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Indirect printing of hierarchical patient-specific scaffolds for meniscus tissue engineering

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Abstract

The complex meniscus tissue plays a critical role in the knee. The high susceptibility to injury has led to an intense pursuit for better tissue engineering regenerative strategies, where scaffolds play a major role. In this study, indirect printed hierarchical multilayered scaffolds composed by a silk fibroin (SF) upper layer and an 80/20 (w/w) ratio of SF/ionic-doped β-tricalcium phosphate (TCP) bottom layer were developed. Furthermore, a comparative analysis between two types of scaffolds produced using different SF concentrations, i.e., 8% (w/v) (Hi8) and 16% (w/v) (Hi16) was performed. In terms of architecture and morphology, the produced scaffolds presented homogeneous porosity in both layers and no differences were observed when comparing both scaffolds. A decrease in terms of mechanical performance of the scaffolds was observed when SF concentration decreased from 16 to 8% (w/v). Hi16 revealed a static compressive modulus of 0.66 ± 0.05 MPa and dynamical mechanical properties ranging from 2.17 ± 0.25 to 3.19 ± 0.38 MPa. By its turn, Hi8 presented a compressive modulus of 0.27 ± 0.08 MPa and dynamical mechanical properties ranging from 1.03 ± 0.08 MPa to 1.56 ± 0.13 MPa. In vitro bioactivity studies showed formation of apatite crystals onto the surface of Hi8 and Hi16 bottom layers. Human meniscus cells (hMCs) and human primary osteoblasts were cultured separately onto the top layer (SF8 and SF16) and bottom layer (SF8/TCP and SF16/TCP) of the hierarchical scaffolds Hi8 and Hi16, respectively. Both cell types showed good adhesion and proliferation as denoted by the live/dead staining, Alamar Blue assay and DNA quantification analysis. Subcutaneous implantation in mice revealed weak inflammation and scaffold’s integrity. The hierarchical indirect printed SF scaffolds can be promising candidate for meniscus TE scaffolding applications due their suitable mechanical properties, good biological performance and possibility of being applied in a patient-specific approach.

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Acknowledgements

The authors thank the financial support provided by the Portuguese Foundation for Science and Technology (FCT) through the projects B-FABULUS (PTDC/BBB-ECT/2690/2014) and Fun4TE (PTDC/EMD-EMD/31367/2017). FCT/MCTES is also acknowledged for the PhD scholarship attributed to J.B.C. (PD/BD/113803/2015) and the financial support provided to J.S.-C. (IF/00115/2015) and J.M.O. (IF/01285/2015) under the program “Investigador FCT.” The authors would like to also acknowledge the contribution of Teresa Oliveira for histology samples processing.

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Authors and Affiliations

  1. 3B´s Research Group, I3Bs – Research Institute on Biomaterials, Biodegradables and Biomimetics, University of Minho, Headquarters of the European Institute of Excellence on Tissue Engineering and Regenerative Medicine, AvePark, Zona Industrial da Gandra, 4805-017, Barco Gmr, Portugal

    João B. Costa, Joana Silva-Correia, Sandra Pina, Alain da Silva Morais, Sílvia Vieira, Hélder Pereira, João Espregueira-Mendes, Rui L. Reis & Joaquim M. Oliveira

  2. ICVS/3B’s - PT Government Associate Laboratory, Braga/Guimarães, Portugal

    João B. Costa, Joana Silva-Correia, Sandra Pina, Alain da Silva Morais, Sílvia Vieira, Hélder Pereira, João Espregueira-Mendes, Rui L. Reis & Joaquim M. Oliveira

  3. The Discoveries Centre for Regenerative and Precision Medicine, Headquarters at University of Minho, Avepark, 4805-017, Barco, Guimarães, Portugal

    João B. Costa, Rui L. Reis & Joaquim M. Oliveira

  4. Ripoll y De Prado Sports Clinic: Murcia-Madrid FIFA Medical Centre of Excellence, Murcia, Spain

    Hélder Pereira

  5. Orthopedic Department Centro Hospitalar Póvoa de Varzim, Vila do Conde, Portugal

    Hélder Pereira

  6. Clínica do Dragão, Espregueira-Mendes Sports Centre, FIFA Medical Centre of Excellence, Porto, Portugal

    João Espregueira-Mendes

  7. Dom Henrique Research Centre, Porto, Portugal

    João Espregueira-Mendes

  8. Orthopedic Department, University of Minho, Braga, Portugal

    João Espregueira-Mendes

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  1. João B. Costa
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Costa, J.B., Silva-Correia, J., Pina, S. et al. Indirect printing of hierarchical patient-specific scaffolds for meniscus tissue engineering. Bio-des. Manuf. 2, 225–241 (2019). https://doi.org/10.1007/s42242-019-00050-x

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