EphrinB2 Regulates DPSC Pericyte-like Functions via the Focal Adhesion Pathway

Area of Science:

• Vascular biology
• Stem cell biology
• Tissue engineering

Background:

• Mural cells, like pericytes, are crucial for vascular development and stabilization.
• Dental pulp stem cells (DPSCs) interact with pericytes and are regulated by EphB4/ephrinB2 signaling.
• The precise role of EphB4/ephrinB2 in DPSC pericyte function and EC interaction is unclear.

Purpose of the Study:

• To elucidate how EphB4/ephrinB2 signaling influences DPSC pericyte biology and their interactions with endothelial cells (ECs).
• To investigate the molecular mechanisms underlying ephrinB2-mediated regulation of DPSC-EC interactions.

Main Methods:

• Single-cell RNA sequencing and immunofluorescence staining of human dental pulp.
• Bulk RNA-sequencing to compare DPSCs with brain vascular pericytes.
• In vitro coculture of DPSCs and ECs, including EFNB2 knockdown and overexpression.
• 3D fibrin bead assay and focal adhesion protein assessment.

Main Results:

• DPSCs exhibit transcriptomic similarity to vascular pericytes.
• EFNB2 modulation in DPSCs altered proliferation, adhesion, migration, and contractility.
• EFNB2 overexpression enhanced EC sprouting and pericyte coverage in 3D models.
• These effects are mediated by the Src/FAK/paxillin signaling pathway.

Conclusions:

• EphrinB2 signaling critically regulates DPSC pericyte functions and EC adhesion via the Src/FAK/paxillin pathway.
• Targeting ephrinB2 signaling is a potential strategy for enhancing vascular formation in dental pulp tissue engineering.