The Role of Wharton’s Jelly Secretome in Promoting Dentin-pulp Complex Regeneration — An In Vitro Study on Human Dental Pulp Stem Cells

Introduction Regeneration of the dentin–pulp complex is hindered by its low-compliance anatomy. Wharton’s Jelly Secretome (WJS) offers a cell-free, minimally immunogenic approach with potential regenerative benefits. Materials and Methods Human dental pulp stem cells (hDPSCs; P3–P4) were cultured under osteogenic, angiogenic, or inflammatory (LPS-treated) conditions. WJS (12.5%, 25%, 50%) was applied. TNF-α, VEGF-A, and DSPP levels were quantified by ELISA. Mineralization in osteogenic cultures was assessed by Alizarin red staining at 21 days. The quantitative data were analyzed using one-way ANOVA at a significance level of 95% ( p <0.05), followed by LSD post hoc analysis for homogeneous data. Results WJS significantly reduced TNF-α ( p < 0.05), increased VEGF-A and DSPP expression ( p < 0.05), and enhanced mineral deposition in a concentration-dependent manner. Discussion WJS attenuated inflammation by reducing TNF-α in all LPS-stimulated hDPSC groups, with the lowest level at 12.5% early on, indicating suppression of a key cytokine involved in irreversible pulpitis. It also supported regeneration by increasing VEGF-A levels, particularly on day 7 (highest at 50%), suggesting enhanced angiogenesis and tissue homeostasis. In parallel, odontogenic/osteogenic differentiation progressed as reflected by increased DSPP (highest at 12.5% on day 7 and peaking at 50% on day 14) and the greatest mineralized nodule formation at 50% by day 21. Conclusion WJS demonstrates anti-inflammatory, pro-angiogenic, and pro-odontogenic effects on hDPSCs, supporting dentin–pulp regeneration. These findings position WJS as a promising cell-free therapeutic for regenerative endodontics, warranting further translational studies.