Research

In vitro effects of nanoparticles on renal cells

Béatrice L'Azou¹ , Joana Jorly¹ , Dinhill On¹ , Elisabeth Sellier² , Frédéric Moisan³ , Jocelyne Fleury-Feith³ , Jean Cambar¹ , Patrick Brochard¹ and Céline Ohayon-Courtès¹

¹  Laboratoire Santé – Travail – Environnement, EA 3672, Université de Bordeaux, Victor Segalen Bordeaux 2,146 rue Léo-Saignat, 33 076 Bordeaux Cedex, France

²  CREMEM Centre de Ressources en Microscopie Electronique et Microanalyse, Université de Bordeaux, Avenue des Facultés, 33 405 Talence, France

³  Laboratoire d'Etude des Particules Inhalées, 11-13 rue Georges Eastman, 75103 Paris, France

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Particle and Fibre Toxicology 2008, 5:22doi:10.1186/1743-8977-5-22

Published:	19 December 2008

Abstract

Background

The ability of nanoparticles to cross the lung-blood barrier suggests that they may translocate to blood and to targets distant from their portal of entry. Nevertheless, nanotoxicity in organs has received little attention. The purpose of this study was to evaluate nanotoxicity in renal cells using in vitro models. Various carbon black (CB) (FW2–13 nm, Printex60-21 nm and LB101-95 nm) and titanium dioxide (TiO₂-15 and TiO₂-50 nm) nanoparticles were characterized on size by electron microscopy. We evaluated theirs effects on glomerular mesangial (IP15) and epithelial proximal tubular (LLC-PK₁) renal cells, using light microscopy, WST-1 assay, immunofluorescence labeling and DCFH-DA for reactive oxygen species (ROS) assay.

Results

Nanoparticles induced a variety of cell responses. On both IP15 and LLC-PK₁ cells, the smallest FW2 NP was found to be the most cytotoxic with classic dose-behavior. For the other NPs tested, different cytotoxic profiles were found, with LLC-PK₁ cells being more sensitive than IP15 cells. Exposure to FW2 NPs, evidenced in our experiments as the most cytotoxic particle type, significantly enhanced production of ROS in both IP15 and LLC-PK₁ cells. Immunofluorescence microscopy using latex beads indicated that depending on their size, the cells internalized particles, which accumulated in the cell cytoplasm. Additionally using transmission electronic microscope micrographs show nanoparticles inside the cells and trapped in vesicles.

Conclusion

The present data constitute the first step towards determining in vitro dose effect of manufactured CB and TiO₂ NPs in renal cells. Cytotoxicological assays using epithelial tubular and glomerular mesangial cell lines rapidly provide information and demonstrated that NP materials exhibit varying degrees of cytotoxicity. It seems clear that in vitro cellular systems will need to be further developed, standardized and validated (relative to in vivo effects) in order to provide useful screening data about the relative toxicity of nanoparticles.