MiR-376a-3p inhibits bone repair by regulating osteoblastic differentiation

  • J Orthop Surg Res. 2025 Oct 13;20(1):887. doi: 10.1186/s13018-025-06291-0.
Jian Zhang  #  1 ,  Jinxiang Zhang  #  2  3 ,  Fangyuan Zhang  4 ,  Shaochun Jing  5 ,  Debing Yang  6
Affiliations
  • 1. Department of Orthopedics, Affiliated Hospital of Yangzhou University, Yangzhou, 225009, China.
  • 2. Department of Joint Surgery and Geriatric Orthopedics, The Affiliated Hospital of Youjiang Medical University for Nationalities, Baise, 533000, China.
  • 3. Guangxi Key Laboratory for Preclinical and Translational Research on Bone and Joint Degenerative Diseases, Baise, 533000, China.
  • 4. Department of Trauma Surgery, First Hospital of Qinhuangdao, Qinhuangdao, 066000, China.
  • 5. Department of Traumatic Arthropathy, The Second Hospital of Qinhuangdao, Qinhuangdao, 066000, China.
  • 6. Department of Orthopedics, Sinopharm Southwest Aluminium Hospital, No. 15, Xihua Road, Xipeng Town, Jiulongpo District, Chongqing, 401326, China. [email protected].
  • # Contributed equally.
PMID: 41084006 DOI: 10.1186/s13018-025-06291-0
Abstract

Background: Nonunion occurs in about 10% of fracture cases, which leads to secondary surgical injury and economic burden for patients.

Purpose: Our study aimed to confirm the effects of miR-376a-3p/WWTR1 on fracture healing.

Methods: Through bioinformatics analysis, we found that miR-376a-3p was highly expressed in patients with fracture nonunion. We confirmed the interaction of miR-376a-3p and WWTR1 by using RT-qPCR, RNA immunoprecipitation, and Dual-Luciferase assay. Rescue experiment was used to explore the effects of miR-376a-3p/WWTR1 on human bone marrow mesenchymal stem cells (hBMSCs) differentiation into osteoblasts.

Results: We confirmed that miR-376a-3p was up-regulated in fracture nonunion patients by a retrospective study of clinical data of fracture patients. In vitro experiments showed that inhibiting the miR-376a-3p would increase the expression of WWTR1. It was found in the induction of the hBMSCs differentiation assay that inhibition of miR-376a-3p or overexpression of WWTR1 could increase the level of ALP, RUNX2, and OSX in cell supernatant, which reflected the increased differentiation of hBMSCs into osteoblasts. And rescue experiment confirmed that miR-376a-3p inhibits differentiation of hBMSCs through targeted regulating WWTR1.

Conclusion: We revealed the regulatory role of miR-376a-3p/WWTR1 in fracture nonunion by inhibiting osteogenic induction, which provides the possibility that miR-376a-3p may be a new underlying target for fracture nonunion prediction and therapy.

Keywords
WWTR1; Bone marrow mesenchymal stem cells; Fracture nonunion; miR-376a-3p.
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