Neoeriocitrin Targeting Beclin1 Deubiquitination and Autophagy in Osteogenic Differentiation of Human Dental Pulp Stem Cells

Abstract Human dental pulp stem cells (hDPSCs) are dental‐derived mesenchymal stem cells with robust multipotent differentiation potentials, rendering them promising for bone tissue engineering. However, their differentiation relies on expensive, hard‐to‐control growth factors. Neoeriocitrin (Neo), a natural flavonoid, promotes cell proliferation and regulates alkaline phosphatase activities. However, Neo's effect on hDPSCs osteogenesis and bone regeneration is unknown. This study investigated Neo's impact on hDPSCs osteogenic differentiation and its mechanisms for bone regeneration. Neo effectively boosted hDPSCs osteogenic differentiation in vitro and facilitated bone regeneration in rat calvarial defects in vivo. Thermal proteome profiling revealed Neo directly binds Beclin1, validated by cellular thermal shift assay, molecular docking, and molecular dynamics. Neo stabilized Beclin1 by inhibiting ubiquitination‐mediated degradation, increasing autophagy in Neo‐treated hDPSCs. Furthermore, Neo‐enhanced osteogenic differentiation is activated by the Beclin1 network, pivotal for bone regeneration. Elucidating the Neo‐Beclin1 interaction provides insights into regulating hDPSCs differentiation and opens new avenues for enhancing bone regeneration strategies.