Detection of human neutrophil elastase (HNE) on wound dressings as marker of inflammation

Ana V. Ferreira, Ilana Perelshtein, Nina Perkas, Aharon Gedanken, Joana Cunha, Artur Cavaco-Paulo

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25 Scopus citations

Abstract

Chronic wound fluids have elevated concentration of human neutrophil elastase (HNE) which can be used as inflammation/infection marker. Our goal is to develop functional materials for fast diagnosis of wound inflammation/infection by using HNE as a specific marker. For that, fluorogenic peptides with a HNE-specific cleavage sequence were incorporated into traditional textile dressings, to allow real-time detection of the wound status. Two different fluorogenic approaches were studied in terms of intensity of the signal generated upon HNE addition: a fluorophore 7-amino-4-trifluormethylcoumarin (AFC) conjugated to a HNE-specific peptide and two fluorophore/quencher pairs (FAM/Dabcyl and EDANS/Dabcyl) coupled to a similar peptide as a Förster resonance energy transfer (FRET) strategy. Also, two immobilization methods were tested: sonochemistry immobilization onto a cotton bandage and glutaraldehyde (GTA)-assisted chemical crosslinking onto a polyamide dressing. The immobilized fluorogenic AFC peptide showed an intense fluorescence emission in the presence of HNE. HNE also induced an enhanced fluorescent signal with the EDANS/Dabcyl FRET peptide which showed to be a more sensitive and effective strategy than the AFC peptide. However, its chemical immobilization onto the polyamide dressing greatly decreased its detection, mainly due to the more difficult access of the enzyme to the cleavage sequence of the immobilized peptide. After optimization of the in situ immobilization, it will be possible to use these fluorescence-functionalized dressings for an effective and specific monitoring of chronic wounds by simply using a portable ultraviolet (UV) light source. We envision that the development of this point-of-care medical device for wound control will have a great impact on patient’s life quality and reduction of costs on health care system.

Original languageEnglish
Pages (from-to)1443-1454
Number of pages12
JournalApplied Microbiology and Biotechnology
Volume101
Issue number4
DOIs
StatePublished - 1 Feb 2017

Bibliographical note

Publisher Copyright:
© 2016, Springer-Verlag Berlin Heidelberg.

Funding

We would like to acknowledge the European project InFact—Functional materials for fast diagnosis of wound infection (FP7-NMP-2013-SME-7—grant agreement no. 604278) for funding the work and also the grant for Joana Cunha. Ana Ferreira would also like to acknowledge the Portuguese Foundation for Science and Technology (FCT) for the PhD student grant SFRH/BD/113247/2015. The work done at Centre of Biological Engineering (CEB) was supported by FCT under the scope of the strategic funding of UID/BIO/04469/2013 unit, COMPETE 2020 (POCI-01-0145-FEDER-006684) and BioTecNorte operation (NORTE-01-0145-FEDER-000004) funded by European Regional Development Fund under the scope of Norte 2020—Programa Operacional Regional do Norte. This study was funded by the European project InFact—Functional materials for fast diagnosis of wound infection (FP7-NMP-2013-SME-7—grant agreement no. 604278). The work done at Centre of Biological Engineering (CEB) was also supported by the Portuguese Foundation for Science and Technology (FCT) under the scope of the strategic funding of UID/BIO/04469/2013 unit, COMPETE 2020 (POCI-01-0145-FEDER-006684) and BioTecNorte operation (NORTE-01-0145-FEDER-000004) funded by European Regional Development Fund under the scope of Norte 2020—Programa Operacional Regional do Norte.

FundersFunder number
FP7-NMP-2013-SME-7
Seventh Framework Programme604278
Fundação para a Ciência e a TecnologiaNORTE-01-0145-FEDER-000004, SFRH/BD/113247/2015, UID/BIO/04469/2013, POCI-01-0145-FEDER-006684
European Regional Development Fund

    Keywords

    • Chronic wounds
    • Fluorogenic detection
    • Functionalized wound dressing
    • Förster resonance energy transfer (FRET)
    • Human neutrophil elastase (HNE)

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