Application of dynamic hydrogels for dental pulp regeneration: a comparative study on cellular behavior and inflammatory modulation

Abstract

Introduction Regenerative endodontic therapy aims to restore the pulp-dentin complex through biological approaches; however, current cell-free techniques often fail to achieve true dental pulp regeneration and may result in fibrous tissue formation or canal calcification. This study investigated dynamic hydrogels as advanced scaffolds for dental pulp stem cell (DPSC)-mediated regeneration, leveraging their viscoelastic properties to regulate cellular behavior and immune responses. Methods Dynamic hydrogels were synthesized using host-guest chemistry (Gel-Mal/HA-Ada-CD-SH) to mimic aspects of the native extracellular matrix and were compared with non-dynamic Gel-Mal hydrogels. DPSCs and macrophages were encapsulated within the hydrogels to evaluate cellular viability, morphology, inflammatory marker expression, and gene expression profiles. Results Dynamic hydrogels exhibited a distinct porous microarchitecture and significantly increased NF-κB and VEGF expression while reducing IL-1RA expression compared with non-dynamic hydrogels. Gene expression analysis demonstrated significantly increased TGFβ1, SMAD3, and MAPK expression, together with reduced CD80 expression, in macrophages cultured within dynamic hydrogels. Although iNOS expression was lower in the dynamic hydrogel group, the difference was not statistically significant. Discussion These findings suggest that dynamic hydrogels do not simply suppress inflammatory signaling but may modulate inflammatory and tissue remodeling pathways while promoting angiogenesis-associated responses. Limitations of the study include the short-term in vitro design and evaluation of a limited panel of inflammatory and fibrosis-associated markers. Within these limitations, dynamic hydrogels demonstrated favorable cellular and immunomodulatory responses and may provide a biologically active microenvironment that supports cellular activities relevant to regenerative endodontic applications. Further in vivo studies are required to validate their translational potential.