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Enzyme-Mediated Conjugation of Peptides to Silk Fibroin for Facile Hydrogel Functionalization

  • Biomaterials - Engineering Cell Behavior
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Abstract

Enzymatic crosslinking of tyrosine is a simple and modular method for adding functional peptides to silk fibroin (SF) hydrogels. Silk fibroin is a naturally derived polymer notable for its robust mechanical properties, biological compatibility, and versatility. Hydrogels fabricated from SF are elastic, optically clear, and have tunable moduli, however, they do not contain native biological epitopes for cell interactions. In this work we demonstrate the attachment of peptides to SF hydrogels through crosslinking of tyrosine with horseradish peroxidase (HRP) and hydrogen peroxide (H2O2). The goal was to understand the utility of this approach and to study how the addition of peptides affects the SF material properties. SF hydrogels conjugated to model peptides with different molecular weights and hydrophobic properties were studied by liquid chromatography/tandem mass spectroscopy (LC–MS/MS) (bond formation), fluorescent imaging (spatial distribution), Fourier transform infrared spectroscopy (FTIR) (protein secondary structure), and rheology (gelation time, modulus). As a proof of concept using a biologically relevant peptide, a peptide containing the cell binding domain Arg-Gly-Asp (RGD) was conjugated to SF, and the density and morphology of primary human fibroblasts were assessed. This work demonstrates a facile method for adding peptides to silk fibroin that can be adopted for a variety of biomaterials applications.

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Acknowledgments

This work was supported by the Air Force Office of Scientific Research under Grant No. FA9550-17-1-0333, and the National Science Foundation Graduate Research Fellowship under Grant No. DGE-1325256 and the NIH (P41EB027062). We thank Jim Harden, U. Ottawa for helpful suggestions and discussion and Jon Grasman for helping to plan the in vitro experiments.

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  1. Department of Biomedical Engineering, Tufts University, 4 Colby Street, Medford, MA, 02155, USA

    Meghan McGill, James M. Grant & David L. Kaplan

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  1. Meghan McGill
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Correspondence to David L. Kaplan.

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McGill, M., Grant, J.M. & Kaplan, D.L. Enzyme-Mediated Conjugation of Peptides to Silk Fibroin for Facile Hydrogel Functionalization. Ann Biomed Eng 48, 1905–1915 (2020). https://doi.org/10.1007/s10439-020-02503-2

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  • DOI: https://doi.org/10.1007/s10439-020-02503-2

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