Restoring Cell-Cell Junctions in DSPP-Deficient Odontoblasts Through Nanofibrous Topography and Wnt5a-Cdc42...

Biomaterial-based strategies offer therapeutic potential for inherited disorders such as Dentinogenesis imperfecta type II (DGI-II). DGI-II, a hereditary dental disorder caused by dentine sialophosphoprotein (Dspp) gene mutations, results in fragile, discoloured teeth susceptible to wear and decay. This study investigates the structural alterations in the dentine matrix induced by mutant DSPP and the underlying molecular mechanisms. Furthermore, we evaluated a therapeutic strategy combining nanofibrous substrates with molecular activation of the Wnt5a-Cdc42 pathway, aiming to mitigate these structural defects and restore dentine integrity.

A maxillary first premolar extracted from a patient with dentinogenesis imperfecta type II (DGI-II) carrying a known DSPP frameshift mutation was analysed using micro-computed tomography and scanning electron microscopy. MDPC-23 mouse dental pulp pre-odontoblasts were cultured in vitro on electrospun nanofibrous scaffolds with tunable fibre diameters. Dspp expression was silenced to investigate its regulatory role in cell-cell contact formation, while the Wnt5a-Cdc42 pathway was activated via recombinant Wnt5a protein and a small molecule. Data were analysed using Student's t-test or one-way ANOVA with a significance threshold of p < 0.05.

Decalcified or freeze-fractured samples exhibited thick fibres, sparse dentinal tubules, and a disorganised, non-layered dentine matrix, indicating that disrupted cell-cell junctions impair directional appositional dentine formation. Silencing Dspp expression in dental pulp cells led to reduced expression of junctional proteins, Zona occludens-1 (ZO-1) and Connexin43 (Cx43), which are essential for vectorial matrix apposition. Modulating fibre thickness and activating the Wnt5a-Cdc42 signalling axis restored both the expression and cellular localisation of ZO-1 and Cx43, thereby re-establishing cell-cell junctions and paracellular permeability in Dspp-silenced dental pulp cells.

The results provide evidence that Dspp contributes to the structural integrity of dentine by modulating ZO-1 and Cx43 expression and suggest that biomaterial and molecular interventions may offer supportive strategies to restore dentine structure and function in DGI-affected teeth, though they do not address the underlying genetic defect.