Cell Contact Balances Mineralization And Angiogenesis In Dental Pulp Regeneration

This study investigates how intercellular contact dynamics between human dental pulp cells (hDPCs) and endothelial cells (HUVECs) coordinate tissue mineralization and vascularization during pulp regeneration. Tunable porous membrane cocultures (0.4μm vs. 1μm pores) and VitroGel hydrogel-based 3D subcutaneous transplants in nude mice were developed to modulate hDPCs/HUVECs interactions. Differentiation markers (ALP, DMP-1, DSPP, αSMA, NG-2, Bmi-1) and angiogenic activity (VE-Cadherin, CD105, VEGF, S1P) were analyzed via RT-qPCR, Western blot, CCK-8, Transwell, tubulogenesis assays, and histological staining. (1) Reduced cell contact (0.4μm pores) enhanced hDPC pericyte commitment (αSMA, NG-2, Bmi-1) and HUVECs vascular activity (VEGF, CD105, tubulogenesis). (2) Increased contact (1μm pores) promoted hDPCs odontogenic differentiation (ALP, DMP-1, DSPP) and vascular maturation (VE-Cadherin, S1P). (3) In vivo, low-contact cocultures showed elevated microvascular density (HE, VEGF), while high-contact groups exhibited active dentinogenesis (DMP-1, DSPP). Cell contact reciprocity dynamically balances hDPCs fate (pericyte vs. odontoblast) and HUVECs functionality (angiogenesis vs. stabilization). This bidirectional regulation provides a mechanistic foundation for optimizing vascularized pulp regeneration, addressing the critical challenge of synchronizing tissue mineralization and nutrient supply in regenerative dentistry. This study may provide evidence of contact-dependent crosstalk governing dual mineralization-angiogenesis outcomes, advancing strategies for functional dental tissue engineering.