N-acetylcysteine restores osteogenesis in human periodontal ligament stem cells by attenuating HIF-1α-driven oxidative stress and glycolytic reprogramming

Authors

• Xiaochi Chang, Department of Periodontology, National Center of Stomatology & National Clinical Research Center for Oral Diseases, National Engineering Laboratory for Digital and Material Technology of Stomatology, Peking University School and Hospital of Stomatology, Beijing, China
• Yanghao Zhao, Department of Periodontology, National Center of Stomatology & National Clinical Research Center for Oral Diseases, National Engineering Laboratory for Digital and Material Technology of Stomatology, Peking University School and Hospital of Stomatology, Beijing, China
• Yang Wang, Department of Periodontology, National Center of Stomatology & National Clinical Research Center for Oral Diseases, National Engineering Laboratory for Digital and Material Technology of Stomatology, Peking University School and Hospital of Stomatology, Beijing, China
• Chang Shu, Department of Periodontology, National Center of Stomatology & National Clinical Research Center for Oral Diseases, National Engineering Laboratory for Digital and Material Technology of Stomatology, Peking University School and Hospital of Stomatology, Beijing, China
• Yi Feng, Department of Periodontology, National Center of Stomatology & National Clinical Research Center for Oral Diseases, National Engineering Laboratory for Digital and Material Technology of Stomatology, Peking University School and Hospital of Stomatology, Beijing, China
• Yichong Sun, Department of Periodontology, National Center of Stomatology & National Clinical Research Center for Oral Diseases, National Engineering Laboratory for Digital and Material Technology of Stomatology, Peking University School and Hospital of Stomatology, Beijing, China
• Guangxun Zhu, Department of Stomatology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei, China
• Jianxia Hou, Department of Periodontology, National Center of Stomatology & National Clinical Research Center for Oral Diseases, National Engineering Laboratory for Digital and Material Technology of Stomatology, Peking University School and Hospital of Stomatology, Beijing, ChinaFollow

First Page

1486

Last Page

1498

Abstract

Background/purpose: Periodontitis-associated hypoxia signaling and oxidative stress severely impair the regenerative capacity of human periodontal ligament stem cells (hPDLSCs). This study investigated whether antioxidant intervention rescues hPDLSC osteogenesis under CoCl₂-induced HIF-1α stabilization by modulating PI3K/Akt/HIF-1α/HO-1 signaling and glycolytic reprogramming.

Materials and methods: Primary hPDLSCs isolated from healthy premolars were characterized by flow cytometry and multilineage differentiation. To simulate periodontitis-associated stress, a CoCl₂-induced HIF-1α-stabilizing condition was established. Cells were treated with the antioxidant N-acetylcysteine (NAC), the glycolysis inhibitor 2-deoxy-D-glucose (2-DG), or the PI3K inhibitor LY294002. Redox balance (DCFH-DA, MDA, GPX activity), glycolysis-related markers (HK2, GLUT1, lactate), and osteogenic-related markers (RUNX2, ALP, OCN) were quantified via qRT-PCR, Western blot, and biochemical assays, alongside an analysis of the PI3K/Akt/HIF-1α/HO-1 signaling axis.

Results: CoCl₂ exposure triggered profound redox imbalance and significantly impaired hPDLSC osteogenesis. Crucially, NAC effectively reversed these damages and restored osteogenic capacity. Mechanistically, CoCl₂-induced HIF-1α stabilization activated PI3K/Akt/HO-1 signaling, a pathological cascade attenuated by either NAC or targeted PI3K inhibition. Furthermore, CoCl₂ forced a pronounced glycolytic shift characterized by upregulated HK2, GLUT1, and lactate production, which was directly intercepted by ROS clearance. Functionally, targeted glycolysis inhibition via 2-DG also successfully rescued the osteogenic potential of HIF-1α-stabilized hPDLSCs.

Conclusion: HIF-1α stabilization suppresses hPDLSC osteogenesis by inducing both oxidative stress and pathological glycolytic reprogramming. Targeted antioxidant intervention effectively mitigates these impairments, highlighting the ROS/PI3K/Akt/HIF-1α axis as a key link between redox imbalance and metabolic dysfunction, and suggesting a potential therapeutic target for periodontal regeneration.

Recommended Citation

Chang, Xiaochi; Zhao, Yanghao; Wang, Yang; Shu, Chang; Feng, Yi; Sun, Yichong; Zhu, Guangxun; and Hou, Jianxia (2026) "N-acetylcysteine restores osteogenesis in human periodontal ligament stem cells by attenuating HIF-1α-driven oxidative stress and glycolytic reprogramming," Journal of Dental Sciences: Vol. 21: Iss. 3, Article 17.
Available at: https://jds.ads.org.tw/journal/vol21/iss3/17

Publication Date

2026