The cellular and molecular pathways of fluoride toxicity in osteoblasts are not very well understood. Therefore, the objective of the present study was to evaluate the effects of sodium fluoride (NaF) on caprine osteoblasts cultured in vitro. Caprine osteoblasts at 2.0 x 10(-4) cells/ml were incubated in vitro with NaF at 0, 10(-8), 10(-7), 10(-6), 10(-5), 10(-4), 5.0 x 10(-4), and 10(-3) M, and then proliferation, differentiation, apoptosis, calcification, and alkaline phosphatase activity were examined. Also, the effect of NaF on osteoblastic cell viability and the molecular events leading to apoptosis were determined. Electron microscopy revealed cytoplasmic and nuclear alterations in the ultrastructure of osteoblasts exposed to various NaF concentrations. A cell-based quantitative evaluation of the MTT assay showed that NaF at concentrations of 10(-8) to 10(-5) M promoted cell proliferation, whereas at 10(-4) to 10(-3) M it suppressed cell proliferation and induced apoptosis. Alkaline phosphatase (ALP) activity and mineralization ability increased in cells treated at 10(-8) to 10(-5) M with sodium versus the controls, but decreased at 5.0 x 10(-4) to 10(-3) M dosage. The highest incidence of early apoptotic cells and late apoptotic cells was reached (3.33% and 2.92%, respectively) under NaF concentration of 10(-4) M. In conclusion, results of this study indicated that NaF modulates osteoblast proliferation and differentiation in a dose-dependent manner and modified osteoblast metabolism bidirectionally, suggesting NaF may play a significant role in osteoblast physiology.
The antioxidant enzymes, lipid peroxidation and free radicals assessment were made of the effects of selenium, copper and magnesium on bovine endemic fluorosis under high fluoride, low selenium and low copper productive conditions. Thirty-two beef cattle were selected from high fluoride area, and randomly divided into four groups with eight cattle each as follows: (1) high fluoride control group (HFC); (2) supplemented group with 0.25 mg/kg selenium (HFSe); (3) supplemented group with 15 mg/kg copper (HFCu) and (4) supplemented group with 0.25 mg/kg selenium+15 mg/kg copper+1 mg/kg magnesium (HFSeCuMg) per day for 83 days. Moreover, eight beef cattle were selected from non-high fluoride area as normal control group. Blood samples were collected from cattle on 0 d, 30 d and 83 d respectively, to analyze the enzyme activities and concentration of GSH-px, CAT, SOD, MDA and free radicals. The results showed that the contents of free radicals and MDA in HFC group were significantly higher, and the whole blood GSH-px, CAT, erythrocyte SOD activities were lower than the normal control group. Free radicals, metabolic imbalance and antioxidant disorder therefore, play an important role in fluorosis. However, GSH-px, CAT and SOD activities in HFSe group and HFSeCuMg group at 30 d and 83 d were markedly higher than the same groups at the 0 d and the HFC group at the same time. Likewise, there was a corresponding reduction in the contents of free radicals and MDA. These findings indicated that supplementation with selenium, copper and magnesium elevated high fluoride bovine antioxidant enzymes, and decreased MDA and free radicals contents. But, the activities of supplementation selenium group did not increase until day 83. These results demonstrated that fluorosis was associated with lower serum Se and Cu levels than in the control, and it was therefore concluded that fluorosis is associated with decreased serum levels of these minerals. Long-term high fluoride intake under productive condition enhances oxidative stress in the blood, thereby disturbing the antioxidant defense of cattle. Increased oxidative stress could be one of the mediating factors in the pathogenesis of toxic manifestations of fluoride. It is benefical for high fluoride cattle supplemented with proper selenium, copper and magnesium to increase fluoride excretion and obtain the protective impact of the activity of oxidative enzymes, and to decrease lipid peroxidation and free radicals contents.
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