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Heliyon. 2019 Apr 28;5(4):e01560. doi: 10.1016/j.heliyon.2019.e01560. eCollection 2019 Apr.

Angiogenic properties of dental pulp stem cells conditioned medium on endothelial cells in vitro and in rodent orthotopic dental pulp regeneration.

Author information

1
School of Dentistry, Centro Universitário das Faculdades Metropolitanas Unidas (FMU), São Paulo, SP, Brazil.
2
Departamento de Medicina, Divisão de Nefrologia, Universidade Federal de São Paulo, São Paulo, SP, Brazil.
3
Institute of Life Sciences, Universidade Federal de Juiz de Fora (UFJF), Governador Valadares, MG, Brazil.
4
School of Dentistry, Universidade Ibirapuera (UNIB), Sao Paulo, SP, Brazil.
5
Department of Restorative Dentistry and Endodontics, School of Dentistry, University of Sao Paulo (USP), Sao Paulo, SP, Brazil.
6
School of Medicine, UNINOVE University, São Paulo, São Paulo, Brazil.

Abstract

Objectives:

To evaluate the effect of SHED-CM on the proliferation, differentiation, migration ability, cell death, gene expression and production of VEGF of HUVEC in vitro and in a rodent orthotopic dental pulp regeneration.

Methods:

Three culture media [M199, DMEM/Ham's F12 and DMEM/Ham's F12 conditioned by SHEDs] were used as experimental groups. SHED-CM was prepared maintaining confluent cells in culture without serum for 3 days. The proliferation and cell death marker of HUVECs were assessed using flow cytometry. The capacity of formation of vascular-like structures was analyzed in cells grown over Matrigel® in hypoxic condition. HUVECs migration was followed using the scratch test. VEGF-A expression in HUVECs was assessed using real time RT-qPCR; and VEGF synthesis with ELISA test. SHED-CM was also applied in rodent ortotopic model of dental pulp regeneration in rats. The formed tissue was submitted to histological and immunohistochemical analyses.

Results:

SHED-CM promoted significantly lower expression of 7AAD in HUVECs; whereas the expression of the Ki67 was similar in all groups. The vascular-like structures were observed in all groups. Migration of SHED-CM group was faster than DMEM/Ham's F12. SHED-CM induced similar expression of VEGF-A than M199, and higher than DMEM/Ham's F12. SHED-CM induced significantly higher VEGF synthesis than other media. SHED-CM induced formation of a vascularized connective tissue inside the root canal.

Conclusion:

The study showed that SHEDs release angiogenic and cytoprotective factors, which are of great importance for tissue engineering.

Clinical significance:

SHED-CM could be an option to the use of stem cells in tissue engineering.

KEYWORDS:

Bioengineering; Cell biology

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