Abstract
OBJECTIVE: To assess OPG gene expression in SaOS-2 cells after exposure to pulsed direct current (30 μA/10 s, square wave) using qRT-PCR at different time points (5, 7,12, and 24 h). MATERIALS AND METHODS: The study investigated the effects of direct current (DC) electrical. stimulation on SaOS-2 cells by exposing experimental. groups to DC (30 μA, 10-s pulses) for 5, 7, 12, and 24 h, while control groups received no stimulation. Stainless steel electrodes were utilized, and both groups were cultured under identical. conditions. Qualitative assessments included cell morphology analysis through phase contrast microscopy, while quantitative evaluations involved MTT assays for cell viability and quantitative reverse transcription PCR (qRT-PCR) for osteoprotegerin (OPG) gene expression. RNA was isolated after stimulation, followed by complementary DNA (cDNA) synthesis for gene analysis. Data were analyzed to assess stimulation-induced cellular and genetic responses. RESULTS: Direct current stimulation induced time-dependent cytotoxicity in SaOS-2 cells, with cell death increasing from approximately 10 % at 5 h to about 52 % at 24 h qRT-PCR reveals significant downregulation of OPG expression, nearly eliminated between 12 and 24 h (p < 0.0001), indicating strong inhibitory effects on both cell viability and gene expression. CONCLUSION: Direct electrical. stimulation downregulated OPG expression in SaOS-2 cells in a time-dependent manner, with a significant decrease observed as early as 5 h. The MTT assay reveals time-dependent cytotoxicity from DC stimulation. Reduced OPG expression suggests potential. enhancement of osteoclastic activity, indicating a possible role of DC stimulation in bone remodelling, which warrants further investigation.