Erk1/2-Nf-Κb Crosstalk Mediates Mechanoregulation Of Dpscs

To elucidate how dynamic hydrostatic pressure regulates dental pulp stem cells(DPSCs) behavior and the response mechanisms of ERK1/2 and NF-κB signaling pathways. This project used a cell pressure loading device to simulate occlusal trauma.DPSCs(P3-P5) obtained via the method of tissue block combined with enzyme digestion were subjected to dynamic hydrostatic pressure(0-90 kPa, 0.1 Hz). CCK-8 and the cell scratch assay method were used to detected the proliferation and migration of DPSCs. Using qPCR and Western Blotting to detect the mRNA and protein expression of osteogenic/odontoblastic differentiation.Screening differentially expressed genes and enriched pathways through RNA sequencing.The expression of related factors and activation of pathways were further verified by qPCR and Western Blotting. Dynamic hydrostatic pressure(0-90kPa) inhibited the proliferation, migration,and the osteogenic/odontoblastic differentiation of DPSCs. RNA-seq identified differentially expressed genes linked to inflammation and the genes were further verified by qPCR and Western Blotting. Dynamic hydrostatic pressure activated ERK1/2 and inhibited NF-κB phosphorylation, after adding the ERK1/2 inhibitor, the expression of inflammatory can be inhibited and the inhibited osteogenic/odontoblastic differentiation can be stimulated. Dynamic hydrostatic pressure(0-90kPa) inhibited DPSCs proliferation, migration, and osteogenic/odontoblastic differentiation while inducing inflammatory activation.The study deciphers ERK1/2 and NF-κB crosstalk as a key mechanotransduction switch in DPSCs, suggesting a mechanobiological strategy for overcoming pressure-induced regeneration barriers.