Constructing vascularized hepatic tissue by cell-assembled viscous tissue sedimentation method and its application for vascular toxicity assessment. (1st March 2022)

• Record Type: Journal Article
• Authors: Naito, Yasuyuki; Yoshinouchi, Yuka; Sorayama, Yukiko; Kohara, Hiroshi; Kitano, Shiro; Irie, Shinji; Matsusaki, Michiya
• Is Part Of: Acta biomaterialia. Volume 140(2022)
• Page Start: 275
• Page End: 288
• Publication Date: 2022-03-01
• Subjects: 3D Culture -- Heparin -- Collagen -- Hepatic tissue -- DILI; 2D two-dimensional -- 3D three-dimensional -- CAViTs cell assembled viscous tissue by sedimentation -- CD31 platelet endothelial cell adhesion molecule, -- CLSM confocal laser scanning microscope -- DILI drug induced liver injury -- DEG differentially expressed gene -- DMSO dimethyl sulfoxide -- ECGS endothelial cell growth supplement -- ECM extra cellular matrix -- FITC fluorescein isothiocyanate -- LX-2 human hepatic stellate cell line -- MRP2 multidrug resistance-associated protein 2 -- PA pyrrolizidine alkaloid -- PCA principal component analysis -- PBS phosphate-buffered saline -- RT room temperature -- SEC sinusoidal endothelial cell -- SOS sinusoidal obstruction syndrome -- S.D. standard deviation -- VHT vascularized hepatic tissue; Biomedical materials -- Periodicals; 610.28
• DOI: 10.1016/j.actbio.2021.11.027
• Languages: English
• ISSNs: 1742-7061

Abstract

Abstract: In vitro Construction of the liver sinusoidal structure using artificial tissue is an important but worthwhile challenge, particularly for assessing the risk of diseases such as sinusoidal obstruction syndrome (SOS). Current models are unsuitable for evaluating the toxicity because of lacking sinusoidal capillary. In this study, we developed a vascularized hepatic tissue (VHT) using a unique tissue engineering technique, the cell assembled viscous tissue by sedimentation (CAViTs) method. The "viscous bodies" created using the CAViTs method exhibited significant self-assembly within 6 h after seeding, promoting cell-cell interaction. The level of albumin secreted by the VHT was four times higher than that of 2D-coculture and maintained for 1 month. The gene expression pattern of the VHT was closer to that of total human liver, compared with the 2D system. Quantitative evaluations of the vascular structure of VHT treated with two typical SOS-inducing compounds, monocrotaline and retrorsine, revealed higher sensitivity (IC50 = 40.35 µM), 19.92 times higher than the cell-viability assay. Thus, VHT represents an innovative in vitro model that mimics the vessel network structure and could become a useful tool for the early screening of compounds associated with a risk of vascular toxicity. Statement of significance: Mimicking sinusoidal structures in in vitro liver model is important to consider from the perspective of predicting hepatotoxicity such like sinusoidal obstruction syndrome (SOS). However, it was difficult to reconstruct the vascular structure within the hepatocyte-rich environment. In this study, we constructed a vascularized hepatic tissue in a high-throughput manner by a unique method using collagen and heparin, and evaluated its applicability to toxicity assessment. Vessel morphology analysis of the model treated by monocrotaline, which is a well-known SOS-inducing compound, could predict the toxicity with higher sensitivity. To the best of our knowledge, this is the first report to provide vascularized hepatic tissues using sinusoidal endothelial cells at least for demonstrating applicability to the evaluation of SOS induction risk. Graphical abstract: Image, graphical abstract