title: Detection of human neutrophil elastase (HNE) on wound dressings as marker of inflammation.
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.
authors.affiliation: Centre of Biological Engineering (CEB), University of Minho, 4710-057, Braga, Portugal.
authors.forename: Ana V
authors.initials: AV
authors.lastname: Ferreira
authors.affiliation: Department of Chemistry, Bar-Ilan Institute of Nanotechnology and Advanced Materials, Bar-Ilan University, IL-52900, Ramat-Gan, Israel.
authors.forename: Ilana
authors.initials: I
authors.lastname: Perelshtein
authors.affiliation: Department of Chemistry, Bar-Ilan Institute of Nanotechnology and Advanced Materials, Bar-Ilan University, IL-52900, Ramat-Gan, Israel.
authors.forename: Nina
authors.initials: N
authors.lastname: Perkas
authors.affiliation: Department of Chemistry, Bar-Ilan Institute of Nanotechnology and Advanced Materials, Bar-Ilan University, IL-52900, Ramat-Gan, Israel.
authors.forename: Aharon
authors.initials: A
authors.lastname: Gedanken
authors.affiliation: Centre of Biological Engineering (CEB), University of Minho, 4710-057, Braga, Portugal.
authors.forename: Joana
authors.initials: J
authors.lastname: Cunha
authors.affiliation: Centre of Biological Engineering (CEB), University of Minho, 4710-057, Braga, Portugal. artur@deb.uminho.pt.
authors.forename: Artur
authors.identifier: http://orcid.org/0000-0001-7204-2064
authors.initials: A
authors.lastname: Cavaco-Paulo
mesh_terms: D001458:Bandages; D031541:Fluorescence Resonance Energy Transfer; D006801:Humans; D007249:Inflammation / Q000201:enzymology*; D019272:Leukocyte Elastase / Q000378:metabolism*; D014466:Ultraviolet Rays
chemical_list: D019272:Leukocyte Elastase
keywords: Chronic wounds; Fluorogenic detection; Functionalized wound dressing; Förster resonance energy transfer (FRET); Human neutrophil elastase (HNE)