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Berberine Promotes Osteogenic Differentiation of Human Dental Pulp Stem Cells Through Activating EGFR-MAPK-Runx2 Pathways

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Pathology & Oncology Research

Abstract

Similar to the mesenchymal stem cells (MSCs), dental pulp stem cells (DPSCs) also have pluripotent differentiation characteristic and may be more ideal for tissue regeneration, especially in tooth regeneration engineering. However, bacterial infection may be a powerful obstacle. Berberine (BBR), known with antibacterial effects, was recently found to play functions in bone formation through promoting osteogenic differentiation from pluripotent stem cells. However, whether BBR also function in DPSCs osteogenic differentiation has not yet been reported. Primary DPSCs were isolated from dental pulp tissues extracted from human impacted mandibular third molars, and identified by flow cytometry for cell surface antigen molecules. A dexamethasone osteogenic medium was used to induce DPSCs osteogenic differentiation. BBR (1 μM and 5 μM) was pre-added to into medium, and then cell proliferation, spheroid formation and osteogenic differentiation capacities of DPSCs were analyzed, as well as the underlying molecules modulation mechanism. Flow cytometry identified that CD44, CD90, CD81 and CD105 positively expressed in the isolated hDPSCs, with CD34 and CD45 negetively expressed. BBR enhanced the cell proliferation of hDPSCs in a dose-dependent pattern, and promoted dexamethasone-induced osteogenic differentiation via enhancing Runx2 transcription factor activity followed by upregulating osteogenesis markers expression, whereas the adipogenic differentiation of hDPSCs was suppressed dramatically by BBR. The EGFR and MAPK pathways were activated by BBR, and inhibitors for these pathways significantly suppressed the osteogenic differentiation promotion of BBR. These results have revealed a novel mechanism that berberine might promote hDPSCs osteogenic differentiation through activating EGFR-MAPK-Runx2 signaling pathways.

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Acknowledgments

We would like to give our sincere gratitude to the reviewers for their constructive comments.

Funding

This work was supported by Qingdao Medical Science and Technology Guiding Project (No.2017-WJZD125), Qingdao Shinan District Science and Technology Development Fund (No.2016-3-042-YY) and Qingdao Stomatological Hospital Youth Fund Project (No.2018QN01, No.2019QNJJ02).

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Authors and Affiliations

  1. Department of Cariology and Endodontology, Qingdao Stomatological Hospital, Qingdao, 266001, People’s Republic of China

    Bing-Chang Xin & De-Gang Sun

  2. Department of Stomatology, Tai’an City Central Hospital, Tai’an, 271000, People’s Republic of China

    Qi-Shan Wu & Song Jin

  3. Department of Stomatology, Guizhou Provincial People’s Hospital, Guiyang, 550002, People’s Republic of China

    Ai-Hua Luo

  4. Department of Pharmacy, Qingdao Stomatological Hospital, No.17, Dexian Road, Shinan District, Qingdao, 266001, People’s Republic of China

    Fang Wang

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  1. Bing-Chang Xin
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Correspondence to Fang Wang.

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Figure. S1

The adipogenic differentiation of hDPSCs. Oil red-O staining analysis for the lipid droplets formation in the hDPSCs after 14 days induction by special adipogenic differentiation induction medium (LM) containing 10% FBS, 1 mg/ml insulin, 1 mM dexamethasone, 0.5 mM isobutyl methylxanthine and 100 mM indomethacin. Cells cultured with routine medium (RM) was considered as control. (PNG 371 kb)

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(TIF 974 kb)

Figure. S2

The effects of Berberine on the adipogenic differentiation of hDPSCs. hDPSCs were cultured with special adipogenic differentiation induction medium containing 1, 5 μM Berberine or 0.1% DMSO for 14 days, and Oil red-O staining was used to analyzed the lipid droplets formation. (PNG 542 kb)

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(TIF 1445 kb)

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Xin, BC., Wu, QS., Jin, S. et al. Berberine Promotes Osteogenic Differentiation of Human Dental Pulp Stem Cells Through Activating EGFR-MAPK-Runx2 Pathways. Pathol. Oncol. Res. 26, 1677–1685 (2020). https://doi.org/10.1007/s12253-019-00746-6

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  • DOI: https://doi.org/10.1007/s12253-019-00746-6

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