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Cyclo(RGDfK) Functionalized Spider Silk Cell Scaffolds: Significantly Improved Performance in Just One Click
Author(s) -
Harvey David,
Bray Gemma,
Zamberlan Francesco,
Amer Mahetab,
Goodacre Sara L.,
Thomas Neil R.
Publication year - 2020
Publication title -
macromolecular bioscience
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.924
H-Index - 105
eISSN - 1616-5195
pISSN - 1616-5187
DOI - 10.1002/mabi.202000255
Subject(s) - spider silk , biomaterial , silk , tissue engineering , azide , click chemistry , chemistry , biocompatibility , cell adhesion , adhesion , regenerative medicine , nanotechnology , biophysics , biochemistry , biomedical engineering , materials science , combinatorial chemistry , biology , cell , organic chemistry , medicine , composite material
Recombinant spider silk has the potential to provide a new generation of biomaterial scaffolds as a result of its degree of biocompatibility and lack of immunogenicity. These recombinant biomaterials are, however, reported to exhibit poor cellular adhesion which limits their potential for use in applications such as tissue engineering and regenerative medicine. In this study, a simple chemical functionalization approach is described that specifically addresses this issue and significantly improves the adhesion of human mesenchymal stem cells (CiMSCs) to a recombinant spider silk biomaterial. This utilizes copper‐catalyzed or strain‐promoted azide–alkyne cycloaddition (CuAAC/SPAAC) “click” chemistry to covalently attach cyclo(RGDfK) peptides to the azide group of l ‐azidohomoalanine, a methionine analogue previously site specifically incorporated into the primary sequence of a thioredoxin (TRX)‐tagged silk fusion protein, TRX‐4RepCT, to give TRX 3Aha ‐4RepCT 3Aha . This method is used to produce cyclo(RGDfK) functionalized films and macroscopic fibers. Over 24 h, cyclo(RGDfK) functionalized TRX 3Aha ‐4RepCT 3Aha films and 4RepCT 3Aha fibers display significantly improved performance in CiMSC culture, yielding far greater cell numbers than the controls. This approach circumvents the previously observed lack of cell adhesion, thus allowing spider silk derived biomaterials to be used where such adhesion is critical, in tissue engineering, regenerative medicine and wound healing.