An in vitro model for long-term hepatotoxicity testing utilizing rat hepatocytes entrapped in micro-hollow fiber reactor

Abstract A long-term hepatocyte model in vitro is preferable for chronic hepatotoxicity research because hepatocytes in this model of culture can preserve liver-specific functions for long period. Micro-hollow fiber reactors (MHFR), composed of polysulphone (PS) hollow fibers with a molecular weight cut-off 100 kDa, were applied to test the hepatotoxicity of acetaminophen, isoniazid and rifampicin, respectively. Monolayer culture was used as a control model for hepatocyte culture. It was found that hepatocytes within MHFR were more sensitive to toxicity of acetaminophen (0.38–1.51 g/L) than those in monolayer cultures. Furthermore, significant hepatotoxicity of isoniazid (15 mg/L) and rifampicin (10 mg/L) were detected in hepatocytes cultured in MHFR but not detected in hepatocyte monolayer, which could be due to well-preserved drug metabolizing enzymes in MHFR. These results indicate that the MHFR may be an effective model for long-term hepatotoxicity research in vitro.

[1]  R. Tompkins,et al.  Long‐Term in Vitro Function of Adult Hepatocytes in a Collagen Sandwich Configuration , 1991, Biotechnology progress.

[2]  C. Westmoreland,et al.  The differential cytotoxicity of methotrexate in rat hepatocyte monolayer and spheroid cultures. , 2000, Toxicology in vitro : an international journal published in association with BIBRA.

[3]  F. Oesch,et al.  Hepatocytes cultured in alginate microspheres: an optimized technique to study enzyme induction. , 2005, Toxicology.

[4]  D. Girling.  The hepatic toxicity of antituberculosis regimens containing isoniazid, rifampicin and pyrazinamide. , 1977, Tubercle.

[5]  Jinsheng Xu,et al.  Energy metabolism and biotransformation as endpoints to pre-screen hepatotoxicity using a liver spheroid model. , 2006, Toxicology and applied pharmacology.

[6]  M. Toner,et al.  Effect of extracellular matrix topology on cell structure, function, and physiological responsiveness: hepatocytes cultured in a sandwich configuration , 1996, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[7]  S. Tannenbaum,et al.  Characterization of chemically induced hepatotoxicity in collagen sandwiches of rat hepatocytes. , 2005, Toxicological sciences : an official journal of the Society of Toxicology.

[8]  C. Viau,et al.  Cytotoxicity of tacrine and velnacrine metabolites in cultured rat, dog and human hepatocytes. , 1993, Drug and chemical toxicology.

[9]  S. Karthikeyan.  Isoniazid and Rifampicin Treatment on Phospholipids and Their Subfractions in Liver Tissue of Rabbits , 2005, Drug and chemical toxicology.

[10]  T. Vanhaecke,et al.  Collagen-Gel Cultures of Rat Hepatocytes: , 1998 .

[11]  M. Tsai,et al.  Using MTT viability assay to test the cytotoxicity of antibiotics and steroid to cultured porcine corneal endothelial cells. , 1996, Journal of ocular pharmacology and therapeutics : the official journal of the Association for Ocular Pharmacology and Therapeutics.

[12]  M. Rissel,et al.  Toxic Effects of Tacrine on Primary Hepatocytes and Liver Epithelial Cells in Culture , 1998, Cell Biology and Toxicology.

[13]  J. Barbet,et al.  Protective effect of stiripentol on acetaminophen-induced hepatotoxicity in rat. , 2001, Toxicology and applied pharmacology.

[14]  Hui Li,et al.  Evaluation of diffusion in gel entrapment cell culture within hollow fibers. , 2005, World journal of gastroenterology.

[15]  T.M Walker,et al.  Cytochrome P450 activity in control and induced long-term cultures of rat hepatocyte spheroids. , 2001, Toxicology in vitro : an international journal published in association with BIBRA.

[16]  R. Panchagnula,et al.  Comparative bioavailability of rifampicin, isoniazid and pyrazinamide from a four drug fixed dose combination with separate formulations at the same dose levels. , 2004, International journal of pharmaceutics.

[17]  S. Nyberg,et al.  Extracorporeal Application of a Gel-Entrapment, Bioartificial Liver: Demonstration of Drug Metabolism and Other Biochemical Functions , 1993, Cell transplantation.

[18]  W. Yew.  Clinically Significant Interactions with Drugs Used in the Treatment of Tuberculosis , 2002, Drug safety.

[19]  T. Harauchi,et al.  Effect of P-450 inducers on glutathione (GSH) depletion by bromobenzene in primary cultures of dog hepatocytes. , 1994, Biological & pharmaceutical bulletin.

[20]  G. Hamilton,et al.  The use of liver spheroids as an in vitro model for studying induction of the stress response as a marker of chemical toxicity. , 2000, Toxicology in vitro : an international journal published in association with BIBRA.

[21]  S. Rana,et al.  Isoniazid – and rifampicin–induced oxidative hepatic injury – protection by N–acetylcysteine , 2000, Human & experimental toxicology.

[22]  Wei-Shou Hu,et al.  Characterization of a hollow fiber bioartificial liver device. , 2005, Artificial organs.

[23]  M. Czaja,et al.  Cytochrome P450 2E1 expression induces hepatocyte resistance to cell death from oxidative stress. , 2002, Antioxidants & redox signaling.

[24]  Jos H Beijnen,et al.  An update on in vitro test methods in human hepatic drug biotransformation research: pros and cons. , 2003, Toxicology and applied pharmacology.

[25]  Ai-Ming Yu,et al.  THE CYP2E1-HUMANIZED TRANSGENIC MOUSE: ROLE OF CYP2E1 IN ACETAMINOPHEN HEPATOTOXICITY , 2005, Drug Metabolism and Disposition.

[26]  T. Maguire,et al.  Progressive maturation resistance to microcystin-LR cytotoxicity in two different hepatospheroidal models , 1999, Cell Biology and Toxicology.

[27]  F. Oesch,et al.  New Hepatocyte In Vitro Systems for Drug Metabolism: Metabolic Capacity and Recommendations for Application in Basic Research and Drug Development, Standard Operation Procedures , 2003, Drug metabolism reviews.

[28]  H. Jaeschke,et al.  Acetaminophen-induced oxidant stress and cell injury in cultured mouse hepatocytes: protection by N-acetyl cysteine. , 2004, Toxicological sciences : an official journal of the Society of Toxicology.

[29]  S. Nyberg,et al.  Characterization of the three-compartment gel-entrapment porcine hepatocyte bioartificial liver , 1997, Cell Biology and Toxicology.

[30]  Christopher S. Chen,et al.  Engineering cellular microenvironments to improve cell-based drug testing. , 2002, Drug discovery today.

[31]  F. Alvarez,et al.  Long-term culture of adult rat hepatocyte spheroids. , 1992, Experimental cell research.