Cytotoxicity of pharmaceuticals and their mixtures toward scaffold-free 3D spheroid cultures of rainbow trout (Oncorhynchus mykiss) hepatocytes

被引:2
作者
Jarvinen, Paivi [1 ]
Kakko, Maija [1 ]
Sikanen, Tiina [1 ,2 ]
机构
[1] Univ Helsinki, Fac Pharm, Drug Res Program, POB Viikinkaari 5E 56, FI-00014 Helsinki, Finland
[2] Univ Helsinki, Helsinki Inst Sustainabil Sci, POB Yliopistonkatu 3 4, FI-00014 Helsinki, Finland
基金
芬兰科学院;
关键词
Rainbow trout; Hepatocytes; 3D cultures; Spheroids; Hepatotoxicity; Mixture toxicity; Pharmaceuticals; XENOBIOTIC METABOLISM; FISH; TOXICITY; BIOACCUMULATION; KETOCONAZOLE; RISK; EXPRESSION; CYP1A; CYP3A;
D O I
10.1016/j.ejps.2024.106817
中图分类号
R9 [药学];
学科分类号
1007 ;
摘要
Pharmaceutical residues are widely detected in surface waters all around the world, causing a range of adverse effects on environmental species, such as fish. Besides population level effects (mortality, reproduction), pharmaceutical residues can bioaccumulate in fish tissues resulting in organ-specific toxicities. In this study, we developed in vitro 3D culture models for rainbow trout (Oncorhynchus mykiss) liver cell line (RTH-149) and cryopreserved, primary rainbow trout hepatocytes (RTHEP), and compared their spheroid formation and susceptibility to toxic impacts of pharmaceuticals. The rapidly proliferating, immortalized RTH-149 cells were shown to form compact spheroids with uniform morphology in just three days, thus enabling higher throughput toxicity screening compared with the primary cells that required acclimation times of about one week. In addition, we screened the cytotoxicity of a total of fourteen clinically used human pharmaceuticals toward the 3D cultures of both RTH-149 cells (metabolically inactive) and primary RTHEP cells (metabolically active), to evaluate the impacts of the pharmaceuticals' own metabolism on their hepatotoxicity in rainbow trout in vitro. Among the test substances, the azole antifungals (clotrimazole and ketoconazole) were identified as the most cytotoxic, with hepatic metabolism indicatively amplifying their toxicity, followed by fluoxetine, levomepromazine, and sertraline, which were slightly less toxic toward the metabolically active primary cells than RTH-149 spheroids. Besides individual pharmaceuticals, the 3D cultures were challenged with mixtures of the eight most toxic substances, to evaluate if their combined mixture toxicities can be predicted based on individual substances' half-maximal effect (EC50) concentrations. As a result, the classical concentration addition approach was concluded sufficiently accurate for preliminarily informing on the approximate effect concentrations of pharmaceutical mixtures on a cellular level. However, direct read-across from human data was proven challenging and inexplicit for prediction of hepatotoxicity in fish in vitro.
引用
收藏
页数:12
相关论文
共 60 条
[31]   3D human liver spheroids for translational pharmacology and toxicology [J].
Ingelman-Sundberg, Magnus ;
Lauschke, Volker M. .
BASIC & CLINICAL PHARMACOLOGY & TOXICOLOGY, 2022, 130 :5-15
[32]   In vitro biotransformation of pharmaceuticals and pesticides by trout liver S9 in the presence and absence of carbamazepine [J].
Jeon, Junho ;
Hollender, Juliane .
ECOTOXICOLOGY AND ENVIRONMENTAL SAFETY, 2019, 183
[33]  
Jeong Y, 2016, J ENV ANAL TOXICOL, V6, P1, DOI [10.4172/2161-0525.1000404, DOI 10.4172/2161-0525.1000404]
[34]   Predicted exposures to steroid estrogens in UK rivers correlate with widespread sexual disruption in wild fish populations [J].
Jobling, Susan ;
Williams, Richard ;
Johnson, Andrew ;
Taylor, Ayesha ;
Gross-Sorokin, Melanie ;
Nolan, Monique ;
Tyler, Charles R. ;
van Aerle, Ronny ;
Santos, Eduarda ;
Brighty, Geoff .
ENVIRONMENTAL HEALTH PERSPECTIVES, 2006, 114 :32-39
[35]   Development of three-dimensional (3D) spheroid cultures of the continuous rainbow trout liver cell line RTL-W1 [J].
Lammel, Tobias ;
Tsoukatou, Georgia ;
Jellinek, Johanna ;
Sturve, Joachim .
ECOTOXICOLOGY AND ENVIRONMENTAL SAFETY, 2019, 167 :250-258
[36]   Big Question to Developing Solutions: A Decade of Progress in the Development of Aquatic New Approach Methodologies from 2012 to 2022 [J].
Langan, Laura M. ;
Paparella, Martin ;
Burden, Natalie ;
Constantine, Lisa ;
Margiotta-Casaluci, Luigi ;
Miller, Thomas H. ;
Moe, S. Jannicke ;
Owen, Stewart F. ;
Schaffert, Alexandra ;
Sikanen, Tiina .
ENVIRONMENTAL TOXICOLOGY AND CHEMISTRY, 2024, 43 (03) :559-574
[37]   Spheroid Size Does not Impact Metabolism of the β-blocker Propranolol in 3D Intestinal Fish Model [J].
Langan, Laura M. ;
Owen, Stewart F. ;
Trznadel, Maciej ;
Dodd, Nicholas J. F. ;
Jackson, Simon K. ;
Purcell, Wendy M. ;
Jha, Awadhesh N. .
FRONTIERS IN PHARMACOLOGY, 2018, 9
[38]   Comparison of regulatory frameworks of environmental risk assessments for human pharmaceuticals in EU, USA, and Canada [J].
Lee, Dongyoung ;
Choi, Kyungho .
SCIENCE OF THE TOTAL ENVIRONMENT, 2019, 671 :1026-1035
[39]   Acute toxicity of carbamazepine to juvenile rainbow trout (Oncorhynchus mykiss): Effects on antioxidant responses, hematological parameters and hepatic EROD [J].
Li, Zhi-Hua ;
Zlabek, Vladimir ;
Velisek, Josef ;
Grabic, Roman ;
Machova, Jana ;
Kolarova, Jitka ;
Li, Ping ;
Randak, Tomas .
ECOTOXICOLOGY AND ENVIRONMENTAL SAFETY, 2011, 74 (03) :319-327
[40]   Bioconcentration and metabolism of ketoconazole and effects on multi-biomarkers in crucian carp (Carassius auratus) [J].
Liu, Jianchao ;
Lu, Guanghua ;
Yang, Haohan ;
Yan, Zhenhua ;
Wang, Yonghua ;
Wang, Peifang .
CHEMOSPHERE, 2016, 150 :145-151