hDPSC-derived Secretomes Enhance Odontogenic Differentiation and Suppress Inflammation: A Potential Cell-free Approach for Regenerative Endodontics

Introduction: Secretomes are bioactive molecules secreted by stem cells that contain growth factors, anti-inflammatory agents, and proteins involved in tissue repair. This study aimed to evaluate the anti-inflammatory and odontogenic effects of human dental pulp stem cells (hDPSCs)-derived secretomes. Methods: Secretomes were isolated from hDPSCs through stepwise filtration and centrifugation. Gene ontology analysis was performed to characterize the secretome composition. The effects of secretomes on cell migration were assessed via wound healing assay. Anti-inflammatory and odontogenic effects were evaluated using quantitative real-time polymerase chain reaction and western blotting for inflammatory markers and odontogenic differentiation markers. Alkaline phosphatase staining was performed to assess mineralization under lipopolysaccharide-induced inflammatory conditions. Data were statistically analyzed using one-way analysis of variance and Tukey's posthoc test (P < .05 was considered significant). Results: Gene ontology analysis confirmed key functional components of the secretome. The wound healing assay demonstrated that 100 μg/mL secretome treatment significantly enhanced human dental pulp cells migration. Secretome treatment significantly attenuated lipopolysaccharide-induced upregulation of inflammatory markers (P < .05) while upregulating odontoblastic differentiation markers and promoting mineralization (P < .05). Conclusion: hDPSC-derived secretomes exhibit anti-inflammatory and odontogenic effects, potentially mediated by MAPK signaling pathways. These findings suggest that secretomes may serve as a novel cell-free strategy for regenerative endodontics, offering therapeutic potential for vital pulp therapy and dentin-pulp regeneration.