Lithium-containing surface pre-reacted glass fillers enhance hDPSC functions and induce reparative dentin formation in a rat pulp capping model through activation of Wnt/β-catenin signaling

Surface pre-reacted glass (S-PRG) fillers are new bioactive molecules used in dental clinic work to fill tooth defects. These fillers release various types of ions (Al⁺³, BO^-3, Na⁺, SiO₃^-2, Sr⁺² and F^-) and exhibit high biocompatibility, antibacterial capability, reduced plaque accumulation, and enhanced osteoblast differentiation. We previously showed that cement of S-PRG fillers could induce tertiary dentin formation in rat models. Previous work also showed that lithium ions can activate the Wnt/β-catenin signaling pathway in vitro and induce dentin formation in pulpotomized teeth in vivo. In the current study, we sought to enhance the effect of S-PRG cement by incorporating LiCl. We show that treatment of human dental pulp stem cells with eluates from S-PRG/LiCl combination cements leads to an upregulation in cell migration, differentiation, and mineralization in vitro. In pulp-capping animal trials, we found that S-PRG/LiCl cements could induce tertiary dentin formation 28-days post-capping. At 7 days post-capping, we identified both β-catenin and Axin2 expression using immunofluorescence, indicative of Wnt/β-catenin signaling activity. In conclusion, S-PRG/LiCl cement is highly effective in promoting human dental pulp stem cells profiles and in enhancing reparative dentin formation in rat teeth through activation of the Wnt/β-catenin canonical signaling pathway. STATEMENT OF SIGNIFICANCE: This is the first study to assess the behavior of S-PRG fillers containing lithium ions on human dental pulp stem cells. We show that this new combination cement promotes positive cell responses by activating the endogenous Wnt/β-catenin signaling pathway in the pulp. The Wnt/β-catenin canonical signaling pathway is involved in many developmental and wound healing processes. The released lithium ions from the S-PRG cement were systematically detected <0.01 mmol/L in our rat model. But it was efficient to induce tertiary dentin formation at the defect site. Since this novel bioactive cement is potentially a promising material for clinical pulp regenerative therapy, future human clinical trials will be needed.