Measured and modeled toxicokinetics in cultured fish cells and application to in vitro-in vivo toxicity extrapolation.

Julita Stadnicka-Michalak,Roman Ashauer,Kristin Schirmer,Katrin Tanneberger,Maria A Deli

doi:10.1371/journal.pone.0092303

Abstract

Effect concentrations in the toxicity assessment of chemicals with fish and fish cells are generally based on external exposure concentrations. External concentrations as dose metrics, may, however, hamper interpretation and extrapolation of toxicological effects because it is the internal concentration that gives rise to the biological effective dose. Thus, we need to understand the relationship between the external and internal concentrations of chemicals. The objectives of this study were to: (i) elucidate the time-course of the concentration of chemicals with a wide range of physicochemical properties in the compartments of an in vitro test system, (ii) derive a predictive model for toxicokinetics in the in vitro test system, (iii) test the hypothesis that internal effect concentrations in fish (in vivo) and fish cell lines (in vitro) correlate, and (iv) develop a quantitative in vitro to in vivo toxicity extrapolation method for fish acute toxicity. To achieve these goals, time-dependent amounts of organic chemicals were measured in medium, cells (RTgill-W1) and the plastic of exposure wells. Then, the relation between uptake, elimination rate constants, and log KOW was investigated for cells in order to develop a toxicokinetic model. This model was used to predict internal effect concentrations in cells, which were compared with internal effect concentrations in fish gills predicted by a Physiologically Based Toxicokinetic model. Our model could predict concentrations of non-volatile organic chemicals with log KOW between 0.5 and 7 in cells. The correlation of the log ratio of internal effect concentrations in fish gills and the fish gill cell line with the log KOW was significant (r>0.85, p = 0.0008, F-test). This ratio can be predicted from the log KOW of the chemical (77% of variance explained), comprising a promising model to predict lethal effects on fish based on in vitro data.

Highlights

Environmental regulations require comprehensive testing and risk assessment before a chemical can be approved for use
The relation between uptake and elimination rate constants and log KOW was investigated in order to develop a multi-compartment TK model describing the chemical distribution in the exposure well
Based on previous in vitro toxicity studies [6,23], the empirically obtained multi-compartment TK model was used to predict internal effect concentrations in cells (IEC50), which were compared with internal effect concentrations in fish gills (ILC50) predicted by a Physiologically Based Toxicokinetic (PBTK) model [24]

Summary

Introduction

Environmental regulations require comprehensive testing and risk assessment before a chemical can be approved for use. Tanneberger et al [6] highlighted that, because gill epithelia are the primary uptake site of water-born contaminants into fish, they could be a primary target for many toxicants in exposure scenarios where vital epithelial cell functions are destroyed, resulting in a toxic effect on the whole organism. Along these lines, Li et al [7] noticed that in fish, gill tissue can be more sensitive to some chemicals than liver and muscle tissues. For these reasons, understanding the toxicokinetics in gill cells and the resulting improvement of in vitro – in vivo toxicity extrapolations is very important

Objectives

Methods

Results