Shear stress promotes differentiation of stem cells from human exfoliated deciduous teeth into endothelial cells via the downstream pathway of VEGF-Notch signaling

Wang,Penglai; Zhu,Shaoyue; Yuan,Changyong; Wang,Lei; Xu,Jianguang; Liu,Zongxiang

doi:10.3892/ijmm.2018.3761

Shear stress promotes differentiation of stem cells from human exfoliated deciduous teeth into endothelial cells via the downstream pathway of VEGF-Notch signaling

Authors:
- Penglai Wang
- Shaoyue Zhu
- Changyong Yuan
- Lei Wang
- Jianguang Xu
- Zongxiang Liu
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Affiliations: Dental Implant Center, Xuzhou Stomatological Hospital, Xuzhou, Jiangsu, P.R. China, Department of Orthodontics, Xuzhou Stomatological Hospital, Xuzhou, Jiangsu, P.R. China, Department of Periodontics, Xuzhou Stomatological Hospital, Xuzhou, Jiangsu, P.R. China, The Discipline of Endodontology, Faculty of Dentistry, The University of Hong Kong, Hong Kong, SAR, P.R. China, Department of ExperDignosis, Xuzhou Stomatological Hospital, Xuzhou, Jiangsu, P.R. China
Published online on: July 6, 2018 https://doi.org/10.3892/ijmm.2018.3761
Pages: 1827-1836

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Abstract

Effects of shear stress on endotheliaxl differentiation of stem cells from human exfoliated deciduous teeth (SHEDs) were investigated. SHEDs were treated with shear stress, then reverse transcription-quantitative polymerase chain reaction (RT-qPCR) was performed to analyse the mRNA expression of arterial markers and western blot analysis was performed to analyse protein expression of angiogenic markers. Additionally, in vitro matrigel angiogenesis assay was performed to evaluate vascular-like structure formation. The secreted protein expression levels of the vascular endothelial growth factor (VEGF) of SHEDs after shear stress was also quantified using corresponding ELISA kits. Untreated SHEDs seeded on Matrigel cannot form vessel-like structures at any time points, whereas groups treated with shear stress formed a few vessel-like structures at 4, 8 and 12 h. When SHEDs were treated with EphrinB2-siRNA for 24, the capability of vessel-like structure formation was suppressed. After being treated with shear stress, the expression of VEGF, VEGFR2, DLL4, Notch1, EphrinB2, Hey1 and Hey2 (arterial markers) gene expression was significantly upregulated, moreover, the protein levels of VEGFR2, EphrinB2, CD31, Notch1, DLL4, Hey1, and Hey2 were also significantly upregulated. Both the mRNA and protein expression levels of EphB4 (venous marker) were downregulated. The average VEGF protein concentration in supernatants secreted by shear stress treated SHEDs groups increased significantly. In conclusion, shear stress was able to induce arterial endothelial differentiation of stem cells from human exfoliated deciduous teeth, and VEGF-DLL4/Notch‑EphrinB2 signaling was involved in this process.

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