@article{discovery10136333, month = {September}, volume = {13}, number = {19}, note = {{\copyright} 2021 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https:// creativecommons.org/licenses/by/ 4.0/).}, year = {2021}, title = {Optimization and validation of a novel three-dimensional co-culture system in decellularized human liver scaffold for the study of liver fibrosis and cancer}, journal = {Cancers}, keywords = {decellularized liver scaffolds; drug screening; liver fibrosis; liver cancer; sorafenib; regorafenib; STAT3; SHP-1; TGF{\ensuremath{\beta}}1; EMT; E-cadherin; 3D in vitro model}, url = {https://doi.org/10.3390/cancers13194936}, abstract = {The introduction of new preclinical models for in vitro drug discovery and testing based on 3D tissue-specific extracellular matrix (ECM) is very much awaited. This study was aimed at developing and validating a co-culture model using decellularized human liver 3D ECM scaffolds as a platform for anti-fibrotic and anti-cancer drug testing. Decellularized 3D scaffolds obtained from healthy and cirrhotic human livers were bioengineered with LX2 and HEPG2 as single and co-cultures for up to 13 days and validated as a new drug-testing platform. Pro-fibrogenic markers and cancer phenotypic gene/protein expression and secretion were differently affected when single and co-cultures were exposed to TGF-{\ensuremath{\beta}}1 with specific ECM-dependent effects. The anti-fibrotic efficacy of Sorafenib significantly reduced TGF-{\ensuremath{\beta}}1-induced pro-fibrogenic effects, which coincided with a downregulation of STAT3 phosphorylation. The anti-cancer efficacy of Regorafenib was significantly reduced in 3D bioengineered cells when compared to 2D cultures and dose-dependently associated with cell apoptosis by cleaved PARP-1 activation and P-STAT3 inhibition. Regorafenib re-versed TGF-{\ensuremath{\beta}}1-induced P-STAT3 and SHP-1 through induction of epithelial mesenchymal marker E-cadherin and downregulation of vimentin protein expression in both co-cultures engrafting healthy and cirrhotic 3D scaffolds. In their complex, the results of the study suggest that this newly proposed 3D co-culture platform is able to reproduce the natural physio-pathological microenvi-ronment and could be employed for anti-fibrotic and anti-HCC drug screening.}, issn = {2072-6694}, author = {Thanapirom, K and Caon, E and Papatheodoridi, M and Frenguelli, L and Al-akkad, W and Zhenzhen, Z and Vilia, MG and Pinzani, M and Mazza, G and Rombouts, K} }