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Influence of uranium(VI) speciation for the evaluation of in vitro uranium cytotoxicity on LLC-PK1 cells

Institut de Protection et de SûretéNucléaire, Département de Protection de la santé de l'Homme et de Dosimétrie, Service de Dosimétrie, Laboratoire d'Etudes Appliquées de Radiotoxicologie, IPSN, BP 38, F 26701 Pierrelatte Cedex, France

M P Barrouillet

Groupe d'Etude de Physiologie et Physiopathologie Rénales. Faculté de Pharmacie- 146, Rue Léo Saignat- 33076 Bordeaux Cedex, France

M H Hengé-Napoli

Institut de Protection et de SûretéNucléaire, Département de Protection de la santé de l'Homme et de Dosimétrie, Service de Dosimétrie, Laboratoire d'Etudes Appliquées de Radiotoxicologie, IPSN, BP 38, F 26701 Pierrelatte Cedex, France

E Ansoborlo

Institut de Protection et de SûretéNucléaire, Département de Protection de la santé de l'Homme et de Dosimétrie, Service de Dosimétrie, Laboratoire d'Etudes Appliquées de Radiotoxicologie, IPSN, BP 38, F 26701 Pierrelatte Cedex, France

M Fournier

Université des Sciences et Technologies de Lille. Laboratoire de catalyse homogène et hétérogène, URA CNRS 402.59655 Villeneuve d'Ascq Cedex, France

J Cambar

Groupe d'Etude de Physiologie et Physiopathologie Rénales. Faculté de Pharmacie- 146, Rue Léo Saignat- 33076 Bordeaux Cedex, France

Very few data are available concerning the in vitro toxicity of uranium. In this work, we have determined the experimental chemical conditions permitting the observation of uranium(VI) cytotoxicity on LLC-PK₁ cells. Uranium solutions made either by dissolving uranyl acetate or nitrate crystals, or by complexing uranium with bicarbonate, phosphate or citrate ligands, were prepared and tested. Experiments demonstrated that only uranium solutions containing citrate and bicarbonate ligands concentrations tenfold higher than the metal, were soluble in the cell culture medium. Cytotoxicity studies of all these uranium compounds were performed on LLC-PK₁ cells and compared using LDH release, neutral red uptake and MTT assays. Dose dependent cytotoxicity curves were only obtained with uranium-bicarbonate medium. This study has revealed a toxicity of uranium-bicarbonate complexes for 24 h expositions and for concentrations ranging from 7x10^-4- 10^-3 M, under these conditions, the CI₅₀ (cytotoxicity index) was evaluated between 8.5 and 9x10^-4 M. In contrast, we noticed a lack of cytotoxicity response for uranium(VI)-citrate complexes. Electron transmission microscopy studies revealed, when LLC-PK₁ cells were exposed to the uranium-bicarbonate system, that uranium penetrated and precipitated within the cytoplasmic compartment. Morphological studies conducted with citrate complexes did not show any cellular intake of uranium.

Key Words: cell culture • cytotoxicity • precipitation • speciation • uranium

Human & Experimental Toxicology, Vol. 18, No. 3, 180-187 (1999)
DOI: 10.1177/096032719901800308


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