Matrix metalloproteinase mmp 8

EDTA-anticoagulated blood plasma collected on ICU admission was used for soluble mediator measurements. IL-6, IL-8, IL-10, soluble intercellular adhesion molecule-1 (ICAM-1), soluble E-selectin, and fractalkine were measured using FlexSet cytometric bead arrays (BD Biosciences, San Jose, CA) using a FACS Calibur (Becton Dickinson, Franklin Lakes, NJ, NJ, USA). Neutrophil gelatinase-associated lipocalin (NGAL), ANG-1, ANG-2, protein C, antithrombin, matrix metalloproteinase (MMP)−8 (R and D Systems, Abingdon, UK), and D-dimer (Procartaplex, eBioscience, San Diego, CA) were measured by Luminex multiplex assay using a BioPlex 200 (BioRas, Hercules, CA). CRP was determined by an immunoturbidimetric assay (Roche diagnostics). Platelet counts were determined by hemocytometry, prothrombin time (PT), and activated partial thromboplastin time (aPTT) by using a photometric method with Dade Innovin Reagent or by Dade Actin FS Activated PTT Reagent, respectively (Siemens Healthcare Diagnostics). Normal biomarker values were obtained from 27 age- and sex-matched healthy subjects, except for CRP, platelet counts, PT, and aPTT (routine laboratory reference values).

Daily (on admission and at 6 a.m. thereafter) EDTA anti-coagulated leftover plasma harvested from blood obtained for regular patient care was stored within 4 hours after blood draw at -80 °C. Measurements were done in EDTA anti-coagulated plasma. Tumor necrosis factor (TNF)-α, interleukin (IL)-6, IL-8, IL-1β, IL-10, IL-13, interferon-γ, soluble E-selectin, soluble intercellular adhesion molecule (ICAM)-1 and fractalkine were measured by FlexSet cytometric bead array (BD Biosciences, San Jose, CA, USA) using FACS Calibur (Becton Dickenson, Franklin Lakes, NJ, USA). Matrix metalloproteinase (MMP)-8, angiopoietin-1, angiopoietin-2, protein C, antithrombin (all R&D systems, Abingdon, UK) and D-dimer (Procartaplex, eBioscience, San Diego, CA, USA) were measured by Luminex multiplex assay using BioPlex 200 (BioRad, Hercules, CA, USA). C-reactive protein (CRP) was determined by immunoturbidimetric assay (Roche diagnostics), prothrombin time (PT) and activated partial thromboplastin time (aPTT) by using a photometric method with Dade Innovin Reagent or by Dade Actin FS Activated PTT Reagent, respectively (both Siemens Healthcare Diagnostics). Normal values were obtained in plasma from 27 age-matched and gender-matched healthy volunteers, included after providing written informed consent, with the exception of CRP, PT and aPTT (routine laboratory reference values).