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Effect of Some Vitamins on Antioxidant/Prooxidant Parameters in Sodium Fluoride (NaF)-Treated Cell Line (hFOB 1.19)
2017
Yuksek, Veysel | Cetin, Sedat | Usta, Ayse | Komuroglu, Ahmet Ufuk | Dede, Semiha
This study was planned to determine the effect of certain vitamin applications on antioxidant and oxidant parameters in the osteoblast cell line exposed to sodium fluoride in vitro and to evaluate the protective role of certain vitamins against possible toxic effects of fluoride. Cells were replicated in vitro conditions with regular passaging 2-3 times weekly. MTF viability test was used to determine IC50 of NaF (5000μM) and proliferative doses of vitamins (Vitamin A: 10μM, Vitamin D: 10μM, Vitamin E: 60μM, Vitamin C: 100μM) for hFOB 1.19 cells. Cells were sown in flasks as so to be 106. The study groups were identified as control, NaF, vitamins and NaF+vitamins. After incubation for 24 hours, cells treated with trypsin were prepared by freeze/thaw method and MTT viability test, TAS, SOD, GSH, CAT, TOS and MDA analyzes were performed on these samples.In the hFOB 1.19 cell line, TAS levels decreased significantly in the NaF group (p≤0.05), but were close to the control group in NaF+vitamin groups with the exception of vitamin C. However, there was no difference between the groups in terms of GSH level and CAT and SOD activities when the control and NaF groups were compared. It was observed that TOS level increased significantly in the NaF group (p<0.05), decreased in the NaF+vitamin groups and were lower in the NaF+vitamin C and E groups than the control group (p <0.05). While OSI was the highest in the NaF group, no significant difference in MDA level was observed compared with the control group.Conclusion: As a result, it was found that NaF administration in the osteoblast cell line increased oxidative stress and decreased following vitamin application. It was found that the effect of NaF administration in the osteoblast cell line on cell viability was consistent with the oxidative stability and that the vitamin application conformably changed cell viability and oxidative balance.
Show more [+] Less [-]Effect of Lycopene Administration on Necrotic Gene Expression in Renal Epithelial Cell Line (NRK-52E) Exposed to Sodium Fluoride
2020
Cetin, Sedat | Yuksek, Veysel | Dede, Semiha | Usta, Ayse | Taspinar, Mehmet
The aim of this study was to determine the effects of lycopene administration as a protective agent against necrotic damage of NaF, a fluoride compound found to have high cytotoxic effects in the renal epithelial cell. Material- Method: The renal epithelial cell was cultured in DMEM high glucose medium, containing 10%FBS, 1%L-Glutamine (2mM) and 1% penicillin/streptomycin. With the MTT viability test, the non-toxic dose of lycopene (1 µM) and the IC50 value of NaF at the 24th hour was determined to be 3200 µM. The study groups were divided into four as control, NaF, lycopene and NaF+lycopene (the combination of NaF and lycopene). After the total mRNA obtained from these groups were converted to cDNA, expression levels of the identified necrotic genes were determined by real-time PCR method.While the Ripk1 gene did not change in the group given lycopene at the 24th hour, it was found that it increased 2.6 times in the group that received only fluoride, while it increased 7 times in the group treated with NaF+lycopene. A significant difference was detected between the groups in terms of gene expression pattern. While the Ripk3 gene increased slightly in the 24th hour applied lycopene group, it was observed that only NaF applied group increased 8 times and NaF+lycopene applied group increased in the 9 times.Based on the results obtained from this study, it was seen that activation of necrotic genes is important in explaining the molecular basis of cell death from NaF, which is applied as fluoride source, in revealing the molecular basis of the necrotic pathway. It was found that the decrease in cell viability due to NaF increased with lycopene, but the use of lycopene with fluoride also increased necrotic gene expression.
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