Supporting data for Wnt targeted Therapeutics in Vital Pulp Therapy

Vital Pulp Therapy (VPT) increasingly focuses on regenerative repair using endogenous bioactive molecules, requiring understanding of pulp/dentine biology. Chapter 1 details pulp cells, signaling molecules, and ECM, providing insight into cell behavior and therapeutics.Targeting the Wnt pathway is key for hard tissue repair, meeting AAE/ESE biological criteria. While GSK3β inhibition activates Wnt, small-molecule inhibitors suffer from poor solubility, cytotoxicity, and <i>irreversible</i> inhibition, posing risks due to Wnt's diverse roles. This project addresses these limitations.This work comprehensively evaluates L803-mts, a novel <i>peptide-based</i>, substrate-competitive GSK3β inhibitor designed to overcome small-molecule drawbacks. Chapter 2 selected L803-mts over other peptides for its superior odontogenic differentiation of DPSCs (therapeutically key pulp cells).Wnt activation promotes dentine repair and sustains pulp cells (DPSCs, macrophages) under inflammation. DPSCs possess inherent anti-inflammatory/immunomodulatory activity. Chapter 3 details Wnt's modulation of NF-κB and DPSC-macrophage interactions during inflammation.This prompted investigating if L803-mts enhances these repair processes via Wnt activation under inflammation. Chapter 4 found that Wnt activation <i>magnitude</i> dictates outcomes: L803-mts (supra-physiological activation) promoted mineralization, while L803 (sub-threshold activation) enhanced self-renewal/slowed differentiation. Chapter 5 showed L803-mts boosts DPSC anti-inflammation.L803-mts excelled in differentiation and immunomodulation, ideal for pulp repair. It enhanced in vivo dentine repair, though less than MTA initially. Rapid mineralization (3 days) was offset by peptide instability (Chapter 6).Chapter 7 identified gelatin for dentine ECM mimicry and bioactive delivery. Chapter 8 engineered gelatin microspheres to load L803-mts, significantly improving stability. Incorporating microspheres into a hydrogel enabled sustained release. This system overcame prior limitations, achieving in vivo dentine repair efficacy comparable to MTA.