Elucidate the Role of TGF‐beta in Prevention and Treatment of Gingival Hyperplasia Using HGF‐1 Cells

Abstract

PurposeThe purpose of this study was to evaluate the effects of nifedipine (calcium channel blocker), phenytoin (antiepileptic drug), and losartan (TGF‐beta inhibitor) on cytotoxicity and ATP levels in human gingival fibroblasts (HGF‐1) and evaluate whether losartan attenuates gingival overgrowth caused by nifedipine and phenytoin.MethodsHGF‐1 cells obtained from ATCC were grown to confluency in D‐MEM media supplemented with PenStrep, and bovine serum (FBS 10%) in a CO2 incubator in T‐flasks. For cytotoxicity (LDH leakage in the medium was measured using the CytoTox 96 Non‐radioactive Cytotoxicity Assay Kit) and ATP (Total % ATP was measured using CellTiter‐Glo Luminescent Cell Viability Assay Kit). For ATP and LDH measurements, cells were treated with various concentrations of nifedipine (5, 10, 25, 50 and 100 μM), phenytoin (10, 25, 50, 100 and 200 μM) and losartan (25, 50, 100, 250 and 500 μM) dissolved in DMSO and incubated for 24 h, 72 h and 7 day periods (1×104 cells/well in 96 well plates, 5% CO2, 37 °C and 95% relative humidity).ResultsResults show that none of the drug concentrations had a significant cytotoxic effect on HGF‐1 cells. No significant increase in ATP levels were observed for losartan. At 24h, nifedipine and phenytoin showed significant increase in percent ATP levels when compared to vehicle control. Percent ATP levels for losartan, nifedipine and phenytoin were in range of 101 ± 4 to 113.5 ± 8.2%; 143.1 ± 16.9 to 148.5 ± 20.4% and 135 ± 11.1 to 138.1 ± 16.5% respectively after 24h treatment.ConclusionIn conclusion, our study suggests that nifeidipine and phenytoin causes cell proliferation as evidenced from increased ATP levels. Concentrations identified from these studies will be used to assess effects of losartan on nifedipine and phenytoin induced gingival overgrowth using various markers involved in gingival overgrowth.Support or Funding InformationThis project is supported by College of Pharmacy, Roseman University of Health Sciences, South Jordan, Utah