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Embryonic cells, when cultured at high density, are referred to as „micromass” cultures and can be used as a tool to predict the teratogenic potential of xenobiotics in screen test. The rate of proliferation and differentiation can easily be measured quantitatively by simple staining methods with subsequent spectrophotometry. However, reproducibility of the results can be influenced by variability of the technique. To adapt this method in our laboratory systematic series of experiments with limb bud cell cultures were undertaken. Inter-relationships between different variables were checked.
The cytototoxic potential of metronidazole, tinidazole, ronidazole, and ornidazole, using human and rat hepatoma cell lines (HepG2 and FaO) in culture was assessed. The cells were treated with drugs for 24, 48 and 72 h at 37 °C in 5% CO₂ at concentrations of 0.1 to 200 µg/mL. Following the treatment period, the cells were assayed by four independent assays: MTT reduction, neutral red uptake (NRU), total protein content (TPC), and LDH leakage. The results suggest that nitroimidazoles are of low cytotoxic potential (EC₅₀ >200µg/mL). The exception was ronidazole, which demonstrated a distinct endpoint sensitivity related to the species. EC₅₀ (µg/mL) in human cells were: in MTT assay - 196±5.5 and 122±9.3 at 24 and 48 h, respectively, and in NRU assay - 150±1.25 at 72 h. Based on minimal toxic concentrations (EC₂₀) for ronidazole, determined by all methods used in HepG2 cells, it could be concluded that their sensitivity was as follows: MTT>NRU>LDH>TPC.
Medycyna Weterynaryjna
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2010
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tom 66
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nr 08
s.525-529,tab.,bibliogr.
This article is a review of recently published papers dealing with the possible participation of environmental hormonally active compounds (natural and synthetic) in the ethiology of reproductive and other health disorders in animals and man. For some "old" synthetic chemicals (DDT, PCBs, dioxins, diethylstilbestrol, bisphenol A, atrazine and tributyltin) there is a link to the impact on wild animals. It has been hypothesized that in humans these compounds may be responsible for the increased incidence of breast and testicular cancer as well as for reproductive disorders. They generally occur at nontoxic concentrations, but exposure to their mixtures seems to be a major medical challenge. The food and drinking water are the main routes of exposure. During last decades much concern is focused on new groups of chemicals such as brominated flame retardants, phthalates, perfluorinated chemicals, parabens and veterinary drugs (anabolics).
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