Event Abstract Back to Event Layer-by-layer assembly of collagen and fibronectin for tissue engineering applications Sara Mauquoy1 and Christine Dupont-Gillain1 1 Université Catholique de Louvain, Institute of Condensed Matter and Nanosciences, Belgium Introduction: In vivo, cells are embedded in an extracellular matrix (ECM). Collagen (Col) and fibronectin (Fn), both involved in signalling from cells to the ECM and conversely, are two major components of the ECM. Tailoring biointerfaces based on these two biomolecules will improve the control of cell-material interactions. Layer-by-layer (LbL) assembly, based on the alternate surface dipping in a polycation and a polyanion solution, is a versatile approach to design structures with a vertical heterogeneity. Assembling proteins by LbL is however challenging in reason of their polyampholyte character and their particular structure. The mechanism of assembly may moreover rely on specific interactions. Here, we aim at combining Fn and Col in LbL assemblies to provide a biomimetic environment to stem cells. Methods: LbL assembly of Fn and native or denatured Col is performed on a polystyrene (PS) substrate from different buffers. A poly(ethyleneimine) (PEI) anchoring layer is sometimes added prior to the assembly. LbL assemblies are also compared to layers obtained by simultaneous adsorption of Fn and Col (Fn+Col). Quartz crystal microbalance is used to determine the wet deposited mass, ellipsometry to measure the dry thickness of the films, contact angle measurements to characterize their wettability and AFM to examine their morphology. Adhesion, proliferation (Alamar blue assay) and differentiation (Alizarin red staining) experiments are performed with adipose-derived mesenchymal stem cells. Results and Discussion: Results show that LbL assembly is generally better with denatured compared to native Col, and that thicker films are obtained when using PEI. The buffer also influences the assembly, with HEPES buffer giving the best results. The combination of results from all techniques indicates successful build-up of multilayers, with a growth which is however not sustainable. The comparison of films obtained by LbL assembly and simultaneous adsorption reveals differences in terms of wettability and organization, which could trigger different cell responses. Cell studies are made on films made of Fn and native Col in HEPES buffer. Proliferation data (Fig 1) show that PEI decreases cell growth. However, the metabolic activity reaches the same level with or without PEI after 13 days. PEI is thus probably cytotoxic in the beginning of grow phase. All the protein films improve proliferation but differences between deposition methods are not evidenced. Differentiation study shows an influence of the deposition method, with LbL deposition in absence of PEI giving poor differentiation compared to other films. Interestingly, films constructed on PEI-coated PS allow a better differentiation, especially for LbL deposition (Fig 2). Conclusions: The deposition of Col and Fn in a variety of conditions is now better understood and influences cell behavior. More tests will be performed to study in detail the relation between film properties and cell differentiation. S. M. thanks the Belgian National Foundation for Scientific Research (FNRS) for her Research Fellow position and for the financial support. Keywords: Extracellular Matrix, Tissue Engineering, stem cell, bioinerface Conference: 10th World Biomaterials Congress, Montréal, Canada, 17 May - 22 May, 2016. Presentation Type: Poster Topic: Layer-by-layer deposition techniques Citation: Mauquoy S and Dupont-Gillain C (2016). Layer-by-layer assembly of collagen and fibronectin for tissue engineering applications. Front. Bioeng. Biotechnol. Conference Abstract: 10th World Biomaterials Congress. doi: 10.3389/conf.FBIOE.2016.01.02742 Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters. The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated. Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed. For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions. Received: 27 Mar 2016; Published Online: 30 Mar 2016. 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