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Dental pulp stem cells overexpressing stromal-derived factor-1α and vascular endothelial growth factor in dental pulp regeneration

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Abstract

Objectives

The current study aimed to investigate the effects of vascular endothelial growth factor (VEGF) and stromal cell-derived factor-1α (SDF-1α) overexpressing dental pulp stem cells (DPSCs) in vascularized dental pulp regeneration in vivo.

Materials and methods

Human DPSCs were transfected with VEGF or SDF-1α using premade lentiviral particles. Overexpression was verified by quantitative polymerase chain reaction (q-PCR), enzyme-linked immunosorbent assay (ELISA), and western blot analysis. Effects of SDF-1α and VEGF overexpressing DPSCs on their proliferation (CCK-8 and MTT assays) and endothelial vascular-tube formation (Matrigel assay) were investigated in vitro. Human tooth roots sectioned into 6-mm segments were injected with gene-modified DPSCs encapsulated in PuraMatrix hydrogel and implanted in the dorsum of severe-combined-immunodeficient (SCID) mice. Implants were retrieved after 4 weeks and examined for regenerated pulp-like tissue and vascularization using histology and immunohistochemistry. p < 0.05 was considered statistically significant.

Results

Gene-modified DPSCs expressed significantly high levels (p < 0.05) of SDF-1α and VEGF mRNA and proteins, respectively. Transfected DPSCs showed a significantly higher cell proliferation compared to that of wild-type DPSCs. Furthermore, they enhanced endothelial cell migration and vascular-tube formation on Matrigel in vitro. When injected into tooth root canals and implanted in vivo, DPSCs/SDF-1α + DPSCs/VEGF-mixed group resulted in significantly increased length of regenerated pulp-like tissue within the root canals compared to that of wild-type DPSCs/VEGF and DPSCs/SDF-1α groups. Vessel area density was significantly higher in DPSCs/SDF-1α and mixed DPSCs/SDF-1α + DPSCs/VEGF groups than in DPSCs-VEGF alone or wild-type DPSCs groups.

Conclusion

A combination of VEGF-overexpressing and SDF-1α-overexpressing DPSCs could enhance the area of vascularized dental pulp regeneration in vivo.

Clinical relevance

Enhancing vascularization in pulp regeneration is crucial to overcome the clinical limitation of the limited blood supply to the root canals via a small apical foramen enclosed by hard dentin.

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Funding

The work was supported by the General Research Fund grants (project codes 17126914 and HKU 784912) awarded to Chengfei Zhang by the Research Grants Council of Hong Kong.

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Author notes

  1. Lifang Zhu and Waruna Lakmal Dissanayaka contributed equally to this work.

Authors and Affiliations

  1. Endodontology, Faculty of Dentistry, The University of Hong Kong, 3A15, Prince Philip Dental Hospital, 34, Hospital Road, Hong Kong, SAR, China

    Lifang Zhu & Chengfei Zhang

  2. Applied Oral Sciences, Faculty of Dentistry, The University of Hong Kong, Hong Kong, SAR, China

    Waruna Lakmal Dissanayaka

Authors
  1. Lifang Zhu
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  2. Waruna Lakmal Dissanayaka
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  3. Chengfei Zhang
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Correspondence to Chengfei Zhang.

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The authors declare that they have no conflict of interest.

Ethical approval

This article does not contain any studies with human participants by any of the authors. All applicable institutional guidelines set by the Committee on the Use of Live Animals in Teaching and Research (CULATR) of The University of Hong Kong for the care and use of animals were followed.

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For this type of study, formal consent is not required.

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Zhu, L., Dissanayaka, W.L. & Zhang, C. Dental pulp stem cells overexpressing stromal-derived factor-1α and vascular endothelial growth factor in dental pulp regeneration. Clin Oral Invest 23, 2497–2509 (2019). https://doi.org/10.1007/s00784-018-2699-0

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  • DOI: https://doi.org/10.1007/s00784-018-2699-0

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