Combined effects and toxicological interactions of perfluoroalkyl and polyfluoroalkyl substances mixtures in human liver cells (HepG2)

Abstract

The combined effects and toxicological interactions of perfluoroalkyl and polyfluoroalkyl substances (PFAS) mixtures remain largely unknown even though they occur as complex mixtures in the environment. This study investigated the toxicity of individual and combined PFAS to human liver cell line (HepG2). The Combination Index (CI)-isobologram equation method was used to determine the toxicological interactions of PFAS in binary, ternary and multi-component mixtures. The results indicated that the cytotoxicity of individual PFAS to HepG2 cells increased with increasing carbon chain lengths when separated into non-sulfonated and sulfonated groups. The respective cytotoxicity of PFAS is in the order of PFDA > PFNA > PFOA > PFHpA for perfluoroalkyl carboxylic acids and in the order of PFOS > PFHxS for perfluoroalkane sulfonic acids. The toxicological interaction of PFOS and PFOA with other PFAS clearly showed a different pattern of combined toxicity in HepG2 Cells. The binary, ternary, and multi-component combinations of PFOS with PFOA, PFNA, PFDA, PFHxS, and PFHpA displayed synergistic interactions for almost all inhibitory effect levels tested, whereas, either synergistic or antagonistic effect was observed in mixtures with PFOA. Overall, the pattern of interactions of PFAS mixtures is predominated by synergism, especially at low to medium effect levels; the exceptions to this were the antagonistic interactions found in mixture with PFOA, PFHxS, and PFHpA. These cytotoxicity results may have an implication on the health risk assessment of PFAS mixtures.