Mechanosensitive PIEZO1 Regulates Angiogenic Signaling in Human Dental Pulp Stem Cells

Dental pulp stem cells (DPSCs) possess multilineage differentiation potential and contribute to dental pulp regeneration through odontoblast replacement and neovascularization. Although mechanosensitive ion channels such as PIEZO1 are known to regulate stem cell fate, their role in DPSC-mediated angiogenesis under mechanical stress remains poorly understood. In this study, we investigated the involvement of PIEZO1 in regulating angiogenic gene expression and endothelial differentiation of immortalized human DPSCs (hDPSC-K4DT). Compressive force was applied using an in vitro weight-loading model previously developed to mimic the mechanical stress present in the pulp microenvironment. RT-qPCR analysis revealed that VEGFA expression was significantly upregulated following pressure loading. Additionally, activation of PIEZO1 using Yoda1, a specific chemical agonist, further enhanced the expression of VEGFA, HIF-1α, and bFGF, without affecting markers of hard tissue differentiation. Conversely, siRNA-mediated knockdown of the mechanosensitive ion channel PIEZO1 attenuated the upregulation of angiogenesis-related genes and inhibited the formation of capillary-like structures in a 3D spheroid sprouting assay. These results indicate that PIEZO1 acts as a mechanosensor mediating angiogenic signaling in DPSCs, thereby promoting endothelial differentiation and capillary morphogenesis. Mechanical stimuli such as compressive force, occlusal loading, or orthodontic pressure may activate PIEZO1 in pulp-resident stem cells and thereby promote vascular regeneration. In conclusion, PIEZO1 functions as a key mechanotransduction channel that regulates angiogenesis-related gene expression and vascular structure formation in DPSCs. Modulating PIEZO1 activity could enhance the angiogenic capacity of DPSCs, offering a promising strategy for vascularized dental pulp regeneration in regenerative endodontics.