Impact of Eu(III) on mammalian cells as a function of its speciation

In the case of the accidental release of long-lived radionuclides, e.g., actinides, into the environment, knowledge of their behavior in bio-systems is necessary to asses and to prevent radiological and chemical induced adverse health effects. This includes knowledge of the bioavailability and chemo-/radiotoxicity of these elements for/onto cells, which are governed to a large extent by their speciation [1,2]. In order to gain a better process understanding, we study the interaction of trivalent actinides/lanthanides with mammalian cells on a cellular level combining biochemical and analytical methods. Results of these studies can contribute to the estimation of low dose effects and the development of new decontamination strategies.
The cellular tolerance of FaDu cells (human squamous cell carcinoma cell line) toward Eu(III) as an analog for trivalent actinides as well as its uptake into the cells has been studied as a function of the Eu(III) concentration and nutrient composition. To differentiate between chemotoxic and radiotoxic effects of Eu(III), 152Eu (β-, ε) was applied as radioactive tracer besides europium with natural isotope composition. The Eu(III) speciation in the cell culture media has been investigated by time-resolved laser-induced fluorescence spectroscopy as well as by solubility studies in combination with ultrafiltration, ultracentrifugation, cation and anion analysis. These results are used to correlate cytotoxicity and uptake of Eu(III) on/into the cells with its chemical speciation in the nutrient. Presently, we are studying the interaction of Eu(III) with NRK-52E cells (rat kidney epithelial-like cells). The results of these studies will be discussed and compared to those obtained with FaDu cells.
From the studies with FaDu cells it was concluded that the Eu(III) cytotoxicity onto these cells depends on the Eu(III) concentration and is influenced by its chemical speciation. This was also reported, for instance, for the toxicity of U(VI) onto rat kidney cells [3]. In the presence of fetal bovine serum (FBS) as nutrient component, Eu(III) is stabilized in solution by complexation with serum proteins as strong complexing agents and shows a low cytotoxicity. In contrast to that, in the absence of FBS, Eu(III) forms hardly soluble species that are dominated by phosphate ligands. In this binding form, Eu(III) exerts a significantly higher cytotoxicity. The presence of an excess of citrate, as strong complexing ligand, influences the Eu(III) speciation and decreases its toxicity in the absence of FBS. Independent of its speciation, Eu(III) seems to be predominantly bound to the cell surface and does not significantly enter the cells. Under the applied experimental conditions, the tolerance of FaDu cells versus Eu(III) appears to be not significantly influenced by the presence of 152Eu, indicating no additional radiotoxic effect.

[1] Ansoborlo, E. et al., Biochimie 88, 1605 (2008).
[2] Bresson, C. et al., J. Anal. At. Spectrom. 26, 593 (2011).
[3] Carrière, M. et al., Chem. Res. Toxicol. 17, 446 (2004).