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

• Published in: Heliyon Vol. 5; no. 4; p. e01560
• Main Authors: de Cara, Sueli Patricia Harumi Miyagi, Origassa, Clarice Silvia Taemi, de Sá Silva, Fernando, Moreira, Maria Stella N.A., de Almeida, Danilo Candido, Pedroni, Ana Clara Fagundes, Carvalho, Giovanna Lopes, Cury, Diego Pulzatto, Câmara, Niels Olsen Saraiva, Marques, Márcia Martins
• Format: Journal Article
• Language: English
• Published: England Elsevier Ltd 01.04.2019; Elsevier
• Subjects: Bioengineering; Cell biology; Bioengineering; Cell biology
• ISSN: 2405-8440; 2405-8440
• DOI: 10.1016/j.heliyon.2019.e01560

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. 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. 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. The study showed that SHEDs release angiogenic and cytoprotective factors, which are of great importance for tissue engineering. SHED-CM could be an option to the use of stem cells in tissue engineering.