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Review
. 2021 May 10;22(9):5038.
doi: 10.3390/ijms22095038.

In Vitro Liver Toxicity Testing of Chemicals: A Pragmatic Approach

Andrés Tabernilla  1 , Bruna Dos Santos Rodrigues  1 , Alanah Pieters  1 , Anne Caufriez  1 , Kaat Leroy  1 , Raf Van Campenhout  1 , Axelle Cooreman  1 , Ana Rita Gomes  1 , Emma Arnesdotter  1 , Eva Gijbels  1 , Mathieu Vinken  1
Affiliations
Review

In Vitro Liver Toxicity Testing of Chemicals: A Pragmatic Approach

Andrés Tabernilla et al. Int J Mol Sci. .

Abstract

The liver is among the most frequently targeted organs by noxious chemicals of diverse nature. Liver toxicity testing using laboratory animals not only raises serious ethical questions, but is also rather poorly predictive of human safety towards chemicals. Increasing attention is, therefore, being paid to the development of non-animal and human-based testing schemes, which rely to a great extent on in vitro methodology. The present paper proposes a rationalized tiered in vitro testing strategy to detect liver toxicity triggered by chemicals, in which the first tier is focused on assessing general cytotoxicity, while the second tier is aimed at identifying liver-specific toxicity as such. A state-of-the-art overview is provided of the most commonly used in vitro assays that can be used in both tiers. Advantages and disadvantages of each assay as well as overall practical considerations are discussed.

Keywords: cytotoxicity; in vitro; liver; liver-specific toxicity; mechanisms.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Scheme representing the structure of the liver, highlighting the cellular architecture of its functional unit, the acinus.
Figure 2
Figure 2
Scheme representing the mechanisms of general cytotoxicity and corresponding in vitro assays. ATP: adenosine triphosphate; Calcein-AM: calcein-acetoxymethyl; DCFH2-DA: 2′,7′-dichlorodihydrofluorescein diacetate; DHE: dihydroethidium; LDH: lactate dehydrogenase; MDA: malondialdehyde; PI: propidium iodide; TBARS: thiobarbituric acid reactive substance; TUNEL: terminal deoxynUcleotidyl transferase dUTP Nick End Labeling.
Figure 3
Figure 3
Schematic representation of the 3 main types of liver-specific toxicity and corresponding in vitro assays. BA: bile acid; BODIPY 493/503: 4,4-difluoro-1,3,5,7,8-pentamethyl-4-bora-3a,4a-diaza-s-indacene 495/503; CCK8: cholecystokinin-octapeptide; CDFDA: 5(6)-carboxy-2′,7′-dichlorofluorescein diacetate; CLF: cholyl-lysyl-fluorescein; FA: fatty acid; MMPs: matrix metalloproteinases; HSC: hepatic stellate cell; Tauro-nor-THCA-24-DBD: tauro-nor-N-(24-[7-(4-N,N-dimethylaminosulfonyl-2,1,3-benzoxadiazole)]-amino-3α,7α,12α-trihydroxy-27-nor-5β-cholestan-26-oyl)-2′-aminoethanesulfonate; TCA: taurocholate; TIMPs: tissue inhibitor of metalloproteinases; α-SMA: alpha-smooth muscle actin.
Figure 4
Figure 4
Diagram summarizing critical parameters for in vitro liver toxicity testing.
See this image and copyright information in PMC

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