Proteomic Profiling of PTEN Inhibition on Periodontal Ligament Stem Cells.

Phosphatase and tensin homolog (PTEN) is a critical regulator of cell proliferation, differentiation, and inflammatory balance. However, its downstream proteomic effects in periodontal ligament stem cells (PDLSCs) remain poorly understood. This study aimed to elucidate the proteomic alterations induced by PTEN inhibition and identify potential molecular pathways underlying periodontal regeneration. PDLSCs were treated with 5 μM VO-OHpic for 24 hours, followed by proteomic profiling using mass spectrometric analysis. The resulting proteomic data were analysed using Heatmapper, Metascape, and WebGestalt (WEB-based Gene Set Analysis Toolkit) databases, as well as Cytoscape, to evaluate protein expression patterns and protein-protein interaction networks. Proteomic analysis identified 7,525 proteins across all samples, with 3,497 proteins commonly expressed between groups. In the VO-OHpic-treated cells, proteins related to cytoskeletal dynamics (ACTR2, FNDC3A), vesicular transport (GGA1, RAB9A), and signal transduction (TRAF3IP3, GJB3) were upregulated. Enrichment analysis identified potential biological pathways related to membrane trafficking, secretion, and regulation of MAPK and Wnt signalling. Conversely, proteins linked to transcriptional and metabolic regulation (GLIS3, PHGDH, NADSYN1) and membrane trafficking (SNX1, REEP1) were downregulated. Cytokine signalling and interferon pathways were explored as the potential pathways involved in these downregulated proteins. In conclusion, PTEN inhibition by VO-OHpic was associated with proteomic changes in PDLSCs linked to regenerative phenotype, including the modulation of MAPK and Wnt signalling pathways alongside alterations in immune and metabolic pathways. These findings suggest that PTEN may function as a modulator balancing inflammatory regulation and differentiation processes, providing mechanistic insight into its potential role in periodontal tissue regeneration.