Wnt10a exhibit spatiotemporal singularity in the temporal changes of angiogenesis in regenerated pulp-like tissue

BACKGROUND: In tooth development, the Wnt/β-catenin pathway has been shown to play a crucial role in tooth germ formation and tooth differentiation. Regeneration can be considered a replay of development. Understanding and reproducing the involvement of Wnt10a-which is reported to be the causative gene for congenital multiple tooth absence and participates in tooth development with spatiotemporal specificity-in pulp regeneration is essential for realizing dental regenerative medicine. Therefore, this study was initiated based on the concept that evaluating the dynamics of Wnt10a during angiogenesis, which is thought to occur early in dental pulp regeneration, could contribute to a more detailed elucidation of the dental pulp regeneration mechanism. METHODS: Deciduous dental pulp stem cells (SHED) were isolated from human deciduous teeth, and conditioned medium (CM) was collected. In addition, after the induction of vascular differentiation of SHED, the temporal gene expression of Wnt10a, VEGF-A, Tie-2, and β-catenin was analyzed by q-PCR and protein expression by Western blotting and ELISA from 0 to 48 h and 3, 7, 14, and 21 days after the induction. RESULTS: Canonical Wnt signaling was activated during angiogenesis in regenerated pulp-like tissue induced by ectopic root grafting, and Wnt10a had spatio-temporal specificity. Tie-2 activation occurred during the process of induction of vascular differentiation in SHED. CONCLUSIONS: During angiogenesis in pulp regeneration, when SHED differentiate into blood vessels, Canonical Wnt signaling and VEGF-A are activated to form microvessels, and Tie-2 expression is enhanced to increase vessel circumference. Furthermore, Wnt10a was found to be activated in its early stages and decreased in its mature stages, with spatio-temporal specificity. Because its expression is very low, Wnt10a could be a biomarker to monitor the regenerative status of pulp regeneration.