Biophysical Control of Bile Duct Epithelial Morphogenesis in Natural and Synthetic Scaffolds

Abstract

The integration of bile duct epithelial cells (cholangiocytes) in artificial liver culture systems is important in order to generate more physiologically relevant liver models. Understanding the role of the cellular microenvironment on differentiation, physiology, and organogenesis of cholangiocytes into functional biliary tubes is essential for the development of new liver therapies, notably in the field of cholangiophaties. In this study, we investigated the role of natural or synthetic scaffolds on cholangiocytes cyst growth, lumen formation and polarization. We demonstrated that cholangiocyte cyst formation efficiency can be similar between natural and synthetic matrices provided that the mechanical properties of the hydrogels are matched. When using synthetic matrices, we also tried to understand the impact of elasticity, matrix metalloprotease-mediated degradation and integrin ligand density on cyst morphogenesis. We demonstrated that hydrogel stiffness regulates cyst formation. We found that controlling integrin ligand density was key in the establishment of large polarized cysts of cholangiocytes. The mechanism of lumen formation was found to rely on cell self-organization and proliferation. The formed cholangiocyte organoids showed a good MDR1 (multi drug resistance protein) transport activity. Our study highlights the advantages of fully synthetic scaffold as a tool to develop bile duct models.

Production and Characterization of an Integrated Multi-Layer 3D Printed PLGA/GelMA Scaffold Aimed for Bile Duct Restoration and Detection

“…One way to enhance cell adhesion was to use bioactive peptides such as collagen, fibronectin and laminin, which exist in extracellular matrix, which promote cell adhesion. For example, P15, a polypeptide from collagen, can promote cell adhesion and modulate a variety of genes ( Carinci et al, 2004 ; Lucidarme et al, 2004 ; Funfak et al, 2019 ). The peptide has been integrated into a dental implant material and achieved good results.…”

Prospect of in vitro Bile Fluids Collection in Improving Cell-Based Assay of Liver Function

“…Spheroids increase cell density, cell-contact polarity, and culture plasticity, including coculture modulation and ECM inclusion (Soldatow et al, 2013;Ramaiahgari et al, 2014;Fatehullah et al, 2016). Unlike sandwich configurations, liver cultures in spheroid configurations allow multiple canalicular sites, thus maintaining superior phase I and II enzyme activities (Soldatow et al, 2013;Ramaiahgari et al, 2014) with a culture age Ni et al, 2016;Ahmed et al, 2017;Kizawa et al, 2017;Vorrink et al, 2017;Baze et al, 2018;Underhills and Khetani, 2018;Fiorotto et al, 2019 (Continued) Frontiers in Toxicology | www.frontiersin.org Swift et al, 2010;Marion et al, 2012;Fukuda et al, 2014;Bachour-El Azzi et al, 2015;Ni et al, 2016;Yang et al, 2016;Yan et al, 2017;Ying et al, 2018 Tanimizu et al, 2007Tanimizu et al, , 2012Kido et al, 2015;Sampaziotis et al, 2015;Miura et al, 2018;Rizki-Safitri et al, 2018Funfak et al, 2019;Du et al, 2020;Hafiz et al, 2021 Potentially high Potentially high Katsuda et al, 2013;Vyas et al, 2018;Wu et al, 2019 Integrated multiorgans Liver-intestine model, multiorganoid chip system Human High d -* Potentially high Maschmeyer et al, 2015;Chen et al, 2017;Choe et al, 2017;…”

Tissue-Engineered Bile Ducts for Disease Modeling and Therapy

“…121 Scaffold stiffness can also directly influence cholangiocyte behavior, 110,122,123 for example, cyst diameter and formation frequency of cholangioids are significantly induced by soft stiffness matrices. 124 In addition, biological properties of materials are equally important, as shown for proliferation and differentiation of a human hepatic cell line on a nanofibrillar cellulose hydrogel. 125 Natural materials have superb biocompatibility to stimulate cell adhesion and growth, however, their use is limited by physical and mechanical stability.…”