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© A. Alanio, E. Perret
Prolifération de Cryptococcus neoformans dans des macrophages murins.
Publication : Toxicology and applied pharmacology

A cell impedance-based real-time in vitro assay to assess the toxicity of amphotericin B formulations.

Scientific Fields
Diseases
Organisms
Applications
Technique

Published in Toxicology and applied pharmacology - 01 Nov 2017

Menotti J, Alanio A, Sturny-Leclère A, Vitry S, Sauvage F, Barratt G, Bretagne S,

Link to Pubmed [PMID] – 28865757

Link to DOI – S0041-008X(17)30356-310.1016/j.taap.2017.08.017

Toxicol. Appl. Pharmacol. 2017 11; 334(): 18-23

Aerosolized liposomal amphotericin B (L-AmB) has been investigated as prophylaxis against invasive aspergillosis. However, the clinical results are controversial and some trials suggest that toxicity could be a limitation for wider use. Our aim was to assess the dynamics of cell toxicity induced in a human alveolar epithelial cell line (A549) after exposure to L-AmB (50 to 400μg/ml) or amphotericin B deoxycholate (D-AmB; 50 to 200μg/ml) by monitoring real-time A549 cell viability using an impedance-based technology. Results were expressed as cell index values integrating cell adhesion, proliferation, and survival. In parallel, the gene expression of proinflammatory cytokines was quantified at 6 and 24h after drug addition by real-time RT-PCR on cell lysates. No sustained reduction of cell indexes was observed with L-AmB or empty liposomes, even at 400μg/ml. Only the highest concentration tested of L-AmB (400μg/ml) yielded transient significant 6-fold and 4-fold induction of TNF-α and IL-8 mRNAs, respectively. In contrast, D-AmB induced a decrease in cell indexes and only the 50μg/ml concentration of D-AmB was followed by cell recovery, higher concentrations leading to cell death. Significant 4-fold, 7-fold and 3-fold inductions of TNF-α, IL-8 and IL-33 mRNAs were also observed at 6h with 50μg/ml of D-AmB. In conclusion, continuous cell impedance measurement showed no toxicity on overall cellular behavior although a slight proinflammatory cytokine expression is possible after L-AmB challenge. Real-time kinetics of cell impedance is an interesting tool for initial screening of cell toxicity.

https://pubmed.ncbi.nlm.nih.gov/28865757