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Title: Microscale 3D Liver Bioreactor for In Vitro Hepatotoxicity Testing under Perfusion Conditions

Abstract

The accurate prediction of hepatotoxicity demands validated human in vitro models that can close the gap between preclinical animal studies and clinical trials. In this study we investigated the response of primary human liver cells to toxic drug exposure in a perfused microscale 3D liver bioreactor. The cellularized bioreactors were treated with 5, 10, or 30 mM acetaminophen (APAP) used as a reference substance. Lactate production significantly decreased upon treatment with 30 mM APAP (p < 0.05) and ammonia release significantly increased in bioreactors treated with 10 or 30 mM APAP (p < 0.0001), indicating APAP-induced dose-dependent toxicity. The release of prostaglandin E2 showed a significant increase at 30 mM APAP (p < 0.05), suggesting an inflammatory reaction towards enhanced cellular stress. The expression of genes involved in drug metabolism, antioxidant reactions, urea synthesis, and apoptosis was differentially influenced by APAP exposure. Histological examinations revealed that primary human liver cells in untreated control bioreactors were reorganized in tissue-like cell aggregates. These aggregates were partly disintegrated upon APAP treatment, lacking expression of hepatocyte-specific proteins and transporters. In conclusion, our results validate the suitability of the microscale 3D liver bioreactor to detect hepatotoxic effects of drugs in vitro under perfusion conditions.

Authors:
 [1];  [1];  [1];  [1];  [1];  [2];  [2];  [3];  [3];  [4];  [1]
  1. Charité – Universitätsmedizin Berlin (Germany). Berlin-Brandenburg Center for Regenerative Therapies (BCRT)
  2. Leipzig Univ. (Germany). Dept. of Hepatobiliary Surgery and Visceral Transplantation
  3. Los Alamos National Lab. (LANL), Los Alamos, NM (United States)
  4. StemCell Systems GmbH, Berlin (Germany)
Publication Date:
Research Org.:
Los Alamos National Lab. (LANL), Los Alamos, NM (United States); Charité – Universitätsmedizin Berlin (Germany); Leipzig Univ. (Germany)
Sponsoring Org.:
USDOE; Defense Threat Reduction Agency (DTRA) (United States); German Federal Ministry of Education and Research (BMBF)
OSTI Identifier:
1479987
Report Number(s):
LA-UR-18-21816
Journal ID: ISSN 2306-5354
Grant/Contract Number:  
AC52-06NA25396; DTRA100271A5196; FKZ 0315741
Resource Type:
Accepted Manuscript
Journal Name:
Bioengineering
Additional Journal Information:
Journal Volume: 5; Journal Issue: 1; Journal ID: ISSN 2306-5354
Publisher:
MDPI
Country of Publication:
United States
Language:
English
Subject:
60 APPLIED LIFE SCIENCES; 42 ENGINEERING; microscale 3D liver bioreactor; in vitro perfusion; primary human liver cells; hepatotoxicity; acetaminophen

Citation Formats

Freyer, Nora, Greuel, Selina, Knöspel, Fanny, Gerstmann, Florian, Storch, Lisa, Damm, Georg, Seehofer, Daniel, Foster Harris, Jennifer, Iyer, Rashi, Schubert, Frank, and Zeilinger, Katrin. Microscale 3D Liver Bioreactor for In Vitro Hepatotoxicity Testing under Perfusion Conditions. United States: N. p., 2018. Web. doi:10.3390/bioengineering5010024.
Freyer, Nora, Greuel, Selina, Knöspel, Fanny, Gerstmann, Florian, Storch, Lisa, Damm, Georg, Seehofer, Daniel, Foster Harris, Jennifer, Iyer, Rashi, Schubert, Frank, & Zeilinger, Katrin. Microscale 3D Liver Bioreactor for In Vitro Hepatotoxicity Testing under Perfusion Conditions. United States. doi:10.3390/bioengineering5010024.
Freyer, Nora, Greuel, Selina, Knöspel, Fanny, Gerstmann, Florian, Storch, Lisa, Damm, Georg, Seehofer, Daniel, Foster Harris, Jennifer, Iyer, Rashi, Schubert, Frank, and Zeilinger, Katrin. Thu . "Microscale 3D Liver Bioreactor for In Vitro Hepatotoxicity Testing under Perfusion Conditions". United States. doi:10.3390/bioengineering5010024. https://www.osti.gov/servlets/purl/1479987.
@article{osti_1479987,
title = {Microscale 3D Liver Bioreactor for In Vitro Hepatotoxicity Testing under Perfusion Conditions},
author = {Freyer, Nora and Greuel, Selina and Knöspel, Fanny and Gerstmann, Florian and Storch, Lisa and Damm, Georg and Seehofer, Daniel and Foster Harris, Jennifer and Iyer, Rashi and Schubert, Frank and Zeilinger, Katrin},
abstractNote = {The accurate prediction of hepatotoxicity demands validated human in vitro models that can close the gap between preclinical animal studies and clinical trials. In this study we investigated the response of primary human liver cells to toxic drug exposure in a perfused microscale 3D liver bioreactor. The cellularized bioreactors were treated with 5, 10, or 30 mM acetaminophen (APAP) used as a reference substance. Lactate production significantly decreased upon treatment with 30 mM APAP (p < 0.05) and ammonia release significantly increased in bioreactors treated with 10 or 30 mM APAP (p < 0.0001), indicating APAP-induced dose-dependent toxicity. The release of prostaglandin E2 showed a significant increase at 30 mM APAP (p < 0.05), suggesting an inflammatory reaction towards enhanced cellular stress. The expression of genes involved in drug metabolism, antioxidant reactions, urea synthesis, and apoptosis was differentially influenced by APAP exposure. Histological examinations revealed that primary human liver cells in untreated control bioreactors were reorganized in tissue-like cell aggregates. These aggregates were partly disintegrated upon APAP treatment, lacking expression of hepatocyte-specific proteins and transporters. In conclusion, our results validate the suitability of the microscale 3D liver bioreactor to detect hepatotoxic effects of drugs in vitro under perfusion conditions.},
doi = {10.3390/bioengineering5010024},
journal = {Bioengineering},
number = 1,
volume = 5,
place = {United States},
year = {2018},
month = {3}
}

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