Abstract
Objectives
The primary objective of this study was to explore the effects of hypoxic preconditioning on stem cells derived from human exfoliated deciduous teeth (SHED) and to identify key molecular regulators that facilitate vascularized differentiation. The research question addressed was how hypoxic conditions influence SHEDs to induce angiogenesis and whether specific molecular targets can be manipulated to enhance this process.
Materials and Methods
SHEDs were subjected to hypoxic preconditioning, and their behavior was assessed under controlled conditions. Cellular responses were evaluated through whole exome sequencing, which led to the identification of Eps15 homology domain-containing protein 1 (EHD1) as a significantly altered gene. The role of EHD1 was investigated through a series of in vitro and in vivo experiments, including post-transcriptional modification analysis, assessment of cytoskeletal protein regulation, measurement of VEGF secretion and downstream pathway investigation.
Results
Hypoxic preconditioning induced significant changes in SHEDs, particularly in the expression of EHD1. When the expression of EHD1 was inhibited, SHEDs were able to differentiate into larger vascular structures, with upregulated expression of vimentin and increased expression of VEGF and its receptors. This secretion, in turn, affects the recruitment of endothelial cells and influences angiogenesis.
Conclusions
This study demonstrates that EHD1 is a pivotal factor in the hypoxic preconditioning of SHEDs, playing a key role in the regulation of cytoskeletal proteins and influencing cytoskeletal and membrane behavior. By controlling the paracrine secretion of VEGF, EHD1 affects the recruitment of endothelial cells and angiogenesis.