Hepatocyte spheroid culture on fibrous scaffolds with grafted functional ligands as an in vitro model for predicting drug metabolism and hepatotoxicity.
Hepatocyte spheroid culture on fibrous scaffolds with grafted functional ligands as an in vitro model for predicting drug metabolism and hepatotoxicity.
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DOI：
10.1016/j.actbio.2015.09.027
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发表时间：
2015-12
影响因子：
9.7
通讯作者：
Shili Yan;Jiaojun Wei;Yaowen Liu;Hong Zhang;Jianmei Chen;Xiaohong Li
中科院分区：
文献类型：
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作者：
Shili Yan;Jiaojun Wei;Yaowen Liu;Hong Zhang;Jianmei Chen;Xiaohong Li
鉴定维持肝细胞功能的生物底物对于为体外药物筛选提供可靠和可预测的模型至关重要。在目前的研究中，肝细胞在高多孔纤维支架上建立了三维培养，并嫁接了半乳糖和RGD，以提供广泛的细胞-细胞和细胞-支架在空间上的相互作用。孔径和配体密度对肝细胞球体的形成有显著影响，在纤维支架上平衡细胞的保留、粘附和迁移。平均孔径为60 μm、半乳糖表面接枝密度为5.9 nmol/cm2、RGD为6.9 pmol/cm2的纤维支架为肝细胞浸润和多细胞球体形成提供了最佳微环境。白蛋白和尿素的合成以及I期酶（CYP 3A11和CYP 2C9）和II期酶的活性也得到了显著的促进。肝细胞在最佳支架上对睾酮和对乙酰氨基酚的体外代谢试验表明，预测清除率分别为50.7和22.6 ml/min/kg，与大鼠体内清除率相当。盐酸胺碘酮和对乙酰氨基酚的体外肝毒性试验预测了半最大有效浓度（EC50），反映了它们在人体内毒性的血浆浓度。此外，最佳支架上肝细胞的酶活性、预测清除率和肝毒性值对CYP 3A11和II期酶的特异性诱导剂或抑制剂具有敏感的反应性，在一定程度上表现出体内外相关性。这些结果表明，在纤维支架上培养肝细胞球形细胞是一种潜在的体外检测模型，可以预测其体内药物代谢、肝毒性和药物-药物相互作用。
The identification of a biologic substrate for maintaining hepatocyte functions is essential to provide reliable and predictable models forin vitrodrug screening. In the current study, a three-dimensional culture of hepatocytes was established on highly porous fibrous scaffolds with grafted galactose and RGD to afford extensive cell–cell and cell–scaffold interactions spatially. The pore size and ligand densities indicated significant effects on the formation of hepatocyte spheroids in balancing the cell retention, adhesion, and migration on fibrous scaffolds. Fibrous scaffolds with an average pore size of 60 μm and surface grafting densities of galactose at 5.9 nmol/cm2and RGD at 6.9 pmol/cm2provided optimal microenvironments for hepatocyte infiltration and multicellular spheroid formation. Significant promotions were also demonstrated in the syntheses of albumin and urea and the activities of phase I (CYP 3A11 and CYP 2C9) and phase II enzymes. Thein vitrometabolism tests on testosterone and acetaminophen by hepatocytes on the optimal scaffolds indicated the predicated clearance rates of 50.7 and 22.6 ml/min/kg, respectively, which were comparable to thein vivovalues of rats. Thein vitrohepatotoxicity tests on amiodarone hydrochloride and acetaminophen predicted the half maximal effective concentrations (EC50) to reflect thein vivotoxic plasma concentrations in human. In addition, the enzyme activities, predicted clearance rates and hepatotoxicity values of hepatocytes on the optimal scaffolds experienced sensitive responsiveness to specific inducers or inhibitors of CYP 3A11 and phase II enzymes, exhibitingin vivo–in vitrocorrelations to a certain extent. These results demonstrate the feasibility of hepatocyte spheroid culture on fibrous scaffolds as an potentialin vitrotesting model to predict thein vivodrug metabolism, hepatotoxicity, and drug–drug interactions.