A mechanistic investigation of the effect of keratin-based hemostatic agents on coagulation.

Uncontrolled bleeding continues to be one of the leading causes of death in individuals following traumatic injury. Prognosis is worsened with the onset of acute coagulopathy characterized by metabolic acidosis, hypothermia and hemodilution, which consequently perpetuates blood loss and increases mortality. While there are several limitations to biomaterials employed as hemostatic agents, keratin biomaterials have demonstrated efficacy in mitigating blood loss in an animal model of hemorrhage in prior studies. Here we investigate the hypothesis that keratins actively participate in coagulation and that a potential mechanism of action is independent of temperature and dilution of clotting factors. Data from this study show that keratins appear to contribute to hemostasis by significantly decreasing plasma clotting lag times and are able to maintain activity under simulated conditions of coagulopathy. Moreover, a system of isolated fibrin polymerization provided evidence of increased fibril lateral assembly in the presence of keratin. The data provided here provides a platform for further development of keratin biomaterials as hemostatic agents.

[1]  J. Jui,et al.  Multicenter hypothermia survey. , 1987, Annals of emergency medicine.

[2]  F. Bongard,et al.  Hypothermia in trauma patients. , 1999, Journal of the American College of Surgeons.

[3]  Lisa L. Schlitzkus,et al.  Impact of hypothermia (below 36 degrees C) in the rural trauma patient. , 2009, Journal of the American College of Surgeons.

[4]  B. Seifert,et al.  Low- and Medium-molecular-weight Hydroxyethyl Starches: Comparison of Their Effect on Blood Coagulation , 2000, Anesthesiology.

[5]  E. Bulger,et al.  Death in the operating room : an analysis of a multi-center experience , 1994 .

[6]  W. Chandler,et al.  Comparing the prothrombin time INR versus the APTT to evaluate the coagulopathy of acute trauma. , 2007, Thrombosis research.

[7]  J. Hermans,et al.  Size and density of fibrin fibers from turbidity. , 1978, Macromolecules.

[8]  L. Lorand,et al.  Structural origins of fibrin clot rheology. , 1999, Biophysical journal.

[9]  Michael A Dubick,et al.  Damage control resuscitation: directly addressing the early coagulopathy of trauma. , 2007, The Journal of trauma.

[10]  R. Colman,et al.  Hemostasis and Thrombosis: Basic Principles and Clinical Practice , 1988 .

[11]  A. Hirshberg,et al.  Minimizing dilutional coagulopathy in exsanguinating hemorrhage: a computer simulation. , 2003, The Journal of trauma.

[12]  F. Stuber,et al.  Preconditions of hemostasis in trauma: a review. The influence of acidosis, hypocalcemia, anemia, and hypothermia on functional hemostasis in trauma. , 2008, The Journal of trauma.

[13]  Kenichi A. Tanaka,et al.  Prothrombin complex concentrate versus recombinant factor VIIa for reversal of hemodilutional coagulopathy in a porcine trauma model. , 2010, The Journal of trauma.

[14]  E. Moore,et al.  Thomas G. Orr Memorial Lecture. Staged laparotomy for the hypothermia, acidosis, and coagulopathy syndrome. , 1996, American journal of surgery.

[15]  M. V. Van Dyke,et al.  Vasoactive Properties of Keratin‐Derived Compounds , 2011, Microcirculation.

[16]  J. Holcomb,et al.  Coagulopathy: Its Pathophysiology and Treatment in the Injured Patient , 2007, World Journal of Surgery.

[17]  Kenichi A. Tanaka,et al.  Finding the optimal concentration range for fibrinogen replacement after severe haemodilution: an in vitro model. , 2009, British journal of anaesthesia.

[18]  L. Gentilello,et al.  Continuous arteriovenous rewarming: report of a new technique for treating hypothermia. , 1991, The Journal of trauma.

[19]  A. Ray,et al.  Preparation of scaffolds from human hair proteins for tissue-engineering applications , 2008, Biomedical materials.

[20]  I. H. Segel Enzyme Kinetics: Behavior and Analysis of Rapid Equilibrium and Steady-State Enzyme Systems , 1975 .

[21]  P. Neame,et al.  Coagulopathy related to dilution and hypotension during massive transfusion , 1985, Critical care medicine.

[22]  E. Bennett-Guerrero,et al.  A Head-to-Head Comparison of the In Vitro Coagulation Effects of Saline-Based and Balanced Electrolyte Crystalloid and Colloid Intravenous Fluids , 2006, Anesthesia and analgesia.

[23]  R. Millner,et al.  A review of topical hemostatic agents for use in cardiac surgery. , 2009, The Annals of thoracic surgery.

