HK2-mediated Glycolysis Inhibits Mineralization of Cementoblasts Under Compression by Suppressing the Piezo1/Wnt Signaling

Background: Orthodontically induced inflammatory root resorption (OIIRR) is a prevalent and severe complication during orthodontic tooth movement (OTM).Glycolysis plays a crucial role in the inflammatory responses.This study aimed to improve the cell compression model and investigate whether Hexokinase 2 (HK2)-mediated glycolysis regulates cementoblasts' mineralization through the mechanosensitive Piezo1/Wnt signaling under compressive force.Methods: Mouse cementoblasts (OCCM-30) were cultured under compressive force with different buffer membranes to mimic the periodontal membrane.The flow cytometry and CCK-8 assay were utilized to evaluate cell apoptosis and viability.Piezo1 and HK2 were knocked down by small interfering RNA (siRNA).The level of Wnt/β-catenin signaling was detected by qRT-PCR and Western blotting, and the cellular localization of β-catenin was detected by immunofluorescence staining.Results: The viability and apoptosis of cementoblasts showed no significant change under compression at 2.0 g/cm 2 for 12 hours with Polytetrafluoroethylene (PTFE) buffer membrane.HK2-mediated glycolysis was increased in compressed cementoblasts with elevated ratio of the receptor activator of nuclear factor kappa-B ligand/osteoprotegerin (RANKL/OPG) and decreased expression of Piezo1 and mineralization-related markers.The Piezo1 activated Wnt signaling by increasing the nuclear translocation of β-catenin, which increased the levels of mineral-related markers.Whereas, knockdown of Piezo1 showed the opposite trend.Knockdown of HK2 to inhibit glycolysis partially reversed the compression-induced decline in Piezo1 and mineralization-related markers, as well as the rise in the RANKL/OPG ratio.Conclusions: The cell compression model with PTFE buffer membrane effectively reduced cell damage.HK2-mediated glycolysis inhibited mineralization and enhanced osteoclast induction in cementoblasts under compression by suppressing the mechanosensitive Piezo1/Wnt signaling.