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

• Published in: International journal of molecular sciences Vol. 22; no. 22; p. 12259
• Main Authors: Valanezhad, Alireza, Odatsu, Tetsurou, Abe, Shigeaki, Watanabe, Ikuya
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 12.11.2021; MDPI
• Subjects: Alizarin; Alkaline phosphatase; Alkaline Phosphatase - metabolism; Animals; Apoptosis; Biomedical materials; bone formation; Bone growth; Cancer; Cbfa-1 protein; Cell death; Cell differentiation; Cell growth; Cell Line; Cell Proliferation - drug effects; Cell Survival - drug effects; Cell viability; Core Binding Factor Alpha 1 Subunit - genetics; Cyclohexylamines - pharmacology; Differentiation (biology); erastin; Extracellular matrix; Ferroptosis; Ferroptosis - drug effects; ferrostatin-1; Gene expression; Gene Expression - drug effects; Homeostasis; Iron; Lipid peroxidation; Lipids; Metabolism; Mice; Mineralization; Mitochondria; Necrosis; Osteoblasts; Osteoblasts - cytology; Osteoblasts - drug effects; Osteoblasts - metabolism; Osteogenesis; Osteogenesis - drug effects; Osteoporosis; Phenylenediamines - pharmacology; Piperazines - pharmacology; Reagents; regulated necrosis; Reverse Transcriptase Polymerase Chain Reaction; Staining; Transcription factors; erastin; bone formation; ferrostatin-1; ferroptosis; regulated necrosis
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms222212259

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.