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

  • J Orthop Translat. 2026 Feb 18:57:101056. doi: 10.1016/j.jot.2026.101056.
Haocheng Xu  1 ,  Linli Li  1 ,  Fan Zhang  1 ,  Minghao Shao  1 ,  Chenyan Li  1 ,  Yitong Xue  1 ,  Dachuan Li  1 ,  Zhidi Lin  1 ,  Zhaoyang Gong  1 ,  Jiongdong Wu  1 ,  Zhiwen Luo  2 ,  Zhicai Shi  3 ,  Xinlei Xia  1 ,  Hongli Wang  1 ,  Xiaosheng Ma  1 ,  Jianyuan Jiang  1 ,  Xiaochuan Gu  3 ,  Yang Liu  4 ,  Xiao Lu  1 ,  Feizhou Lyu  1
Affiliations
  • 1. Department of Orthopedics, Huashan Hospital, No. 12, Middle Wulumuqi Road, Jing'an District, Fudan University, Shanghai, 200040, China.
  • 2. Department of Sports Medicine, Huashan Hospital, No. 12, Middle Wulumuqi Road, Jing'an District, Fudan University, Shanghai, 200040, China.
  • 3. Department of Orthopedics, Shanghai Changhai Hospital, Navy Medical University, Shanghai, 200433, China.
  • 4. Department of Orthopedics, Shanghai Changzheng Hospital, Naval Medical University, Shanghai, 200003, China.
PMID: 41757292 DOI: 10.1016/j.jot.2026.101056
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.

Keywords
Intervertebral fusion; METTL3; Magnesium ions; Osteogenic differentiation; RhoA/ROCK pathway; m6A modification.
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