteogenic gene expression were performed. Tartrate-resistant acid phosphatase (TRAP) staining and immunolocalization of cathepsin K protein were assessed in bone marrow-derived osteoclasts.ResultsResults revealed that GS markedly induced cell growth and osteoblast differentiation depending upon dose. The fluorescent dye DCFH-DA staining data proved the antioxidant ability of GS, which reduced the H2O2- induced intracellular oxidative stress in osteoblasts. Quantitative real-time PCR analysis revealed that GS treatment upregulated the expression of osteoblastic genes of Runt-related transcription factor 2 (Runx2), bone morphogenetic proteins 2 (BMP2), and osteocalcin. Immunolocalization of BMP2 also indicated the osteogenic efficacy of GS. Furthermore, TRAP staining and cathepsin K expression depicted that GS inhibited multinucleated osteoclasts formation.ConclusionsIn conclusion, GS isoflavone might impart protective effects against oxidative stress-induced bone loss and thus, could maintain skeletal growth.

Highlights

The present study was designed to examine the role of isoflavone genistein (GS) on bone formation, regulating oxidative stress and cell cycle in primary osteoblasts, as well as attenuation of osteoclast formation
Reagents and chemicals Alpha- minimum essential medium (a-MEM), foetal bovine serum (FBS), MTT powder, p-nitrophenyl phosphate, naphthol AS-MX phosphate, fast blue BB salt, ascorbic acid and Alizarin Red S dye were purchased from Himedia, India
Soy isoflavone genistein was used to its effect(s) on osteoblast proliferation and differentiation, oxidative stress and cell cycle progression in primary osteoblasts, as well as osteoclast attenuation

Summary

Introduction

The present study was designed to examine the role of isoflavone genistein (GS) on bone formation, regulating oxidative stress and cell cycle in primary osteoblasts, as well as attenuation of osteoclast formation. The balance between bone formation, bone resorption, proliferation, and apoptosis helps to maintain bone mass nearly constant [1]. An imbalance between bone resorption and bone formation may cause metabolic disorders. Siddiqui et al BMC Complementary Medicine and Therapies (2020) 20:277 foods and nutraceuticals, can be valuable assets for the peak bone mass of an individual and can support bone health during later life stages. Previous studies have shown that isoflavones and flavonoids increase bone mass density and prevent bone loss in-vivo and in-vitro [6,7,8]. Flavonoids and isoflavones have the potential to maintain and improve the bone mass of human subjects. GS is present in various food items primarily in soybeans and soy products, which is capable of interacting with various hormones like estrogens [10, 11]

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