TUBITAK Destekli Projelerden Çıkan Yayınlar Topluluğu
Published January 1, 2022 | Version v1
Journal article Open

In vitro/in silico prediction of drug induced steatosis in relation to oral doses and blood concentrations by the Nile Red assay

Creators

  • 1. Tech Univ Dortmund IfADo, Leibniz Res Ctr Working Environm & Human Factors, Ardeystr 67, D-44139 Dortmund, Germany
  • 2. TU Dortmund Univ, Dept Stat, Vogelpothsweg 87, D-44227 Dortmund, Germany
  • 3. Hlth Res Inst La Fe, Expt Hepatol Unit, Valencia, Spain
  • 4. Simcyp, Sheffield, S Yorkshire, England
  • 5. Med Univ Vienna, Dept Internal Med 3, Div Gastroenterol & Hepatol, Hans Popper Lab Mol Hepatol, Vienna, Austria
  • 6. Primacyt Cell Culture Technol GmbH, Schwerin, Germany
  • 7. Univ Leipzig, Dept Hepatobiliary Surg & Visceral Transplantat, D-04103 Leipzig, Germany

Description

The accumulation of lipid droplets in hepatocytes is a key feature of drug-induced liver injury (DILI) and can be induced by a subset of hepatotoxic compounds. In the present study, we optimized and evaluated an in vitro technique based on the fluorescent dye Nile Red, further named Nile Red assay to quantify lipid droplets induced by the exposure to chemicals. The Nile Red assay and a cytotoxicity test (CTB assay) were then performed on cells exposed concentration-dependently to 60 different compounds. Of these, 31 were known to induce hepatotox-icity in humans, and 13 were reported to also cause steatosis. In order to compare in vivo relevant blood con-centrations, pharmacokinetic models were established for all compounds to simulate the maximal blood concentrations (Cmax) at therapeutic doses. The results showed that several hepatotoxic compounds induced an increase in lipid droplets at sub-cytotoxic concentrations. To compare how well (1) the cytotoxicity test alone, (2) the Nile Red assay alone, and (3) the combination of the cytotoxicity test and the Nile Red assay (based on the lower EC10 of both assays) allow the differentiation between hepatotoxic and non-hepatotoxic compounds, a previously established performance metric, the Toxicity Separation Index (TSI) was calculated. In addition, the Toxicity Estimation Index (TEI) was calculated to determine how well blood concentrations that cause an increased DILI risk can be estimated for hepatotoxic compounds. Our findings indicate that the combination of both assays improved the TSI and TEI compared to each assay alone. In conclusion, the study demonstrates that inclusion of the Nile Red assay into in vitro test batteries may improve the prediction of DILI compounds.

Files

bib-5f61563f-7410-4830-9773-ee8a24da1467.txt

Files (445 Bytes)

Name Size Download all
md5:232aae357fa95d3b1e06d9384f45da93
445 Bytes Preview Download