Bone Formation Ability and Cell Viability Enhancement of MC3T3-E1 Cells by Ferrostatin-1 a Ferroptosis Inhibitor of Cancer Cells.

Alireza Valanezhad,Shigeaki Abe,Ikuya Watanabe,Tetsurou Odatsu

doi:10.3390/ijms222212259

Alireza Valanezhad, Shigeaki Abe + Show 2 more

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https://doi.org/10.3390/ijms222212259

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Abstract

Recently, ferroptosis has gained scientists’ attention as an iron-related regulated necrosis. However, not many reports have investigated the effect of ferroptosis on bone. Therefore, with the present study, we assessed the effect of ferroptosis inhibition using ferrostatin-1 on the MC3T3-E1 pre-osteoblast cell. Cell images, cell viability, alkaline phosphatase activity test, alizarin red staining, and RUNX2 gene expression using real-time PCR were applied to investigate the effects of ferrostatin and erastin on MC3T3-E1 osteoblast cells. Erastin was used as a well-known ferroptosis inducer reagent. Erastin with different concentrations ranging from 0 to 50 µmol/L was used for inducing cell death. The 25 µmol/L erastin led to controllable partial cell death on osteoblast cells. Ferrostatin-1 with 0 to 40 µmol/L was used for cell doping and cell death inhibition effect. Ferrostatin-1 also displayed a recovery effect on the samples, which had already received the partially artificial cell death by erastin. Cell differentiation, alizarin red staining, and RUNX2 gene expression confirmed the promotion of the bone formation ability effect of ferrostatin-1 on osteoblast cells. The objective of this study was to assess ferrostatin-1’s effect on the MC3T3-E1 osteoblast cell line based on its ferroptosis inhibitory property.

Highlights

Bone problems such as hip fracture and dental implant loosening triggered by osteoporosis grow rapidly due to aging worldwide
According to Dixon, S. et al (2013), in this type of cell death, lipid peroxidation plays a key role in cell membrane damage; instead of apoptosis and necrosis, the triggering of caspase and adenosine triphosphate was not noticed in ferroptosis [9]
MC3T3-E1 cell death induced by ferroptosis depends on the erastin concentration per cell number ratio

Summary

Introduction

Bone problems such as hip fracture and dental implant loosening triggered by osteoporosis grow rapidly due to aging worldwide. Understanding the mechanism of cell death in biology is essential to prevent it [1]. The idea of exclusive cell death pathways has evolved recently from apoptosis or necrosis to several other types of regulated necrosis [2]. Apoptosis is a general mechanism of controlled cell deletion, regulating cell population [3,4]. A number of regulated necrosis cases with individual pathways and functions have been introduced [8]. Ferroptosis is new regulated necrosis known as iron-dependent cell death. According to Dixon, S. et al (2013), in this type of cell death, lipid peroxidation plays a key role in cell membrane damage; instead of apoptosis and necrosis, the triggering of caspase and adenosine triphosphate was not noticed in ferroptosis [9]. Excess iron through the Fenton reaction causes ferroptosis, making hydroxyl radicals from superoxide or hydrogen peroxide [10]

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