Real-time monitoring of cadmium toxicity in rabbit kidney cells

Abstract

The aim of this study was to investigate the toxic effect of the metal salt cadmium chloride dihydrate on the rabbit kidney cell line using the xCELLigence system or real-time cell analyser (RTCA), and to compare this relatively new method with standard biological cytotoxicity assays. This system provides real-time monitoring of cell behaviour and proliferative activity during the whole time of experiment. Moreover, after 24 h exposure of cells to cadmium, colorimetric 3-[4,5-dimethylthiazol-2-yl]-2,5-difenyl tetrazolium bromide (MTT) test was used to measure the metabolic activity and cytotoxicity was determined by measurement of lactate dehydrogenase (LDH) leaked from damaged cells. We found that renal cells exposed to lower concentrations (5-10 mg·l -1 ) of cadmium tend to grow similarly to control cells, however, cell index was significantly different (P < 0.05) after 24 h. With increasing concentration of cadmium (15-50 mg·l -1 ) significantly lower proliferative (P < 0.05) and metabolic activity (P < 0.05) of cells was observed and cytotoxicity increased simultaneously (P < 0.001). In addition, we found that the real-time monitoring of the cell response was significantly correlated with commonly used biological methods for toxicity measurement, for MTT assay R 2 was 0.9448 (P < 0.01) and for LDH assay R 2 was 0.9466 (P < 0.01), respectively. The present study is the first report when combination of RTCA, MTT assay and LDH test was used for cadmium nephrotoxicity assessment. In all these methods, the toxic effect of cadmium on rabbit kidney cells increased in a concentration-dependent manner. xCELLigence system, nephrotoxicity, cytotoxicity, lactate dehydrogenaseIncreased industrial activities brought gradual redistribution of a number of toxic elements from the terrestrial crust into the environment and thus increased the potential exposure of humans and animals. With rising environmental pollution the interest arises in the consequences of the action of xenobiotics, including hazardous chemical elements, on live organisms (Nováková et al. 2007;Čelechovská et al. 2008). A series of trials were designed to evaluate the responses of tissues and organs of animal species of economic importance, such as the rabbit, to trace elements (cadmium, lead, mercury) at concentrations that could potentially exist in the feeds on a farm. The daily rations included carrots, potatoes, or beetroot. Cadmium belongs to heavy metals widely distributed in the environment, obtained as a by-product of refining. Its compounds are used in the electroplating of metals, alkaline batteries or in compounds with other metals. Relatively high quantities of cadmium are present in phosphate fertilisers (from some locations), which increase the concentration of cadmium in soil and plants. Cadmium is also highly toxic to organisms living in the aqueous environment (Koréneková et al. 2002). Because of its ability to reabsorb and accumulate divalent metals, kidney is the first target organ of heavy metal toxicity. The extent of renal da...

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