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ACS Appl Mater Interfaces. 2019 Jul 17;11(28):24999-25007. doi: 10.1021/acsami.9b05531. Epub 2019 Jul 5.

Bioactive Silk Coatings Reduce the Adhesion of Staphylococcus aureus while Supporting Growth of Osteoblast-like Cells.

Author information

1
Department of Protein Science, School of Engineering Sciences in Chemistry, Biotechnology and Health , AlbaNova University Center, KTH Royal Institute of Technology , SE-106 91 Stockholm , Sweden.
2
Department of Molecular Biosciences, The Wenner-Gren Institute , Stockholm University , SE-106 91 Stockholm , Sweden.
3
RISE Research Institutes of Sweden , SE-11486 Stockholm , Sweden.
4
Department of Biomedical Engineering , University of Groningen and University Medical Center of Groningen , NL-9713AV Groningen , The Netherlands.

Abstract

Orthopedic and dental implants are associated with a substantial risk of failure due to biomaterial-associated infections and poor osseointegration. To prevent such outcomes, a coating can be applied on the implant to ideally both reduce the risk of bacterial adhesion and support establishment of osteoblasts. We present a strategy to construct dual-functional silk coatings with such properties. Silk coatings were made from a recombinant partial spider silk protein either alone (silkwt) or fused with a cell-binding motif derived from fibronectin (FN-silk). The biofilm-dispersal enzyme Dispersin B (DspB) and two peptidoglycan degrading endolysins, PlySs2 and SAL-1, were produced recombinantly. A sortase recognition tag (SrtTag) was included to allow site-specific conjugation of each enzyme onto silkwt and FN-silk coatings using an engineered variant of the transpeptidase Sortase A (SrtA*). To evaluate bacterial adhesion on the samples, Staphylococcus aureus was incubated on the coatings and subsequently subjected to live/dead staining. Fluorescence microscopy revealed a reduced number of bacteria on all silk coatings containing enzymes. Moreover, the bacteria were mobile to a higher degree, indicating a negative influence on the bacterial adhesion. The capability to support mammalian cell interactions was assessed by cultivation of the osteosarcoma cell line U-2 OS on dual-functional surfaces, prepared by conjugating the enzymes onto FN-silk coatings. U-2 OS cells could adhere to silk coatings with enzymes and showed high spreading and viability, demonstrating good cell compatibility.

KEYWORDS:

Staphylococcus aureus; antibacterial; endolysin; multifunctional coating; osseointegration; recombinant spider silk

PMID:
31241302
DOI:
10.1021/acsami.9b05531

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