Osteogenic potential of Frondoside A in human periodontal ligament cells: an RNA-Seq analysis.

ABSTRACT

PURPOSE: The aim of this study was to evaluate the effects of Frondoside A (FA) on the osteogenic differentiation of human periodontal ligament (PDL) cells. METHODS: Human PDL cells were cultured in osteogenic medium and treated with FA at concentrations of 0, 0.05, and 0.2 µM for 14 days. The expression levels of genes associated with osteogenic differentiation were assessed using quantitative real-time polymerase chain reaction analysis. Subsequently, RNA sequencing was performed to identify enriched gene sets following FA treatment. Alkaline phosphatase (ALP) activity was measured to confirm the osteogenic potential of FA. RESULTS: Treatment with 0.2 µM FA significantly increased the expression levels of runt-related transcription factor 2 (RUNX2), ALP, and osteocalcin (OCN) at day 3, while also significantly elevating the expression of dentin sialophosphoprotein (DSPP), RUNX2, ALP, OCN, and osterix (OSX) at day 14 (P<0.017). Hallmark gene sets enriched during FA treatment were associated with the KRAS (normalized enrichment score [NES]=2.02, Q=0.000), interferon alpha (IFN-α) (NES=1.88, Q=0.001), IFN-γ (NES=1.85, Q<0.001), hypoxia (NES=1.79, Q=0.001), and p53 (NES=1.77, Q=0.001) signaling pathways. Additionally, treatment with 0.2 µM FA significantly intensified ALP staining at day 14 (P<0.05). CONCLUSIONS: Within the limitations of this study, FA treatment influenced periodontal regeneration by promoting the osteogenic differentiation of human PDL cells.