Department of Dental and Biomedical Materials Science, Graduate School of Biomedical Sciences, Nagasaki University, 1-7-1 Sakamoto, Nagasaki 852-8588, Japan.
Department of Applied Prosthodontics, Institute of Biomedical Sciences, Nagasaki University, 1-7-1 Sakamoto, Nagasaki 852-8588, Japan.
Int J Mol Sci. 2021 Nov 12;22(22):12259. 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.
最近,铁死亡作为一种与铁相关的调节性坏死引起了科学家们的关注。然而,关于铁死亡对骨骼影响的研究报告并不多。因此,本研究通过使用铁死亡抑制剂 ferrostatin-1 来评估铁死亡对 MC3T3-E1 前成骨细胞的影响。采用细胞图像、细胞活力、碱性磷酸酶活性试验、茜素红染色和实时 PCR 检测 RUNX2 基因表达等方法,研究 ferrostatin 和 erastin 对 MC3T3-E1 成骨细胞的影响。Erastin 被用作一种著名的铁死亡诱导剂试剂。使用浓度范围为 0 至 50µmol/L 的不同浓度 erastin 诱导细胞死亡。25µmol/L 的 erastin 导致成骨细胞发生可控性的部分细胞死亡。用 0 至 40µmol/L 的 ferrostatin-1 进行细胞掺杂和细胞死亡抑制作用。Ferrostatin-1 对已接受 erastin 部分人工细胞死亡的样本也显示出恢复效果。细胞分化、茜素红染色和 RUNX2 基因表达证实了 ferrostatin-1 对成骨细胞的骨形成能力的促进作用。本研究旨在评估 ferrostatin-1 基于其铁死亡抑制特性对 MC3T3-E1 成骨细胞系的影响。