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• W2327094310 abstract "Event Abstract Back to Event Tissue engineering cell-based bioelectronics Ulises Aregueta Robles1, Khoon S. Lim2, Penny J. Martens1, Nigel H. Lovell1, Laura A. Poole-Warren1 and Rylie A. Green1 1 University of NSW, Graduate School of Biomedical Engineering, Australia 2 University of Otago, Department of Orthopaedic Surgery & Musculoskeletal Medicine, New Zealand Tissue engineered electrodes aim to deliver a living cellular layer to interface bionic devices with target tissue by establishing synaptic connections between the electrode and target nerve. A significant challenge in developing this technology is to engineer a hydrogel carrier that can support growth and function of complex neuronal networks. To provide an appropriate neural cell niche, it is desirable to have not only neural cells, but supporting glia within the hydrogel carrier. It has been hypothesized that a hydrogel tailored to support glial cell survival and function will be able to support neuronal development. Poly(vinyl alcohol) functionalized with tyramine (PVA-Tyr) is a hydrogel system that supports covalent incorporation of non-modified tyrosine rich proteins, mimicking the biological environment and enabling 3D cell encapsulation[1]. PVA-Tyr has been co-polymerized with gelatin and sericin and the resulting gel supported glial cell growth. However, development of an appropriate system for use at the neural interface requires tailoring of the hydrogel properties such that (i) the mechanical stiffness is similar to that of neural tissues, and (ii) the hydrogel degradation rate is matched to cell maturation and development of extracellular matrix (ECM). Since physical and mechanical properties of PVA relate to the percent polymer[2], the aim of this research was to investigate the impact of varying the hydrogel macromer percentage on the mechanical stiffness and physical degradation of PVA-Tyr/sericin/gelatin (PVA-T/S/G), and the subsequent impact on glial cell survival and development. Two variants of PVA-T/S/G, 10 wt% and 5 wt%, were developed and used to encapsulate peripheral glia (Schwann cells). PVA-Tyr (8 or 3 wt%) hydrogel was crosslinked with gelatin (1 wt%) and sericin (1 wt%) as previously described[1]. Schwann cells (SC, SCL4.1/F7) were encapsulated at 107 cells/mL. Cellular viability was tracked by live/dead staining. Hydrogel stiffness was measured by compression tests (50 N load cell, 0.5 mm/min, 5 - 15% strain). Cells were immunostained for S100 (mature SC marker), laminin and collagen-IV (ECM deposition) and bisbenzimide (nuclei). Hydrogel degradation was tracked to the point of reverse gelation. All the tests were performed at 0, 1, 5 and 10 days. Cells remained viable at all the time points. Compression tests showed that PVA-T/S/G at 10 wt% had a mean compressive modulus of 41.5 kPa at Day 0, which reduced to 0.5 kPa by Day 10. The corresponding compressive modulus for the 5 wt% hydrogel was 6.1 kPa at Day 0 and 0.6 kPa at Day 10. SCs interact preferably with substrates having a stiffness from 1 - 10 kPa[3], where they adopt bipolar morphologies that support nerve guidance and repair in vivo[4]. In both systems at 5 days post-encapsulation bipolar morphologies were evident (see Figure 1), but were more prolific in the 5 wt% hydrogels. Presence of laminin and collagen-IV was also observed by immunostaining. Expression of these molecules is an indication of mature and functional SCs[5]. Both systems were shown to be completely degraded at 15 ± 3 days. While these different hydrogels provide similar support to SCs the ability to vary the hydrogel stiffness is expected to be key to the development of a complex living electrode with both glia and neuronal cell types, as previously proposed[6]. Figure 1. SC embedded in 10% (left) and 5%(right) PVA-T/S/G hydrogel at 5 days in culture. Nuclei (blue). Collagen-IV (green). Scale bar = 50 μm SCs were able to adopt bipolar morphologies and express ECM required for neuronal development with 5 and 10 wt% hydrogels. These outcomes suggest that the PVA-T/S/G hydrogels provide physical and biochemical support for SC survival and maturation. Future studies will assess the capacity of SCs to support neuronal survival within PVA-T/S/G. Australian Research Council (ARC) through its Special Research Initiative (SRI) in Bionic Vision Science and Technology grant to Bionic Vision Australia (BVA)References:[1] Lim, K.S., et al., Covalent incorporation of non-chemically modified gelatin into degradable PVA-tyramine hydrogels. Biomaterials, 2013. 34(29): p. 7097-7105[2] Martens, P., et al., Effect of Poly(vinyl alcohol) Macromer Chemistry and Chain Interactions on Hydrogel Mechanical Properties. Chemistry of Materials, 2007. 19(10): p. 2641-2648.[3] Lacour, S.P., et al., Flexible and stretchable micro-electrodes for in vitro and in vivo neural interfaces. Medical & Biological Engineering & Computing, 2010. 48(10): p. 945-954[4] Fu, S. and T. Gordon, The cellular and molecular basis of peripheral nerve regeneration. Molecular Neurobiology, 1997. 14(1-2): p. 67-116[5] Chernousov, M.A., et al., Regulation of Schwann cell function by the extracellular matrix. Glia, 2008. 56(14): p. 1498-1507[6] Aregueta-Robles, U.A., et al., Producing 3D Neuronal Networks in Hydrogels for Living Bionic Device Interfaces. Proceedings of the 37th Annual International Conference of the IEEE Engineering in Medicine and Biology Society, 2015 Keywords: Hydrogel, Tissue Engineering, 3D scaffold, Polymeric material Conference: 10th World Biomaterials Congress, Montréal, Canada, 17 May - 22 May, 2016. Presentation Type: Poster Topic: Biomaterials in nerve regeneration Citation: Aregueta Robles U, Lim KS, Martens PJ, Lovell NH, Poole-Warren LA and Green RA (2016). Tissue engineering cell-based bioelectronics. Front. Bioeng. Biotechnol. Conference Abstract: 10th World Biomaterials Congress. doi: 10.3389/conf.FBIOE.2016.01.02811 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. Login Required This action requires you to be registered with Frontiers and logged in. To register or login click here. Abstract Info Abstract The Authors in Frontiers Ulises Aregueta Robles Khoon S Lim Penny J Martens Nigel H Lovell Laura A Poole-Warren Rylie A Green Google Ulises Aregueta Robles Khoon S Lim Penny J Martens Nigel H Lovell Laura A Poole-Warren Rylie A Green Google Scholar Ulises Aregueta Robles Khoon S Lim Penny J Martens Nigel H Lovell Laura A Poole-Warren Rylie A Green PubMed Ulises Aregueta Robles Khoon S Lim Penny J Martens Nigel H Lovell Laura A Poole-Warren Rylie A Green Related Article in Frontiers Google Scholar PubMed Abstract Close Back to top Javascript is disabled. Please enable Javascript in your browser settings in order to see all the content on this page." @default.
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