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Abstract <p>Dental pulp stem cells (DPSCs) play a critical role in maintaining dental pulp homeostasis and supporting dentin-pulp complex regeneration, while tumor necrosis factor-α (TNF-α)-mediated inflammation severely impairs their biological functions. This study explored the role of the miR-335-5p/DKK1/autophagy axis in TNF-α-induced DPSCs dysfunction. Human DPSCs were stimulated with 20&#xa0;ng/mL TNF-α; miR-335-5p overexpression and DKK1 silencing were achieved via transfection. qRT-PCR, Western blot, SA-β-gal staining, immunofluorescence, transmission electron microscopy, ALP/ARS staining, and dual-luciferase assay were used to detect related indicators. Results showed TNF-α downregulated miR-335-5p, upregulated DKK1, inhibited autophagy, induced senescence, disrupted cytoskeleton, and suppressed osteogenesis; miR-335-5p directly targeted DKK1’s 3′-UTR. Overexpressing miR-335-5p or silencing DKK1 restored DPSCs’ autophagic flux, alleviated senescence, and rescued osteogenic potential. In conclusion, the miR-335-5p/DKK1/autophagy axis mediates TNF-α-induced DPSCs dysfunction, and targeting this axis may improve stem cell-based dental pulp and bone reconstruction under inflammation.</p>

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The miR-335-5p/DKK1/autophagy axis regulates TNF-α-mediated dysfunction of dental pulp stem cells

  • 期刊论文

  • 发表日期：

    2026年7月24日

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  • Guijuan Feng,
  • Lan Huang,
  • Xiaofeng Sun,
  • Fanting Sun,
  • Han Su,
  • Haoting Shi,
  • Ke Zheng,
  • Shiyu Shi

摘要

Abstract

Dental pulp stem cells (DPSCs) play a critical role in maintaining dental pulp homeostasis and supporting dentin-pulp complex regeneration, while tumor necrosis factor-α (TNF-α)-mediated inflammation severely impairs their biological functions. This study explored the role of the miR-335-5p/DKK1/autophagy axis in TNF-α-induced DPSCs dysfunction. Human DPSCs were stimulated with 20 ng/mL TNF-α; miR-335-5p overexpression and DKK1 silencing were achieved via transfection. qRT-PCR, Western blot, SA-β-gal staining, immunofluorescence, transmission electron microscopy, ALP/ARS staining, and dual-luciferase assay were used to detect related indicators. Results showed TNF-α downregulated miR-335-5p, upregulated DKK1, inhibited autophagy, induced senescence, disrupted cytoskeleton, and suppressed osteogenesis; miR-335-5p directly targeted DKK1’s 3′-UTR. Overexpressing miR-335-5p or silencing DKK1 restored DPSCs’ autophagic flux, alleviated senescence, and rescued osteogenic potential. In conclusion, the miR-335-5p/DKK1/autophagy axis mediates TNF-α-induced DPSCs dysfunction, and targeting this axis may improve stem cell-based dental pulp and bone reconstruction under inflammation.