title: Estimation of bisphenol A-Human toxicity by 3D cell culture arrays, high throughput alternatives to animal tests.
abstract: Bisphenol A (BPA) has been widely used for manufacturing polycarbonate plastics and epoxy resins and has been extensively tested in animals to predict human toxicity. In order to reduce the use of animals for toxicity assessment and provide further accurate information on BPA toxicity in humans, we encapsulated Hep3B human hepatoma cells in alginate and cultured them in three dimensions (3D) on a micropillar chip coupled to a panel of metabolic enzymes on a microwell chip. As a result, we were able to assess the toxicity of BPA under various metabolic enzyme conditions using a high-throughput and micro assay; sample volumes were nearly 2,000 times less than that required for a 96-well plate. We applied a total of 28 different enzymes to each chip, including 10 cytochrome P450s (CYP450s), 10 UDP-glycosyltransferases (UGTs), 3 sulfotransferases (SULTs), alcohol dehydrogenase (ADH), and aldehyde dehydrogenase 2 (ALDH2). Phase I enzyme mixtures, phase II enzyme mixtures, and a combination of phase I and phase II enzymes were also applied to the chip. BPA toxicity was higher in samples containing CYP2E1 than controls, which contained no enzymes (IC50, 184±16μM and 270±25.8μM, respectively, p<0.01). However, BPA-induced toxicity was alleviated in the presence of ADH (IC50, 337±17.9μM), ALDH2 (335±13.9μM), and SULT1E1 (318±17.7μM) (p<0.05). CYP2E1-mediated cytotoxicity was confirmed by quantifying unmetabolized BPA using HPLC/FD. Therefore, we suggest the present micropillar/microwell chip platform as an effective alternative to animal testing for estimating BPA toxicity via human metabolic systems.
authors: Central R & D Institute, Samsung Electro-Mechanics Co. (SEMCO), Suwon, Republic of Korea.
authors: Dong Woo
authors: DW
authors: Lee
authors: Research Center for Cell Fate Control, Department of Toxicology, College of Pharmacy, Sookmyung Women's University, Seoul, Republic of Korea.
authors: Woo-Yeon
authors: WY
authors: Oh
authors: Central R & D Institute, Samsung Electro-Mechanics Co. (SEMCO), Suwon, Republic of Korea.
authors: Sang Hyun
authors: SH
authors: Yi
authors: Central R & D Institute, Samsung Electro-Mechanics Co. (SEMCO), Suwon, Republic of Korea.
authors: Bosung
authors: B
authors: Ku
authors: Department of Chemical & Biomedical Engineering, Cleveland State University, Cleveland, OH, USA.
authors: Moo-Yeal
authors: MY
authors: Lee
authors: Center for Environmental Health Sciences, Department of Biomedical and Pharmaceutical Sciences, University of Montana, Missoula, MT, USA.
authors: Yoon Hee
authors: YH
authors: Cho
authors: Research Center for Cell Fate Control, Department of Toxicology, College of Pharmacy, Sookmyung Women's University, Seoul, Republic of Korea. Electronic address: myang@sm.ac.kr.
authors: Mihi
authors: M
authors: Yang
mesh_terms: D000826:Animal Testing Alternatives; D001559:Benzhydryl Compounds / Q000633:toxicity*; D018929:Cell Culture Techniques / Q000379:methods*; D045744:Cell Line, Tumor; D057166:High-Throughput Screening Assays / Q000295:instrumentation* / Q000379:methods; D006801:Humans; D046208:Microchip Analytical Procedures; D010636:Phenols / Q000633:toxicity*; D018675:Toxicity Tests / Q000295:instrumentation* / Q000379:methods; D000098826:Bisphenol A Compounds
chemical_list: D001559:Benzhydryl Compounds; D010636:Phenols; C006780:bisphenol A; D000098826:Bisphenol A Compounds
keywords: 3D cell microarray; Bisphenol A; Cytotoxicity; Metabolism