A composite Gelatin/hyaluronic acid hydrogel as an ECM mimic for developing mesenchymal stem cell derived epithelial tissue patches

Kumar, Pramod; Ciftci, Sait; Barthes, Julien; Knopf-Marques, Helena; Muller, C�line Blandine; Debry, Christian; Vrana, Nihal E.; Ghaemmaghami, Amir M.

Abstract

Here we report fabrication of Gelatin based biocomposite films and their application in developing epithelial patches. The films were loaded with an epithelial cell growth factor cocktail and used as an extracellular matrix mimic (ECM mimic) for in vitro regeneration of organised respiratory epithelium using Calu‐3 cell line and mesenchymal stem cells (MSCs). Our data show differentiation of Calu‐3 cells on composite films as evidenced by tight junction protein expression and barrier formation. The films also supported attachment, migration and proliferation of alveolar basal epithelial cell line A549. We also show the suitability of the composite films as a biomimetic scaffold and growth factor delivery platform for differentiation of human MSCs to epithelial cells. MSCs differentiation to the epithelial lineage was confirmed by staining for epithelial and stem cell specific markers. Our data show that the MSCs acquire the epithelial characteristics after two weeks with significant reduction in vimentin, increase in pan cytokeratin expression as well as morphological changes. However, despite the expression of epithelial lineage markers these cells did not form fully functional tight junctions as evidenced by low expression of junctional protein ZO1. Further optimisation of culture conditions and growth factor cocktail is required to enhance tight junction formation in MSCs derived epithelial cells on the composite hydrogels. Nevertheless, our data clearly highlight the possibility of using MSCs in epithelial tissue engineering and the applicability of the composite hydrogels as transferrable ECM mimics and delivery platforms with potential applications in regenerative medicine and in vitro modelling of barrier tissues.

• Journal Article Type: Article
• Acceptance Date: Sep 4, 2019
• Online Publication Date: Oct 9, 2019
• Publication Date: Jan 1, 2020
• Deposit Date: Sep 18, 2019
• Publicly Available Date: Oct 10, 2020
• Journal: Journal of Tissue Engineering and Regenerative Medicine
• Print ISSN: 1932-6254
• Electronic ISSN: 1932-7005
• Publisher: Wiley
• Peer Reviewed: Peer Reviewed
• Volume: 14
• Issue: 1
• Pages: 45-57
• DOI: https://doi.org/10.1002/term.2962
• Keywords: Mesenchymal stem cell, Gelatin/HA, Hydrogel, Growth factors, Controlled release, Epithelium differentiation
• Public URL: https://nottingham-repository.worktribe.com/output/2630427
• Publisher URL: https://onlinelibrary.wiley.com/doi/abs/10.1002/term.2962
• Additional Information: Received: 2018-06-18; Accepted: 2019-09-04; Published: 2019-10-09