Stomatology ›› 2025, Vol. 45 ›› Issue (7): 529-539.doi: 10.13591/j.cnki.kqyx.2025.07.009

• Basic and Clinical Research • Previous Articles     Next Articles

Study on the promotion of osteo/odontogenic differentiation on SCAPs and anti-inflammatory effects of EGCG/CeO2-loaded nanoparticles in an inflammatory microenvironment

WU Yuting1,2,3, DAI Mingrui1,2,3, LENG Diya1, ZHU Tingting1, WU Tao1,2,3, WU Daming1,2,3()   

  1. Department of Endodontics, The Affiliated Stomatological Hospital of Nanjing Medical University, Nanjing 210029, China
  • Received:2025-02-05 Online:2025-07-28 Published:2025-07-24

Abstract:

Objective To investigate the effects of composite nanoparticles E/Ce@MCSNs loaded with epigallocatechin-3-gallate (EGCG) and cerium dioxide (CeO2) on the odonto/osteogenic differentiation of human stem cells from the apical papilla (SCAPs) and macrophage polarization under inflammatory conditions. Methods E/Ce@MCSNs were synthesized and characterized. Cell viability of SCAPs and RAW 264.7 cells treated with varying concentrations of E/Ce@MCSNs was assessed via CCK-8 assay. The antioxidant enzyme-mimetic activity of E/Ce@MCSNs was evaluated. Under simulated inflammatory conditions, intracellular reactive oxygen species (ROS) scavenging capacity was measured via DCFH-DA fluorescent probe. Alkaline phosphatase (ALP) staining/activity, alizarin red staining/semi-quantitative analysis, and RT-qPCR were performed to detect odonto/osteogenic differentiation markers, including dentin sialoprotein (DSPP), ALP, runt-related transcription factor 2 (Runx-2), type Ⅰ collagen (COL-Ⅰ), and osteopontin (OPN) in SCAPs. The effects of E/Ce@MCSNs on the odontogenic/osteogenic differentiation of SCAPs under this condition were evaluated. RT-qPCR were used to analyze cytokine expression (TNF-α, IL-1β, IL-6, iNOS, TGF-β, IL-10) and secreted TNF-α/IL-6 levels in RAW264.7 cells. The concentrations of TNF-α and IL-6 in cell culture supernatants were measured by ELISA. Results E/Ce@MCSNs exhibited excellent biocompatibility at concentrations ≤100 μg/mL. They demonstrated potent ROS-scavenging activity (P<0.05) and significantly enhanced ALP activity (P<0.001), promoted calcium nodule formation (P<0.001), and upregulated odonto/osteogenic gene expression (DSPP, ALP, Runx-2, COL-Ⅰ, OPN) in SCAPs under inflammatory conditions (P<0.05). In RAW264.7 cells, E/Ce@MCSNs downregulated pro-inflammatory cytokines (TNF-α, IL-1β, IL-6, iNOS) (P<0.01) and upregulated anti-inflammatory factors (TGF-β, IL-10)(P<0.001), while reducing TNF-α and IL-6 secretion (P<0.001). Conclusion E/Ce@MCSNs exert antioxidant and anti-inflammatory effects in apical periodontitis by scavenging excessive ROS, thereby promoting odonto/osteogenic differentiation of SCAPs.

Key words: reactive oxygen species, antioxidant, apical papilla stem cells, anti-inflammatory, osteogenic differentiation

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