Novel 3D nano-fabricated zonal scaffolds for articular cartilage regeneration

Event Abstract Back to Event Novel 3D nano-fabricated zonal scaffolds for articular cartilage regeneration Ying Yang1, Hamza Owida1 and Nicola Kuiper1 1 Keele University, Institute of Science and Technology in Medicine, United Kingdom Introduction: Articular cartilage is a thin hydrated tissue, which covers articulating surfaces. It has an anisotropic, zone-specific structure, extending from the articular surface to the subchondral bone[1]. The extracellular matrix (ECM) is organised into distinct zones; superficial, middle and deep. As a result of differences in chondrocyte activity and mechanical loading, the ECM composition and its structure varies greatly between each zone[2]. The main challenge associated with cartilage tissue engineering is to generate functional tissue with zonal organisation similarity to the native articular cartilage tissue. The previous zonal specific cartilage studies did not generate distinct cellular orientation and ECM variation in the corresponding zones as in native counterpart. The aim of this project is to fabricate and utilize nanofibrous scaffolds with different fiber organizations to mimic the native variations of ECM between zones, enabling better cartilage tissue formation. Methods: Articular cartilage was dissected from the femoral condyle joints of cows within 3 h of slaughter. Chondrocytes were released from their ECM and expanded in monolayer culture. Zonal-specific nanofibrous scaffolds were produced by electrospinning with a modified protocol previously described[3]. Novel collectors were developed to generate the respective nanofiber scaffold able to induce cell and ECM organization and orientation in superficial, middle and deep zones. The 100,000 chondrocytes per scaffold were seeded in the three types of nanofiber scaffolds encapsulated by agarose hydrogel respectively. This study adapted a sandwich construction with two layers of agarose to deliver and maintain nanofibers’ orientation. The bottom agarose layer restricted cells attaching and orientating on nanofiber meshes, whilst the top layer of agarose provided the support necessary to stabilize the orientation of nanofibers meshes and cells’. Various characterisation assays including Toluidine blue staining to assess glycosaminoglycans (GAG) distribution, PicoGreen assay to assess DNA content, and DMMB assay to quantify sulphated GAG content have been undertaken. Results and Discussion: Nanofibrous scaffolds with aligned fiber (for superficial zone), random fiber (for middle zone) and bundle-like fiber arrangement (for deep zone) have been produced. The seeded chondrocytes on nanofiber scaffolds have shown different morphologies and GAG contents (Figure 1), normalisation of GAG content per DNA content (Figure 2) has shown variation between different 3D zonal scaffolds. Chondrocytes in the superficial zone scaffold exhibited elongated, low GAG but lower DNA content; whilst deep zone scaffold produced aligned chondrocyte clusters with high GAG content and low DNA amount; middle zone scaffold had the cellular behaviour between both of other scaffolds. Figure 1: Toluidine blue staining of chondrocytes grown on different nano-fabricated 3D zonal scaffolds. 10X. Figure 2: Normalisation of GAG content per DNA content and DNA content in the different nano-fabricated 3D zonal scaffolds Conclusions: Nano-fabricated zonal-specific 3D scaffolds have induced distinct chondrogenic potential, which matches the cellular alignment and ECM content variation in native cartilage. This new approach demonstrates the hope to generate structural and functional cartilage tissue similar to native counterpart. References:[1] Buckwalter, J. A., & Mankin, H. J. (1997). Instructional course lectures, 47, 477-486.[2] Klein, T. J., Malda, J., Sah, R. L., et al. (2009). Tissue Engineering Part B: Reviews, 15(2), 143-.157.[3] Yang, Y., Wimpenny, I., & Ahearne, M. (2011). Nanomedicine: Nanotechnology, Biology and Medicine, 7(2), 131-136 Keywords: nanofiber, 3D scaffold Conference: 10th World Biomaterials Congress, Montréal, Canada, 17 May - 22 May, 2016. Presentation Type: General Session Oral Topic: Biomaterials in constructing tissue substitutes Citation: Yang Y, Owida H and Kuiper N (2016). Novel 3D nano-fabricated zonal scaffolds for articular cartilage regeneration. Front. Bioeng. Biotechnol. Conference Abstract: 10th World Biomaterials Congress. doi: 10.3389/conf.FBIOE.2016.01.02967 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 Ying Yang Hamza Owida Nicola Kuiper Google Ying Yang Hamza Owida Nicola Kuiper Google Scholar Ying Yang Hamza Owida Nicola Kuiper PubMed Ying Yang Hamza Owida Nicola Kuiper 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.