Acellular dental pulp and allogeneic cells for strategic pulp regeneration

Regenerative endodontics aims to restore the damaged pulp-dentin complex, moving beyond revascularization to functional tissue regeneration, which includes innervation. While dental pulp stem cells (DPSCs) have been widely studied, their clinical application is hindered by low cell yield and reduced regenerative capacity in late passages. This study investigates the feasibility of using Wharton's Jelly mesenchymal stem cells (WJ-MSCs) as an alternative allogenic cell source, combining them with a decellularized human dental pulp (dDP) scaffold as a platform crucial for functional pulp regeneration. Dental pulp tissues were successfully decellularized, confirmed by DNA and hydroxyproline quantification, and histological and SEM analyses showing robust preservation of collagen types I and III with efficient cell removal. WJ-MSCs were then successfully repopulated onto the dDP scaffolds, demonstrating cellular attachment and migration into the ECM. Repopulated dDP constructs exhibited a propensity for osteogenic and odontogenic differentiation, evidenced by Alizarin Red staining for mineral deposition and upregulated gene expressions of osteogenic (DLX5, OSTERIX) and odontogenic (DSPP) markers. Furthermore, neurogenic marker NCAM was upregulated, whereas inhibitory genes (TWIST2, MSX1), angiogenic markers (VE-CADHERIN, VEGF), and immune regulatory marker IDO1 were downregulated following differentiation. Overall, the findings suggest the feasibility of combining WJ-MSCs with dDP scaffolds as a novel approach for functional pulp regeneration, offering a promising pathway for future clinical translation in regenerative endodontics.