Covalent Protein Immobilization on 3D-Printed Microfiber Meshes for Guided Cartilage Regeneration
Ainsworth, Madison J.; Lotz, Oliver; Gilmour, Aaron; Zhang, Anyu; Chen, Michael J.; McKenzie, David R.; Bilek, Marcela M.M.; Malda, Jos; Akhavan, Behnam; Castilho
(2023) Advanced Functional Materials, issue 2
(Article)
Abstract
Current biomaterial-based strategies explored to treat articular cartilage defects have failed to provide adequate physico-chemical cues in order to guide functional tissue regeneration. Here, it is hypothesized that atmospheric-pressure plasma (APPJ) treatment and melt electrowriting (MEW) will produce microfiber support structures with covalently-immobilized transforming growth factor beta-1 (TGFβ1) that can
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stimulate the generation of functional cartilage tissue. The effect of APPJ operational speeds to activate MEW polycaprolactone meshes for immobilization of TGFβ1 is first investigated and chondrogenic differentiation and neo-cartilage production are assessed in vitro. All APPJ speeds test enhanced hydrophilicity of the meshes, with the slow treatment speed having significantly less C C/C H and more COOH than the untreated meshes. APPJ treatment increases TGFβ1 loading efficiency. Additionally, in vitro experiments highlight that APPJ-based TGFβ1 attachment to the scaffolds is more advantageous than direct supplementation within the medium. After 28 days of culture, the group with immobilized TGFβ1 has significantly increased compressive modulus (more than threefold) and higher glycosaminoglycan production (more than fivefold) than when TGFβ1 is supplied through the medium. These results demonstrate that APPJ activation allows reagent-free, covalent immobilization of TGFβ1 on microfiber meshes and, importantly, that the biofunctionalized meshes can stimulate neo-cartilage matrix formation. This opens new perspectives for guided tissue regeneration.
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Keywords: atmospheric-pressure plasma, cartilage, melt electrowriting, protein immobilization, stem cell differentiation, technology convergence, transforming growth factor beta, General Chemistry, General Materials Science, Condensed Matter Physics
ISSN: 1616-301X
Publisher: Wiley-VCH Verlag
Note: Funding Information: M.J.A. and O.L. contributed equally to this work. B.A. and M.C. are shared senior authors. The authors would like to kindly acknowledge the financial support from the Gravitation Program “Materials Driven Regeneration”, funded by the Netherlands Organization for Scientific Research (024.003.013), the EU's H2020 Marie Skłodowska-Curie RESCUE co-fund grant (#801540), the Jennifer Foong Scholarship for Biomedical Research, and an Office of Global Engagement Partnership Collaboration Award between the University of Sydney, Utrecht University, and the Australian Research Council (FL190100216; DP190103507; and DE210100662). This work was also supported by the partners of Regenerative Medicine Crossing Borders and powered by Health ∼Holland, Top Sector Life Sciences & Health. The authors would like to thank I. Dokter for assistance with running qPCRs and M. van Rijen for histology expertise. The DSHB Hybridoma Product II-II6B3 developed by T.F. Linsenmayer was obtained from the Developmental Studies Hybridoma Bank, created by the NICHD of the NIH and maintained at The University of Iowa, Department of Biology, Iowa City, IA 52242. Funding Information: M.J.A. and O.L. contributed equally to this work. B.A. and M.C. are shared senior authors. The authors would like to kindly acknowledge the financial support from the Gravitation Program “Materials Driven Regeneration”, funded by the Netherlands Organization for Scientific Research (024.003.013), the EU's H2020 Marie Skłodowska‐Curie RESCUE co‐fund grant (#801540), the Jennifer Foong Scholarship for Biomedical Research, and an Office of Global Engagement Partnership Collaboration Award between the University of Sydney, Utrecht University, and the Australian Research Council (FL190100216; DP190103507; and DE210100662). This work was also supported by the partners of Regenerative Medicine Crossing Borders and powered by Health ∼Holland, Top Sector Life Sciences & Health. The authors would like to thank I. Dokter for assistance with running qPCRs and M. van Rijen for histology expertise. The DSHB Hybridoma Product II‐II6B3 developed by T.F. Linsenmayer was obtained from the Developmental Studies Hybridoma Bank, created by the NICHD of the NIH and maintained at The University of Iowa, Department of Biology, Iowa City, IA 52242. Publisher Copyright: © 2022 The Authors. Advanced Functional Materials published by Wiley-VCH GmbH.
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