Biomaterials, Biodegradables and Biomimetics Research Group

Papers in Scientific Journals

Antimicrobial functionalized genetically engineered spider silk

Abstract

Genetically engineered fusion proteins offer potential as multifunctional biomaterials for medical use. Fusion or chimeric proteins can be formed using recombinant DNA technology by combining nucleotide sequences encoding different peptides or proteins that are otherwise not found together in nature. In the present study, three new fusion proteins were designed, cloned and expressed and assessed for function, by combining the consensus sequence of dragline spider silk with three different antimicrobial peptides. The human antimicrobial peptides human neutrophil defensin 2 (HNP-2), human neutrophil defensins 4 (HNP-4) and hepcidin were fused to spider silk through bioengineering. The spider silk domain maintained its self-assembly features, a key aspect of these new polymeric protein biomaterials, allowing the formation of beta-sheets to lock in structures via physical interactions without the need for chemical cross-linking. These new functional silk proteins were assessed for antimicrobial activity against Gram - Escherichia coli and Gram + Staphylococcus aureus and microbicidal activity was demonstrated. Dynamic light scattering was used to assess protein aggregation to clarify the antimicrobial patterns observed. Attenuated-total reflectance Fourier transform infrared spectroscopy (ATR-FTIR) and circular dichroism (CD) were used to assess the secondary structure of the new recombinant proteins. In vitro cell studies with a human osteosarcoma cell line (SaOs-2) demonstrated the compatibility of these new proteins with mammalian cells. (C) 2011 Elsevier Ltd. All rights reserved.
Journal
Biomaterials
Volume
32
Issue
18
Pagination
4255-4266
Publisher
Elsevier
ISSN
0142-9612
URL
http://www.sciencedirect.com/science/article/pii/S0142961211002031
Keywords
Antimicrobial activity, Bone Tissue Engineering, Cell viability, Recombinant proteins, self-assembly, Spider silk
Rights
Restricted Access
Peer Reviewed
Yes
Status
published
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