title: Chronic venous disease - Part II: Proteolytic biomarkers in wound healing.
abstract: Venous leg ulcers (VLU) are characterized by sustained proteolytic microenvironment impairing the healing process. Wound fluid (WF) reflect the biomolecular activities occurring within the wound area; however, it is unclear if WF from different healing phases have different proteolytic profiles and how VLU microenvironment affects the wound healing mechanisms. We investigated the proteolytic network of WF from distinct VLU phases, and in WF- and LPS-stimulated THP-1 monocytes treated with glycosaminoglycan sulodexide, a well known therapeutic approach for VLU healing. WF were collected from patients with VLU during inflammatory (Infl) and granulating (Gran) phases. WF and THP-1 supernatants were analyzed for nine matrix metalloproteinases (MMP) and four tissue inhibitors of metalloproteinases (TIMP) by multiplex immunoassays. Our results demonstrated that: 1) WF from Infl VLU contained significantly increased concentrations of MMP-2, MMP-9, MMP-12, TIMP-1, and TIMP-2 compared to Gran WF; 2) WF from Gran VLU showed significantly increased levels of MMP-1, MMP-7, MMP-13, and TIMP-4 compared to Infl WF; 3) LPS- and WF-stimulation of THP-1 cells significantly increased the expression of several MMP compared to untreated cells; 4) Sulodexide treatment of both LPS- and WF-stimulated THP-1 significantly down-regulated the release of several MMPs. Our study provides evidence-based medicine during treatment of patients with VLU. WF from Infl and Gran VLU have different MMP and TIMP signatures, consistent with their clinical state. The modulation of proteolytic pathways in wound microenvironment by glycosaminoglycan sulodexide, provide insights for translating research into clinical practice during VLU therapy.
authors.affiliation: Department of Biomolecular Sciences, Section of Clinical Biochemistry and Molecular Genetics, University "Carlo Bo", Urbino, Italy.
authors.forename: Daniela
authors.initials: D
authors.lastname: Ligi
authors.affiliation: Department of Angiology, Barbantini's Clinic, via del Calcio 2, 55100 Lucca, Italy.
authors.forename: Giovanni
authors.initials: G
authors.lastname: Mosti
authors.affiliation: Department of Biomolecular Sciences, Section of Clinical Biochemistry and Molecular Genetics, University "Carlo Bo", Urbino, Italy.
authors.forename: Lidia
authors.initials: L
authors.lastname: Croce
authors.affiliation: Vascular Surgery Division, VA Boston Healthcare System, West Roxbury, MA, USA; Harvard Medical School, Brigham and Women's Hospital, Boston, MA, USA.
authors.forename: Joseph D
authors.initials: JD
authors.lastname: Raffetto
authors.affiliation: Department of Biomolecular Sciences, Section of Clinical Biochemistry and Molecular Genetics, University "Carlo Bo", Urbino, Italy. Electronic address: ferdinando.mannello@uniurb.it.
authors.forename: Ferdinando
authors.initials: F
authors.lastname: Mannello
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D015415:Biomarkers / Q000378:metabolism; D060833:Cellular Microenvironment; D017364:Collagenases / Q000378:metabolism*; D005260:Female; D006801:Humans; D008297:Male; D008875:Middle Aged; D059748:Proteolysis; D000074084:THP-1 Cells; D019714:Tissue Inhibitor of Metalloproteinases / Q000378:metabolism*; D014647:Varicose Ulcer / Q000378:metabolism* / Q000473:pathology; D014945:Wound Healing
chemical_list: D015415:Biomarkers; D019714:Tissue Inhibitor of Metalloproteinases; D017364:Collagenases
keywords: Chronic Venous Insufficiency; Glycosaminoglycan; Matrix metalloproteinase; Tissue inhibitors of metalloproteinase; Venous leg ulcer; Wound healing