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Compos. Sci."],"abstract":"<jats:p>Osteochondral defects present significant challenges for effective tissue regeneration due to the complex composition of bone and cartilage. To address this challenge, this study presents the fabrication of hierarchical scaffolds combining chitosan\/\u03b2-tricalcium phosphate (\u03b2-TCP) to simulate a bone-like layer, interconnected with a silk fibroin layer to mimic cartilage, thus replicating the cartilage-like layer to mimic the native osteochondral tissue architecture. The scaffolds were produced by freeze-drying and then crosslinking with genipin. They have a crosslinking degree of up to 24%, which promotes a structural rearrangement and improved connection between the different layers. Micro-CT analysis demonstrated that the structures have distinct porosity values on their top layer (up to 84%), interface (up to 65%), and bottom layer (up to 77%) and are dependent on the concentration of \u03b2-tricalcium phosphate used. Both layers were confirmed to be clearly defined by the distribution of the components throughout the constructs, showing adequate mechanical properties for biomedical use. The scaffolds exhibited lower weight loss (up to 7%, 15 days) after enzymatic degradation due to the combined effects of genipin crosslinking and \u03b2-TCP incorporation. In vitro studies showed that the constructs supported ATDC5 chondrocyte-like cells and MC3T3 osteoblast-like cells in duo culture conditions, providing a suitable environment for cell adhesion and proliferation for up to 14 days. Overall, the physicochemical properties and biological results of the developed chitosan\/\u03b2-tricalcium phosphate\/silk fibroin bilayered scaffolds suggest that they may be potential candidates for osteochondral tissue strategies.<\/jats:p>","DOI":"10.3390\/jcs8030085","type":"journal-article","created":{"date-parts":[[2024,2,26]],"date-time":"2024-02-26T11:31:21Z","timestamp":1708947081000},"page":"85","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":9,"title":["Chitosan-Based Hierarchical Scaffolds Crosslinked with Genipin"],"prefix":"10.3390","volume":"8","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-0748-2885","authenticated-orcid":false,"given":"Lya","family":"Piaia","sequence":"first","affiliation":[{"name":"Laboratory of Innovation on Additive Manufacturing and Molding (NIMMA), Department of Mechanical Engineering (EMC), Federal University of Santa Catarina (UFSC), Florian\u00f3polis 88040-900, SC, 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M.","family":"Fernandes","sequence":"additional","affiliation":[{"name":"3B\u2019s Research Group, I3Bs\u2014Research Institute on Biomaterials, Biodegradables and Biomimetics, University of Minho, Headquarters of the European Institute of Excellence on Tissue Engineering and Regenerative Medicine, AvePark, Parque de Ci\u00eancia e Tecnologia, Zona Industrial da Gandra, 4805-017 Barco, Guimar\u00e3es, Portugal"},{"name":"ICVS\/3B\u2019s\u2014PT Government Associate Laboratory, 4710-057 Braga, Guimar\u00e3es, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6951-0194","authenticated-orcid":false,"given":"Joana M.","family":"Gomes","sequence":"additional","affiliation":[{"name":"3B\u2019s Research Group, I3Bs\u2014Research Institute on Biomaterials, Biodegradables and Biomimetics, University of Minho, Headquarters of the European Institute of Excellence on Tissue Engineering and Regenerative Medicine, AvePark, Parque de Ci\u00eancia e Tecnologia, Zona Industrial da Gandra, 4805-017 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