Effects of Smad4 on the expression of caspase‑3 and Bcl‑2 in human gingival fibroblasts cultured on 3D PLGA scaffolds induced by compressive force

Abstract

Human gingival fibroblasts (HGFs) are the main cells that comprise gingival tissue, where they transfer mechanical signals under physiological and pathological conditions. The exact mechanism underlying gingival tissue reconstruction under compressive forces remains unclear. The present study aimed to explore the effects of Smad4, caspase-3 and Bcl-2 on the proliferation of HGFs induced by compressive force. HGFs were cultured on poly(lactide-co-glycolide) (PLGA) scaffolds under an optimal compressive force of 25 g/cm 2 . Cell viability was determined via Cell Counting Kit-8 assays at 0, 12, 24, 48 and 72 h. The expression levels of Smad4, caspase-3 and Bcl-2 were measured via reverse transcription-quantitative PCR and western blotting. The application of compressive force on HGFs for 24 h resulted in a significant increase in cell proliferation and Bcl-2 expression, but a significant decrease in the expression of Smad4 and caspase-3; however, inverse trends were observed by 72 h. Subsequently, a lentivirus was used to overexpress Smad4 in HGFs, which attenuated the effects of compressive force on HGF proliferation and Bcl-2 expression, but enhanced caspase-3 expression, suggesting that Smad4 may regulate compressive force-induced apoptosis in HGFs. In conclusion, these findings increased understanding regarding the mechanisms of compressive force-induced HGF proliferation and apoptosis, which may provide further insight for improving the efficacy and stability of orthodontic treatment.

Inhibition of circular RNA circ_0138959 alleviates pyroptosis of human gingival fibroblasts via the microRNA-527/caspase-5 axis

“…The short hairpin RNA (shRNA) lentivirus of circ_0138959, negative control (sh-nc) lentivirus, overexpressed caspase-5 (CASP5) lentiviral vector (len-CASP5), and negative control (len-nc) were constructed by GenePharma Co., Ltd. (Jiangsu, China). According to previous studies [ 19 , 20 ], the HGFs of each group were transfected with the specified vector and/or sequence using Lipofectamine 3000 (Invitrogen, California, USA) reagent for 48 h when 60%–70% convergence was achieved.…”

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