Magnesium ions facilitate osteogenic differentiation and intervertebral fusion via m6A methylation of RhoA mRNA

IF 5.9 1区医学 Q1 ORTHOPEDICS

Journal of Orthopaedic Translation Pub Date : 2026-03-01 Epub Date: 2026-02-18 DOI:10.1016/j.jot.2026.101056

Haocheng Xu , Linli Li , Fan Zhang , Minghao Shao , Chenyan Li , Yitong Xue , Dachuan Li , Zhidi Lin , Zhaoyang Gong , Jiongdong Wu , Zhiwen Luo , Zhicai Shi , Xinlei Xia , Hongli Wang , Xiaosheng Ma , Jianyuan Jiang , Xiaochuan Gu , Yang Liu , Xiao Lu , Feizhou Lyu

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引用次数: 0

Abstract

Background

Magnesium-based implants facilitate bone regeneration via degradation. However, the epigenetic mechanisms, particularly N6-methyladenosine (m6A) modification regulated by Mg²⁺, remain incompletely understood. This study investigated the role of Mg²⁺ in osteogenic differentiation through the METTL3-RhoA axis and evaluated its potential in intervertebral fusion.

Methods

The optimal Mg²⁺ concentration was identified using MC3T3-E1 cells. Methylated RNA immunoprecipitation sequencing (MeRIP-seq) and MeRIP-PCR were employed to identify m6A target genes. Functional assays (knockdown, overexpression, and rescue) validated the METTL3-YTHDF1-RhoA pathway. A rat tail intervertebral fusion model with magnesium implants was used to assess in vivo effects.

Results

Treatment with 4 mM Mg²⁺ significantly enhanced osteogenic activity and increased METTL3 levels. Mechanistically, METTL3 promoted m6A methylation of RhoA mRNA, which was subsequently bound by YTHDF1, enhancing translation and activating the RhoA/ROCK pathway. In vivo, magnesium implants accelerated fusion and improved trabecular bone quality; however, these effects were inhibited by METTL3 or RhoA inhibitors.

Conclusion

Mg²⁺ enhances osteogenic differentiation through the METTL3-YTHDF1-RhoA/ROCK pathway.

The translational potential of this article

This study provides an epigenetic framework for optimizing magnesium-based orthopedic implants and suggests that targeting the m6A-RhoA axis could improve spinal fusion outcomes.

Abstract Image

查看原文本刊更多论文

镁离子通过RhoA mRNA的m6A甲基化促进成骨分化和椎间融合。

背景：镁基种植体通过降解促进骨再生。然而，表观遗传机制，特别是Mg2+调控的n6 -甲基腺苷（m6A）修饰，仍然不完全清楚。本研究通过METTL3-RhoA轴研究了Mg2+在成骨分化中的作用，并评估了其在椎间融合中的潜力。方法：采用MC3T3-E1细胞筛选最佳Mg2+浓度。采用甲基化RNA免疫沉淀测序（MeRIP-seq）和MeRIP-PCR鉴定m6A靶基因。功能分析（敲除、过表达和挽救）验证了METTL3-YTHDF1-RhoA通路。采用大鼠尾巴椎间融合术模型对镁植入物进行体内效果评估。结果：4 mM Mg2+治疗显著增强成骨活性，增加METTL3水平。在机制上，METTL3促进RhoA mRNA的m6A甲基化，随后与YTHDF1结合，增强翻译并激活RhoA/ROCK途径。在体内，镁植入物加速融合，改善骨小梁质量；然而，这些作用被METTL3或RhoA抑制剂抑制。结论：Mg2+通过METTL3-YTHDF1-RhoA/ROCK途径促进成骨分化。本文的转化潜力：该研究为优化镁基骨科植入物提供了表观遗传学框架，并表明靶向m6A-RhoA轴可以改善脊柱融合结果。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Journal of Orthopaedic Translation Medicine-Orthopedics and Sports Medicine

CiteScore

11.80

自引率

13.60%

发文量

审稿时长

29 days

期刊介绍： The Journal of Orthopaedic Translation (JOT) is the official peer-reviewed, open access journal of the Chinese Speaking Orthopaedic Society (CSOS) and the International Chinese Musculoskeletal Research Society (ICMRS). It is published quarterly, in January, April, July and October, by Elsevier.