Inhibition of miR-615-3p enhances dentinogenesis in scaps via PVT1-mediated mitochondrial regulation

  • Stem Cell Res Ther. 2025 Jul 31;16(1):416. doi: 10.1186/s13287-025-04528-7.
Haoqing Yang  #  1  2 ,  Yishu Huang  #  1 ,  Jiaxin Song  2 ,  Xiao Han  1 ,  Fengning Yuan  1 ,  Zhipeng Fan  3  4
Affiliations
  • 1. Laboratory of Molecular Signaling and Stem Cells Therapy, Beijing Key Laboratory of Tooth Regeneration and Function Reconstruction, Capital Medical University School of Stomatology, No. 4 Tiantanxili, Dongcheng District, Beijing, China.
  • 2. Outpatient Department of Oral and Maxillofacial Surgery, School of Stomatology, Capital Medical University, Beijing, China.
  • 3. Laboratory of Molecular Signaling and Stem Cells Therapy, Beijing Key Laboratory of Tooth Regeneration and Function Reconstruction, Capital Medical University School of Stomatology, No. 4 Tiantanxili, Dongcheng District, Beijing, China. [email protected].
  • 4. Beijing Laboratory of Oral Health, Capital Medical University, Beijing, China. [email protected].
  • # Contributed equally.
PMID: 40745571 DOI: 10.1186/s13287-025-04528-7
Abstract

Background: Mesenchymal stem cells (MSCs) are critical for dental tissue regeneration, yet their differentiation potential is tightly regulated by MicroRNAs (miRNAs). This study aimed to investigate the role of miR-615-3p in regulating odontogenic differentiation in stem cells from the apical papilla (SCAPs), offering insights into potential applications for enhancing dental tissue regeneration and repair.

Methods: Quantitative PCR (qPCR), Western blot analysis, Alkaline Phosphatase (ALP) activity assay, and Alizarin Red staining (ARS) were performed to assess odontogenic differentiation following miR-615-3p modulation in SCAPs. Mitochondrial function was evaluated by measuring Reactive Oxygen Species (ROS) levels, membrane potential, and respiratory activity. In vivo, SCAPs with miR-615-3p modulation were transplanted into rabbit extraction sockets to examine dentin-like tissue formation.

Results: miR-615-3p was significantly downregulated in SCAPs compared to umbilical cord mesenchymal stem cells (WJCMSCs) and further decreased during mineralization induction, suggesting its negative regulatory role in odontogenic differentiation. Inhibition of miR-615-3p enhanced ALP activity, mineralization, and odontogenic marker expression both in vitro and in vivo. Proteomic analysis revealed that miR-615-3p inhibition improved mitochondrial function by reducing ROS levels and increasing mitochondrial function. Further Competing Endogenous RNA Sequencing(ceRNA-seq) analysis identified PVT1 as a downstream target of miR-615-3p. PVT1 overexpression promoted odontogenic differentiation and mitochondrial homeostasis, while its knockdown impaired these processes. Collectively, the miR-615-3p/PVT1 axis emerged as a critical regulator of dentinogenesis through mitochondrial modulation.

Conclusions: Inhibiting miR-615-3p fosters dentinogenesis through PVT1-mediated mitochondrial regulation in SCAPs. These findings highlight the miR-615-3p/PVT1 axis as a promising target for enhancing dentin tissue engineering applications.

Keywords
MCSs; Mitochondrial function; Odontogenic differentiation; PVT1; miR-615-3p.
Products
  • Cat. No.
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    Description
    Target
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  • HY-D0985A
    TMRE
    98.49%, Mitochondrial Membrane Potential Fluorescent Dye
    target: Fluorescent Dye
    Research Areas: Others