Influence of METTL3 knockdown on PDLSC osteogenesis in E. coli LPS-induced inflammation

Oral Dis. 2024 Jul;30(5):3225-3238. doi: 10.1111/odi.14763. Epub 2023 Oct 9.

Abstract

Objective: This study aimed to investigate the effect of METTL3 knockdown on osteogenic differentiation of human periodontal ligament stem cells (PDLSCs) in the weak inflammation microenvironments, as well as the underlying mechanisms.

Materials and methods: PDLSCs were stimulated by lipopolysaccharide from Escherichia coli (E. coli LPS), followed by quantification of METTL3. METTL3 expression was assessed using RT-qPCR and Western blot analysis in periodontitis. METTL3 knockdown PDLSCs were stimulated with or without E. coli LPS. The evaluation included proinflammatory cytokines, osteogenic markers, ALP activity, and mineralized nodules. Bioinformatics analysis and Western blot determined the association between METTL3 and the PI3K/Akt pathway.

Results: METTL3 was overexpressed in periodontitis. METTL3 knockdown in PDLSCs reduced proinflammatory cytokines, osteogenic markers, ALP activity, and mineralized nodules in both environments. Bioinformatics analysis suggested a link between METTL3 and the PI3K/Akt pathway. METTL3 knockdown inhibited PI3K/Akt signaling pathway activation.

Conclusion: METTL3 knockdown might inhibit osteogenesis in PDLSCs through the inactivation of PI3K/Akt signaling pathway. Concomitant findings might shed novel light on the roles and potential mechanisms of METTL3 in the LPS-stimulated inflammatory microenvironments of PDLSCs.

Keywords: METTL3; PI3K/Akt signaling pathway; bioinformatics; osteogenesis; periodontitis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Differentiation
  • Cells, Cultured
  • Cytokines / metabolism
  • Escherichia coli*
  • Female
  • Gene Knockdown Techniques*
  • Humans
  • Inflammation / metabolism
  • Lipopolysaccharides*
  • Male
  • Methyltransferases* / genetics
  • Methyltransferases* / metabolism
  • Osteogenesis* / genetics
  • Periodontal Ligament* / cytology
  • Periodontal Ligament* / metabolism
  • Periodontitis / genetics
  • Periodontitis / metabolism
  • Phosphatidylinositol 3-Kinases* / metabolism
  • Proto-Oncogene Proteins c-akt* / metabolism
  • Signal Transduction*
  • Stem Cells / metabolism

Substances

  • Lipopolysaccharides
  • Methyltransferases
  • METTL3 protein, human
  • Phosphatidylinositol 3-Kinases
  • Proto-Oncogene Proteins c-akt
  • Cytokines