The fibrinogenolytic activity was evaluated by incubating 20 [micro]g of human fibrinogen
(Sigma Aldrich) with 5 [micro]g of the crude extract for 15, 30, 45, 60 and 120 min.
It consists of an equine collagen patch coated with fibrin glue components: human fibrinogen
and human thrombin.
Gelrin is a biodegradable hydrogel based on polyethylene glycol diacrylate and denatured human fibrinogen
originally developed at the Technion - Israel Institute of Technology.
is a soluble, 46 nm long, 340 kDa glycoprotein and is the third most prevalent protein found in blood plasma, circulating at 6-12 [micro]M .
and Rat type IV Collagen were purchased from Sigma Chemical (St.
The reagents used in this study are fibrinogen fluorescence dye (Alexa Fluor 488 human fibrinogen
conjugates (F-13191) purchased from Molecular Probes and prepared as per manufacturer's instructions.
Lyophilized human fibrinogen
(Sigma-Aldrich, Prague, Czech Republic) (4mg/mL) was dissolved in phosphate buffer saline (PBS; 137 M NaCl, 2.7 mM KCl, 8 mM [Na.sub.2]HP[O.sub.4] x 12[H.sub.2]O,1.5 mM K[H.sub.2]P[O.sub.4], and pH 7.4).
(19.) Dhall TZ, Bryce WA, Dhall DP: Effects of dextran on the molecular structure and tensile behavior of human fibrinogen
. Thromb Haemost 1976; 35:737-745.
The different protein coating solutions consisted of 2 mg/ml human albumin (Pharmacia & Upjohn AB, Stockholm, Sweden), 2 mg/ml human fibrinogen
(American Diagnostica Inc., Greenwich, Connecticut, USA), 10 pg/ml human fibronectin (Roche Diagnostics, Mannheim, Germany) or 0.1 mg/ml collagen S from calf skin (Roche Diagnostics) combined with 2 pg/ml horse tendon collagen (Biopool International, Ventura, California, USA).
Plasminogen-free unfractionated human fibrinogen
was obtained from Calbiochem.