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Extracellular Vesicles Carrying RUNX3 Promote Differentiation of Dental Pulp Stem Cells

  • Original Article
  • Published:
Tissue Engineering and Regenerative Medicine Aims and scope

Abstract

Background:

This study aims to clarify the mechanism underlying dental pulp cells-extracellular vesicles (DPC–EVs) carrying runt-related transcription factor 3 (RUNX3) in mediating odontogenic differentiation of dental pulp stem cells (DPSCs) with the involvement of miR-30a-5p-regulated NOTCH1.

Methods:

Extracellular vesicles (EVs) were isolated from human DPSCs, and identified using transmission electron microscopy, and nanoparticle tracking analysis. PBS, EVs, or EV inhibitor GW4869 was added to DPSCs for co-culture, whilst odontogenic differentiation was assessed in terms of ratio of mineralized nodules and expression odontoblast differentiation markers. Dual luciferase reporter gene assay and chromatin immunoprecipitation for binding relation among RUNX3, miR-30a-5p and NOTCH1were employed to evaluate their roles in odontogenic differentiation was determined. Animal experiment was established to confirm the effect of DPC-EVs-loaded RUNX3 on dental pulp.

Results:

In vitro finding demonstrated that EVs delivered RUNX3 to DPSCs, thereby activated miR-30a-5p expression and inhibited NOTCH1 expression, which was reversed by addition of GW4869. RUNX3 upregulation promoted miR-30a-5p while miR-30a-5p targeted and inhibited NOTCH1. Silencing of RUNX3 in EVs decreased expression of those differentiation markers, downregulated miR-30a-5p and upregulated NOTCH1.

Conclusion:

DPSC-EVs can carry RUNX3 to the DPSCs, promote the transcription of miR-30a-5p, and then inhibit the expression of NOTCH1, and finally promote the odontogenic differentiation of DPSCs.

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Data availability statement

The datasets generated and/or analysed during the current study are available in the manuscript and supplementary materials.

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Acknowledgement

We would like to give our sincere appreciation to the reviewers for their helpful comments on this article.

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Authors and Affiliations

  1. Department of Stomatology, The People’s Hospital of Suzhou New District, 16-502, Dongbang Xinyuan, Fengqiao Street, Huqiu District, Suzhou, Jiangsu Province, People’s Republic of China

    Yuhong Chi, Tingzhong Liu & Hao Liu

  2. Department of Oral and Maxillofacial Surgery, The First Hospital of Qiqihar, Affiliated Qiqihar Hospital of Southern Medical University, Qiqihar, People’s Republic of China

    Qingsong Jin

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  1. Yuhong Chi
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Correspondence to Hao Liu.

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Ethical statement

This study was approved by the was approved by the Ethics Committee of The People's Hospital of Suzhou New District. All procedures performed in studies were in accordance with the ethical standards of the Ethics Committee of The People's Hospital of Suzhou New District (approval number: 2022-027) and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards. All patients have ambush teeth, and provided informed consent to use their teeth for scientific research. All nude mice experiments are in accordance with the agreement approved by the UIC Animal Care Committee. This study is reported in accordance with ARRIVE guidelines (https://arriveguidelines.org).

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Chi, Y., Liu, T., Jin, Q. et al. Extracellular Vesicles Carrying RUNX3 Promote Differentiation of Dental Pulp Stem Cells. Tissue Eng Regen Med 21, 111–122 (2024). https://doi.org/10.1007/s13770-023-00578-1

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  • DOI: https://doi.org/10.1007/s13770-023-00578-1

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