title: The Influence of Chronic Wound Extracts on Inflammatory Cytokine and Histatin Stability.
abstract: Chronic ulcers represent a major health burden in our society. Despite many available therapies, a large number of ulcers do not heal. Protein based therapies fail in part due to proteolytic activity in the chronic wound bed. The aim of this in vitro study was to determine whether typical inflammatory cytokines and human salivary histatins remain stable when incubated with chronic wound extracts. Furthermore we determined whether a short exposure of histatins or cytokines was sufficient to exert long term effects on fibroblast migration. Stability of human recombinant cytokines IL-6 and CXCL8, and histatin variants (Hst1, Hst2, cyclic Hst1, minimal active domain of Hst1) in the presence of chronic wound extracts isolated from non-healing ulcers, was monitored by capillary zone electrophoresis. Migration-stimulating activity was assessed using a dermal fibroblast wound healing scratch assay. Histatins and cytokines stayed stable in saline for > 24 h at 37°C, making them ideal as an off-the-shelf product. However, incubation with chronic wound extracts resulted in serious breakdown of Hst1 and Hst2 (~50% in 8 h) and to lesser extent cyclic Hst1 and the minimal active domain of Hst1 (~20% in 8 h). The cytokines IL-6 and CXCL8 were more stable in chronic wound extracts (~40% degradation in 96 h). An initial 8-hour pulse of histatins or cytokines during a 96-hour study period was sufficient to stimulate fibroblast migration equally well as a continuous 96-hour exposure, indicating that they may possibly be used as novel bioactive therapeutics, exerting their activity for up to four days after a single exposure.
authors.affiliation: Department of Oral Biochemistry, Academic Centre for Dentistry Amsterdam (ACTA), University of Amsterdam and VU University Amsterdam, Amsterdam, The Netherlands.|Department of Dermatology, VU University medical center, Amsterdam, The Netherlands.
authors.forename: Mireille A
authors.initials: MA
authors.lastname: Boink
authors.affiliation: Department of Oral Biochemistry, Academic Centre for Dentistry Amsterdam (ACTA), University of Amsterdam and VU University Amsterdam, Amsterdam, The Netherlands.|Department of Dermatology, VU University medical center, Amsterdam, The Netherlands.
authors.forename: Sanne
authors.initials: S
authors.lastname: Roffel
authors.affiliation: Department of Oral Biochemistry, Academic Centre for Dentistry Amsterdam (ACTA), University of Amsterdam and VU University Amsterdam, Amsterdam, The Netherlands.
authors.forename: Kamran
authors.initials: K
authors.lastname: Nazmi
authors.affiliation: Department of Dermatology, VU University medical center, Amsterdam, The Netherlands.
authors.forename: Catherine
authors.initials: C
authors.lastname: van Montfrans
authors.affiliation: Department of Oral Biochemistry, Academic Centre for Dentistry Amsterdam (ACTA), University of Amsterdam and VU University Amsterdam, Amsterdam, The Netherlands.
authors.forename: Jan G M
authors.initials: JG
authors.lastname: Bolscher
authors.affiliation: Department of Biomedical Engineering, Faculty of Engineering, Tel Aviv University, Tel Aviv, Israel.
authors.forename: Amit
authors.initials: A
authors.lastname: Gefen
authors.affiliation: Department of Oral Biochemistry, Academic Centre for Dentistry Amsterdam (ACTA), University of Amsterdam and VU University Amsterdam, Amsterdam, The Netherlands.
authors.forename: Enno C I
authors.initials: EC
authors.lastname: Veerman
authors.affiliation: Department of Dermatology, VU University medical center, Amsterdam, The Netherlands.|Department of Oral Cell Biology, Academic Centre for Dentistry Amsterdam (ACTA), University of Amsterdam and VU University, MOVE Research Institute Amsterdam, Amsterdam, The Netherlands.
authors.forename: Susan
authors.initials: S
authors.lastname: Gibbs
mesh_terms: D002465:Cell Movement / Q000187:drug effects; D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid; D016207:Cytokines / Q000737:chemistry / Q000378:metabolism*; D019075:Electrophoresis, Capillary; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism; D054808:Histatins / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D015850:Interleukin-6 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D016209:Interleukin-8 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D007871:Leg Ulcer / Q000378:metabolism / Q000473:pathology; D055550:Protein Stability; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000302:isolation & purification / Q000494:pharmacology
chemical_list: D016207:Cytokines; D054808:Histatins; D015850:Interleukin-6; D016209:Interleukin-8; D011994:Recombinant Proteins