Comparative in vitro toxicity assessment of perfluorinated carboxylic acids

• Published in: Journal of applied toxicology 2017-06, Vol.37 (6), p.699-708
• Main Authors: Mahapatra, Cecon T., Damayanti, Nur P., Guffey, Samuel C., Serafin, Jennifer S., Irudayaraj, Joseph, Sepúlveda, Maria S.
• Format: Article
• Language: English
• Subjects: Animals; Biological Transport; Biomarkers - metabolism; Caproates - toxicity; Caprylates - toxicity; Cell Line; Danio rerio; Dose-Response Relationship, Drug; Environmental Pollutants - toxicity; Fluorescence Lifetime Imaging Microscopy; Fluorocarbons - toxicity; Gene Expression - drug effects; Lipid Metabolism - drug effects; Lipid Metabolism - genetics; Liver; Liver - drug effects; Liver - metabolism; Liver - pathology; Microscopy, Fluorescence; Perfluoroalkyl substances; Perfluorobutanoic acid; Perfluorohexanoic acid; Structure-Activity Relationship; Zebrafish
• ISSN: 0260-437X; 1099-1263
• DOI: 10.1002/jat.3418

Perfluoroalkyl and polyfluoroalkyl substances (PFASs) are synthetic fluorinated compounds that are highly bioaccumulative and persistent organic pollutants. Perfluorooctanoic acid (PFOA), an eight‐carbon chain perfluorinated carboxylic acid, was used heavily for the production of fluoropolymers, but concerns have led to its replacement by shorter carbon chain homologues such as perfluorohexanoic acid (PFHxA) and perfluorobutanoic acid (PFBA). However, limited toxicity data exist for these substitutes. We evaluated the toxicity of PFOA, PFHxA and PFBA on a zebrafish liver cell line and investigated the effects of exposure on cell metabolism. Gross toxicity after 96 h of exposure was highest for PFOA and PFO–, while PFHxA and PFBA exhibited lower toxicity. Although the structural similarity of these compounds to fatty acids suggests the possibility of interference with the transport and metabolism of lipids, we could not detect any differential expression of peroxisome proliferator‐activated receptor (ppar‐α, ‐β and ‐γ), fabp3 and crot genes after 96 h exposure to up to 10 ppm of the test compounds. However, we observed localized lipid droplet accumulation only in PFBA‐exposed cells. To study the effects of these compounds on cell metabolism, we conducted fluorescence lifetime imaging microscopy using naturally fluorescent biomarkers, NADH and FAD. The fluorescence lifetimes of NADH and FAD and the bound/free ratio of each of these coenzymes decreased in a dose‐ and carbon length‐dependent manner, suggesting disruption of cell metabolism. In sum, our study revealed that PFASs with shorter carbon chains are less toxic than PFOA, and that exposure to sublethal dosage of PFOA, PFHxA or PFBA affects cell metabolism. Copyright © 2016 John Wiley & Sons, Ltd. Perfluorooctanoic acid is a toxic and persistent pollutant, but little is known about the toxicity of shorter chain alternatives. We found that the shorter chain alternatives perfluorohexanoic acid and perfluorobutanoic acid caused less gross toxicity to zebrafish liver cells as compared to perfluorooctanoic acid, but all three compounds affected the fluorescence lifetime of free and bound forms of NADH and FAD, indicating a disruption of cell metabolism.