Development of a hyaluronic acid/plasma-derived fibrin hydrogel for the optimization of dermo-epidermal autologous equivalents

Event Abstract Back to Event Development of a hyaluronic acid/plasma-derived fibrin hydrogel for the optimization of dermo-epidermal autologous equivalents Jose Manuel Rey Legidos1, Nieves Cubo Mateo1, Andres Montero Simon1, Marta Garcia Diez1, 2, 3, Diego Velasco Bayón1, 2 and Jose Luis Jorcano Noval1, 4 1 Universidad Carlos III de Madrid, Bioengineering and Space Engineering, Spain 2 Instituto de Investigación Sanitaria de la Fundación Jiménez Díaz, Spain 3 CIEMAT-CIBER de Enfermedades Raras (U714, Spain 4 Regenerative Medicine Unit and Epithelial Biomedicine Division, CIEMAT, Spain Introduction: Over the past several years, the generation of a human plasma-derived fibrin bilayer (including dermis and epidermis) skin model by our group has been applied successfully to treat burns, traumatic and surgical wounds[1]. However, limitations of plasma-derived fibrin hydrogels such as low mechanical properties[2], high degradation rates and shrinking during in vitro culture or implantation necessitate the development of new approaches to address these needs. The combination of plasma-derived fibrin with other natural polymers can be considered an interesting possibility for improving the hydrogel mechanical properties without compromising biocompatibility. Hyaluronic acid (HA) is a natural polymer which is present in the extracellular matrix (ECM) of the skin (estimated 0.12-0.3% w/v) and it is also a contributor of dermal turgor. In the present work, we incorporated a thiolated form of HA crosslinked with poly(ethylene glycol) diacrylate (PEGDA) in a well-established protocol for producing plasma-derived fibrin dermal equivalents to improve their biological and mechanical properties. Materials and Methods: Plasma-derived fibrin hydrogels were prepared following the protocol described previously by our group1. In this sense, 883 μl of human plasma (2.21 mg/ml of fibrinogen) was diluted in 600 μl of saline solution (0.9 % w/v). For crosslinking, 130 μl of a CaCl2 (1% w/v) solution was added. Then the protocol was modified to incorporate thiolated HA and PEGDA at concentrations of 0.1-0.2% and 0-0.05% (w/v) respectively, while concentrations of the rest of the reagents were kept constant in all hydrogels to comply with the above mentioned protocol. Formation of gels and gelation times were assessed using flip-flop method and rheology measurements. The swelling/deswelling studies of HA-fibrin hydrogels were performed. Human fibroblasts were embedded in the hydrogels following the same protocol and toxicity tests were performed. The capacity of the human fibroblasts to contract the hydrogels was also studied at 0, 3, 7 and 10 days. Results and Discussion: Introduction of HA and PEGDA in fibrin hydrogels increased the gelation time to 16-35 minutes (with the exception of HA 0.2 % and PEGDA 0.05%) due to possible interactions between HA, PEGDA and fibrinogen respectively (Figure1). It was observed that HA-fibrin hydrogels retained shape and bulk integrity in contrast to fibrin hydrogels. Fibrin gels were confirmed to have strong contractile behaviour, while HA-fibrin hydrogels inhibited fibrin contraction (Figure2). Introduction of HA and PEGDA in fibrin gels did not affect the viability of the human fibroblasts. All hydrogels with human fibroblasts embedded contracted after 10 days, but HA-fibrin gels contracted significantly less than control fibrin gels. HA-fibrin gels with PEGDA enhanced very significantly fibroblast proliferation compared with fibrin hydrogels and HA-fibrin hydrogels without PEGDA at day 7 (Figure3). Conclusion: In this work, it was demonstrated that the incorporation of HA and PEGDA to plasma-derived fibrin hydrogels increased the mechanical properties and inhibited the fibrin hydrogel contraction with and without cells. It was also confirmed a higher proliferation of human fibroblasts when using HA-fibrin hydrogels with PEGDA. Further in vitro and in vivo experiments are required to assess the quality of the engineered skin. The authors acknowledge financial support of this work to the Universidad Carlos III de Madrid, and the plasma supply by the Centro Comunitario de Sangre y Tejidos in AsturiasReferences:[1] Llames S.G., et al.Transplantation. (2004); 77:350-355.[2] Haugh, M.G., et al. J. Mech. Behav. Biomed. Mater. (2012); 16:66-72. Keywords: Hydrogel, biomaterial, Tissue Regeneration, Burn wound Conference: 10th World Biomaterials Congress, Montréal, Canada, 17 May - 22 May, 2016. Presentation Type: General Session Oral Topic: Biomaterials in constructing tissue substitutes Citation: Rey Legidos J, Cubo Mateo N, Montero Simon A, Garcia Diez M, Velasco Bayón D and Jorcano Noval J (2016). Development of a hyaluronic acid/plasma-derived fibrin hydrogel for the optimization of dermo-epidermal autologous equivalents. Front. Bioeng. Biotechnol. Conference Abstract: 10th World Biomaterials Congress. doi: 10.3389/conf.FBIOE.2016.01.00743 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 Jose Manuel Rey Legidos Nieves Cubo Mateo Andres Montero Simon Marta Garcia Diez Diego Velasco Bayón Jose Luis Jorcano Noval Google Jose Manuel Rey Legidos Nieves Cubo Mateo Andres Montero Simon Marta Garcia Diez Diego Velasco Bayón Jose Luis Jorcano Noval Google Scholar Jose Manuel Rey Legidos Nieves Cubo Mateo Andres Montero Simon Marta Garcia Diez Diego Velasco Bayón Jose Luis Jorcano Noval PubMed Jose Manuel Rey Legidos Nieves Cubo Mateo Andres Montero Simon Marta Garcia Diez Diego Velasco Bayón Jose Luis Jorcano Noval 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.