DETERMINATION OF DRUG TOXICITY OF ACETAMINOPHEN USING 3D SCAFFOLD CULTURES OF IMMORTAL HUMAN HEPATOCARCINOMA CELL LINES

Acetaminophen (APAP), well-known as paracetamol, is a safe analgesic and antipyretic agent at a therapeutic dose. However, overdoses of APAP can induce hepa­totoxicity, which in turn causes severe liver injury. Various hepatic models mimicking liver architecture have been investigated to examine the potential hepatotoxic effects of chemicals and drugs, but there is always a demand for in-vitro high-throughput hepatic platforms for measuring the hepatotoxicity. This study aims to provide a simple, easy to fabricate and work, a micro-space three-dimensional (3D) scaffold culture system as an in-vitro model for APAP induced hepatotoxicity studies on hepatocarcinoma cell lines (HCC – HepG2 and Huh-7). A silicon mould based template was used to cast out polydimethylsiloxane (PDMS) hexagonal scaffolds. Cytotoxicity was performed by MTT assay. Fluorescence microscopy is used to know the differences in the morphology of the cells grown on scaffolds. mRNA levels of cytochrome P450 2E1 (CYP2E1) expressions were demonstrated through qRT-PCR technique. This study examined the characteristics and usefulness of HepG2 and Huh-7 cell lines grown on the PDMS scaffold system, a three-dimensional culture method as an in vitro human model for APAP-induced hepatotoxicity studies. Scaffold cultured HepG2 and Huh-7 cells showed higher expression of mRNA levels of CYP2E1, more susceptibility for APAP. We have verified that in-house developed PDMS scaffold culture method for APAP toxicity, by measuring cytotoxicity studies, measuring DNA proliferation, and by showing the expression of CYP2E1 in our system.