Forkhead Box O1 Promotes Osteogenesis of Periodontal Ligament Stem Cells Via Glycolysis-Related Metabolic Reprogramming

  • Int Dent J. 2026 Aug;76(4):109664. doi: 10.1016/j.identj.2026.109664.
Jiaxin Zou  1 ,  Jiang Guo  1 ,  Qian Zeng  1 ,  Zhuoyao Liang  1 ,  Luchen Gui  1 ,  Xiaojun Huang  2 ,  Jiacheng Lin  3
Affiliations
  • 1. Guangdong Provincial Key Laboratory of Stomatology, Guanghua School of Stomatology, Hospital of Stomatology, Sun Yat-Sen University, Guangzhou, People's Republic of China.
  • 2. Guangdong Provincial Key Laboratory of Stomatology, Guanghua School of Stomatology, Hospital of Stomatology, Sun Yat-Sen University, Guangzhou, People's Republic of China. Electronic address: [email protected].
  • 3. Guangdong Provincial Key Laboratory of Stomatology, Guanghua School of Stomatology, Hospital of Stomatology, Sun Yat-Sen University, Guangzhou, People's Republic of China. Electronic address: [email protected].
PMID: 42247869 DOI: 10.1016/j.identj.2026.109664
Abstract

Introduction and aims: Predictable periodontal regeneration is limited by variability in the osteogenic capacity of candidate cell sources. This study aimed to investigate whether forkhead box O1 (FOXO1) enhances the osteogenic differentiation of periodontal ligament stem cells (PDLSCs) via glycolysis-related metabolic reprogramming.

Methods: PDLSCs stably overexpressing FOXO1 were established, and osteogenic differentiation was evaluated by analysing osteogenic marker expression, ALP staining intensity and extracellular matrix mineralization. RNA Sequencing and gene-set enrichment analyses were performed to identify FoxO1-related transcriptional programs. Glycolytic activity was assessed by measuring glucose levels, lactate production, 2-NBDG glucose uptake and the expression of key glycolytic Enzymes. The contribution of glycolysis-related metabolism to FoxO1-driven osteogenic differentiation was examined using rotenone and dichloroacetate (DCA). FOXO1 activity was pharmacologically inhibited using AS1842856.

Results: FOXO1 overexpression significantly enhanced osteogenic differentiation of PDLSCs, as evidenced by increased osteogenic marker expression, ALP staining intensity and matrix mineralization (P < .05). Transcriptomic profiling revealed significant enrichment of glycolysis-related gene sets in FoxO1-overexpressing PDLSCs (P < .05). Functionally, FOXO1 reduced glucose levels, increased lactate production and 2-NBDG uptake, and upregulated key glycolytic Enzymes at the mRNA and protein levels (P < .05). FOXO1 inhibition partially attenuated these glycolysis-related changes and reduced RUNX2 and OCN expression during osteogenic induction. Rotenone-induced glycolytic shift further enhanced the osteogenic differentiation of PDLSCs (P < .05). Notably, DCA treatment attenuated the FoxO1-driven osteogenic enhancement (P < .05).

Conclusion: These findings identify FOXO1 as a metabolic regulator that promotes osteogenic differentiation of PDLSCs through glycolysis-related metabolic reprogramming.

Clinical relevance: Targeting FoxO1-regulated glycolytic metabolism may represent a novel strategy to enhance stem cell-based periodontal regeneration.

Keywords
FoxO1; Glycolysis; Osteogenic differentiation; Periodontal ligament stem cells.
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