CRT Silencing Inhibits ECM Synthesis in HGFs Cultured on Three-Dimensional PLGA Scaffolds Via Inhibiting The CaN/NFAT3 Signalling Pathway

The stability of orthodontic treatment is believed to be greatly affected by the compression and contraction of gum tissue, but the underlying molecular mechanism remains unclear. The aim of the current study was to explore the effects of mechanical forces on CRT, CaN, NFAT3, p-NFAT3 and COL-I expression in human gingival fibroblasts (HGFs) cultured on three-dimensional (3D) poly(lactic-co-glycolic acid) (PLGA) scaffolds. A mechanical force of 25 g/cm 2 was applied to HGFs for 0, 6, 24, 48, and 72h. The expression of CRT, CaN, NFAT3, p-NFAT3, and COL-I were examined by reverse transcription-quantitative polymerase chain reaction and western blotting analysis. The application of mechanical force on HGFs cultured on the 3D PLGA scaffolds led to a significant increase in CRT, CaN, and COL‑I expression, as well as a reduction in p-NFAT3 expression. The mechanical force effects were reversed by silencing CRT. By lowering the CRT expression, p-NFAT3 was upregulated, while CaN and COL‑I were downregulated in HGFs. These findings suggested that downregulation of CRT inhibited extracellular matrix synthesis, potentially via CaN/NFAT3 signalling pathway. Therefore, CRT may serve as a potential therapeutic target against gingival fibrosis.