Nanoemulsified Genistein and Vitamin D Mediated Epigenetic Regulation to Inhibit Osteoporosis

Abstract

The imbalance in the bone remodeling process with more bone resorption by osteoclasts compared to bone formation by osteoblasts results in a metabolic bone disorder known as osteoporosis. This condition reduces the bone mineral density and increases the risk of fractures due to low bone mass and disrupted bone microarchitecture. Osteoclastogenesis increases when the receptor activator NFκB ligand (RANKL) on the osteoblast surface binds to the receptor activator NFκB (RANK) on the osteoclast surface and the function of the decoy receptor of RANKL, osteoprotegrin, is compromised due to external stimuli such as heparin and lipopolysaccharides. The RANK/RANKL axis promotes the nuclear factor kappa B (NFκB) expression, which in turn increases the histone methyltransferase activity of EzH2 and EzH1 for the epigenetic regulation of osteoclastogenesis-related genes. Genistein counteracts NFκB-induced osteoclastogenesis and downstream signaling through the direct regulation of histone methyltransferase, EzH2 and EzH1, transcription. However, genistein possesses limitations like low bioavailability, low water solubility, high estrogen activity, and thyroid side effects, which obstruct its therapeutic usage. Here, the nanoemulsified formulation of genistein with vitamin D was utilized to circumvent the limitations of genistein so that it can be utilized for therapeutic purposes in osteoporosis management. The nanoemulsification of genistein and vitamin D was performed through the spontaneous emulsification using Tween 80 and medium chain triglyceride oil as an organic phase. The physiologically stable and biocompatible combination of the genistein and vitamin D nanoemulsion (GVNE) exhibited the controlled release pattern of genistein with Korsmeyer−Peppas and Higuchi models under different pH conditions (7.4, 6.5, and 1.2). The GVNE potentially enhanced the therapeutic efficacy under in vitro osteoporosis models and helped restore disease parameters like alkaline phosphatase activity, tartrate-resistant acid phosphatase activity, and the formation of multinuclear giant cells. Molecularly, the GVNE overturned the LPS-induced osteoclastogenesis by downregulation of NFκB expression along with its binding on EzH2 and EzH1 promoters. GVNE effects on the osteoporosis model established it as an efficient antiosteoporotic therapy. This nanonutraceutical-based formulation provides an epigenetic regulation of osteoporosis management and opens new avenues for alternate epigenetic therapies for osteoporosis.

Targeting TFF3 in obstructive airway diseases: a computational approach to novel therapeutics

“…Derived from soybeans, this iso avone has been extensively researched for its myriad health bene ts, ranging from its antioxidant and anti-in ammatory properties to its demonstrated antiproliferative effects on diverse cancer cell lines [26][27][28][29]. Additionally, Genistein has been shown to have a potential role in the prevention and treatment of osteoporosis by increasing bone mineral density [30]. Genistein has also been shown to have potential bene ts in the treatment of cardiovascular disease, diabetes, and neurodegenerative disorders [31][32][33].…”

Noninvasive and Microinvasive Nanoscale Drug Delivery Platforms for Hard Tissue Engineering

“…Thus, its deficiency leads to osteomalacia in adults and rickets in children; it also contributes to osteopenia and osteoporosis. Research studies for enhancing the bioavailability of cholecalciferol and protecting it from the acidic pH of the GIT have been performed using nanoencapsulation by ultrasonication or via standard emulsification approach as a co-nanoemulsion . The study involving ultrasonication resulted in a high encapsulation efficiency of over 95%, but in vivo assays have not been performed.…”

“…Research studies for enhancing the bioavailability of cholecalciferol and protecting it from the acidic pH of the GIT have been performed using nanoencapsulation by ultrasonication 104 or via standard emulsification approach as a co-nanoemulsion. 105 The study involving ultrasonication resulted in a high encapsulation efficiency of over 95%, but in vivo assays have not been performed. The second study involved a formulation containing medium chain triglycerides (MCT) oil as a medium and a combination of cholecalciferol and genistein (a polyphenolic isoflavone, that along with its many other effects, 106,107 also stimulates bone formation and inhibits resorption, thus positively affecting osteoporosis and osteoarthritis 108 ).…”

A mechanistic review of chinese medicine polyphenols on bone formation and resorption

“…At the level of epigenetic modification, genistein counteracts NF-κB-induced osteoclast generation and downstream signaling by directly regulating the transcription of histone methyltransferases EzH2 and EzH1 ( Kushwaha et al, 2022 ). Furthermore, there are clinical trials showing that supplementation with the dietary phytoestrogen genistein may be as effective as hormone replacement therapy in reducing bone loss associated with menopause without the associated bone loss side effects ( Cotter and Cashman, 2003 ; Sansai et al, 2020 ).…”

“…Furthermore, there are clinical trials showing that supplementation with the dietary phytoestrogen genistein may be as effective as hormone replacement therapy in reducing bone loss associated with menopause without the associated bone loss side effects ( Cotter and Cashman, 2003 ; Sansai et al, 2020 ). In this study, to improve the bioavailability of genistein and reduce its side effects, the nanofied formulation of genistein with Vitamin D was invented to enhance the therapeutic efficacy of the osteoporosis model in vitro and improve alkaline phosphatase activity and multinucleated giant cell formation ( Kushwaha et al, 2022 ). If the bioavailability of genistein is improved, its future market development potential is huge.…”