[24]  G. Jurkovich,et al.  Hypothermia in Trauma Victims: An Ominous Predictor of Survival , 1988 .

[25]  G. Isbister,et al.  A turbidimetric assay for the measurement of clotting times of procoagulant venoms in plasma. , 2010, Journal of pharmacological and toxicological methods.

[26]  A. Wolberg,et al.  Analyzing fibrin clot structure using a microplate reader , 2002, Blood coagulation & fibrinolysis : an international journal in haemostasis and thrombosis.

[27]  R. Fischer,et al.  The disparity between hypothermic coagulopathy and clotting studies. , 1992, The Journal of trauma.

[28]  Kenichi A. Tanaka,et al.  Haemodilution-induced profibrinolytic state is mitigated by fresh-frozen plasma: implications for early haemostatic intervention in massive haemorrhage. , 2010, British journal of anaesthesia.

[29]  Kenichi A. Tanaka,et al.  Pathophysiology and Treatment of Coagulopathy in Massive Hemorrhage and Hemodilution , 2010, Anesthesiology.

[30]  J. Shafer,et al.  A kinetic model for the alpha-thrombin-catalyzed conversion of plasma levels of fibrinogen to fibrin in the presence of antithrombin III. , 1991, The Journal of biological chemistry.

[31]  Anthony Atala,et al.  A keratin biomaterial gel hemostat derived from human hair: evaluation in a rabbit model of lethal liver injury. , 2008, Journal of biomedical materials research. Part B, Applied biomaterials.

[32]  P. Kilgo,et al.  INJURY-ASSOCIATED HYPOTHERMIA: AN ANALYSIS OF THE 2004 NATIONAL TRAUMA DATA BANK , 2005, Shock.

[33]  Bantayehu Sileshi,et al.  A Comprehensive Review of Topical Hemostatic Agents: Efficacy and Recommendations for Use , 2010, Annals of surgery.

[34]  G. Gravlee Antithrombin Deficiency Increases Thrombin Activity After Prolonged Cardiopulmonary Bypass , 2009 .

[35]  D. Mosher,et al.  Effects of thrombospondin on fibrin polymerization and structure. , 1986, The Journal of biological chemistry.

[36]  M. V. Van Dyke,et al.  Mechanisms of hepatocyte attachment to keratin biomaterials. , 2011, Biomaterials.

[37]  T. Tanabe,et al.  Fabrication of wool keratin sponge scaffolds for long-term cell cultivation. , 2002, Journal of biotechnology.

[38]  K. Carlsson,et al.  Fibrin in human plasma: gel architectures governed by rate and nature of fibrinogen activation. , 1994, Thrombosis research.

[39]  M. Carr,et al.  Dextran-induced Changes in Fibrin Fiber Size and Density Based on Wavelength Dependence of Gel Turbidity , 1980 .

[40]  Alisa S Wolberg,et al.  Thrombin generation and fibrin clot structure. , 2007, Blood reviews.

[41]  R. Lefering,et al.  Early coagulopathy in multiple injury: an analysis from the German Trauma Registry on 8724 patients. , 2007, Injury.

[42]  M. Loudon,et al.  Damage control resuscitation for patients with major trauma , 2009, BMJ : British Medical Journal.

[43]  B. Blombäck,et al.  Calcium and fibrin gel structure. , 1983, Thrombosis research.

[44]  R. Suojaranta-Ylinen,et al.  Gelatin and Hydroxyethyl Starch, but Not Albumin, Impair Hemostasis After Cardiac Surgery , 2006, Anesthesia and analgesia.

[45]  A. Minton,et al.  Acceleration of fibrin gel formation by unrelated proteins. , 1985, Thrombosis research.

[46]  Karim Brohi,et al.  Functional definition and characterization of acute traumatic coagulopathy , 2011, Critical care medicine.

[47]  M. J. Rohrer,et al.  Effect of hypothermia on the coagulation cascade , 1992, Critical care medicine.

[48]  Mauricio Lynn,et al.  Early coagulopathy predicts mortality in trauma. , 2003, The Journal of trauma.

[49]  M. Carr,et al.  The effect of dextran 70 on the structure of plasma-derived fibrin gels. , 1980, The Journal of laboratory and clinical medicine.

[50]  A. Wolberg,et al.  Elevated prothrombin results in clots with an altered fiber structure: a possible mechanism of the increased thrombotic risk. , 2003, Blood.

[51]  J. Hermans,et al.  Assembly of fibrin. A light scattering study. , 1979, The Journal of biological chemistry.