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Thiago Domingues Stocco,1– 3 Mayara Cristina Moreira Silva,4 Marcus Alexandre Finzi Corat,4 Gabriely Gonçalves Lima,5 Anderson Oliveira Lobo5 1Faculty of Medical Sciences, Unicamp - State University of Campinas, Campinas, SP, Brazil; 2Division of Engineering in Medicine, Department of Medicine, Brigham and Women’s Hospital, Harvard Medical School, Cambridge, MA, USA; 3UNISA - University of Santo Amaro, São Paulo, Brazil; 4Multidisciplinary Center for Biological Research, Unicamp – State University of Campinas, Campinas, SP, Brazil; 5LIMAV—Interdisciplinary Laboratory for Advanced Materials, BioMatLab, UFPI—Federal University of Piauí, Teresina, PI, BrazilCorrespondence: Anderson Oliveira Lobo, Email lobo@ufpi.edu.brIntroduction: Three of the main requirements that remain major challenges in tissue engineering of the knee meniscus are to engineer scaffolds with compatible anatomical shape, good mechanical properties, and microstructure able to mimic the architecture of the extracellular matrix (ECM). In this context, we presented a new biofabrication strategy to develop a three-dimensional (3D) meniscus-regenerative scaffold with custom-made macroscopic size and microarchitecture bioinspired by the organization of structural fibers of native tissue ECM.Methods: The concept was based on the combination of bioprinted cell-laden hydrogel (type 1 collagen) reinforced by multilayers of biomimetically aligned electrospun nanofibrous mats (polycaprolactone/carbon nanotubes, PCL/CNT), using a patient-specific 3D digital meniscus model reconstructed from MRI data by free and open-source software.Results: The results showed that the incorporation of aligned nanofibers sheets between the hydrogel layers enhanced the scaffold’s structural integrity and shape fidelity compared to the nanofiber-free collagen hydrogel. Furthermore, mechanical compression tests demonstrated that the presence of nanofiber layers significantly improved the mechanical properties of the bioprinted construct. Importantly, the introduction of PCL/CNT nanofibrous mats between the layers of the bioprinted collagen hydrogel did not negatively affect cell viability, in which mesenchymal stem cells remained viable even after 7 days of culture within the scaffold.Conclusion: Overall, these findings evidence that this bioengineering approach offers a promising strategy for fabricating biomimetic meniscus scaffolds for tissue engineering.Keywords: tissue engineering, regenerative medicine, meniscus, biomaterials, nanotechnology, 3D printing