2. Academic Validation
  3. Magnesium ions facilitate osteogenic differentiation and intervertebral fusion via m6A methylation of RhoA mRNA

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

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.

Figures
Products