The development of photochemically crosslinked native fibrinogen as a rapidly formed and mechanically strong surgical tissue sealant.

We recently reported the generation of a highly elastic, crosslinked protein biomaterial via a rapid photochemical process using visible light illumination. In light of these findings, we predicted that other unmodified, tyrosine-rich, self-associating proteins might also be susceptible to this covalent crosslinking method. Here we show that unmodified native fibrinogen can also be photochemically crosslinked into an elastic hydrogel biomaterial through the rapid formation of intermolecular dityrosine. Photochemically crosslinked fibrinogen forms tissue sealant bonds at least 5-fold stronger than commercial fibrin glue and is capable of producing maximum bond strength within 20s. In vitro studies showed that components of the photochemical crosslinking reaction are non-toxic to cells. This material will find useful application in various surgical procedures where rapid curing for high strength tissue sealing is required.

[1]  R. Karpel,et al.  Free radical-induced fibrinogen coagulation: modulation of neofibe formation by concentration, pH and temperature. , 1991, Israel journal of medical sciences.

[2]  D. Browdie,et al.  Tests of experimental tissue adhesive sealants: analysis of strength effects in relation to tissue adhesive sealant standards. , 2007, Texas Heart Institute journal.

[3]  R. Saltz,et al.  Mechanical properties of fibrin adhesives for blood vessel anastomosis. , 1992, Journal of biomedical materials research.

[4]  M. Huggins Viscoelastic Properties of Polymers. , 1961 .

[5]  P. Messersmith,et al.  Enzymatically cross-linked hydrogels and their adhesive strength to biosurfaces. , 2005, Orthodontics & craniofacial research.

[6]  O. Malka,et al.  The binding of fibrin sealant to collagen is influenced by the method of purification and the cross-linked fibrinogen–fibronectin (heteronectin) content of the ‘fibrinogen’ component , 2005, Blood coagulation & fibrinolysis : an international journal in haemostasis and thrombosis.

[7]  K. Siebenlist,et al.  The Structure and Biological Features of Fibrinogen and Fibrin , 2001, Annals of the New York Academy of Sciences.

[8]  Meng-G Martin Lee,et al.  Applications of Fibrin Sealant in Surgery , 2005, Surgical innovation.

[9]  T. Kodadek,et al.  Chemistry for the analysis of protein-protein interactions: rapid and efficient cross-linking triggered by long wavelength light. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[10]  H. Kjaergård,et al.  Comparative kinetics of polymerisation of three fibrin sealants and influence on timing of tissue adhesion. , 2000, Thrombosis research.

[11]  J. Vuichoud,et al.  Quantitative determination of dityrosine in milk powders by liquid chromatography coupled to tandem mass spectrometry using isotope dilution. , 2004, Journal of chromatography. A.

[12]  Steven M. Alston,et al.  New method to prepare autologous fibrin glue on demand. , 2007, Translational research : the journal of laboratory and clinical medicine.

[13]  T. Pulawska,et al.  Development of a Model for Measurement of Adhesion Strength of Fibrin Sealant to Human Tissue , 1999, European Surgical Research.

[14]  T. Vuocolo,et al.  Synthesis and properties of crosslinked recombinant pro-resilin , 2005, Nature.

[15]  S. Bergel Ueber Wirkungen des Fibrins , 1909 .

[16]  M. K. McDermott,et al.  Mechanical properties of biomimetic tissue adhesive based on the microbial transglutaminase-catalyzed crosslinking of gelatin. , 2004, Biomacromolecules.

[17]  L. Medved,et al.  Interaction of the fibronectin COOH-terminal Fib-2 regions with fibrin: further characterization and localization of the Fib-2-binding sites. , 2007, Biochemistry.

[18]  Alan W Eberhardt,et al.  Failure characteristics of multiple-component fibrin-based adhesives. , 2002, Journal of biomedical materials research.

[19]  T. Orr,et al.  Attachment of periosteal grafts to articular cartilage with fibrin sealant. , 1999, Journal of biomedical materials research.

[20]  G. Rodeheaver,et al.  Modulation of mechanical properties in multiple-component tissue adhesives. , 2000, Journal of biomedical materials research.

[21]  T. Uchida,et al.  Increased effectiveness of fibrin sealant with a higher fibrin concentration , 2000 .

[22]  C. Buckley,et al.  Bond strength of fibrin glue between layers of porcine small intestine submucosa (SIS). , 2002, Biomedical sciences instrumentation.

[23]  T. Kodadek,et al.  Techniques: Oxidative cross-linking as an emergent tool for the analysis of receptor-mediated signalling events. , 2005, Trends in pharmacological sciences.

[24]  Y. Kato,et al.  Effects of Peroxidase and Hydrogen Peroxide on the Dityrosine Formation and the Mixing Characteristics of Wheat-Flour Dough , 2005, Bioscience, biotechnology, and biochemistry.

[25]  T. Podor,et al.  Incorporation of Vitronectin into Fibrin Clots , 2002, The Journal of Biological Chemistry.

[26]  S. Brew,et al.  Cryptic Self-association Sites in Type III Modules of Fibronectin* , 1997, The Journal of Biological Chemistry.

[27]  Todd A Blackledge,et al.  Silken toolkits: biomechanics of silk fibers spun by the orb web spider Argiope argentata (Fabricius 1775) , 2006, Journal of Experimental Biology.

[28]  K. Kar,et al.  Self-association of Collagen Triple Helic Peptides into Higher Order Structures* , 2006, Journal of Biological Chemistry.

[29]  H. Hörmann,et al.  Interaction of fibrinogen/fibrin and fibronectin with collagen. , 1980, Artery.

[30]  G. Marx,et al.  Characterizing fibrin glue performance as modulated by heparin, aprotinin, and factor XIII. , 2002, The Journal of laboratory and clinical medicine.

[31]  R. Carbon,et al.  Evaluating the in vitro adhesive strength of biomaterials. Biosimulator for selective leak closure. , 2003, Biomaterials.

[32]  D. Malencik,et al.  Dityrosine: preparation, isolation, and analysis. , 1996, Analytical biochemistry.

[33]  S. Brew,et al.  Type I collagen contains at least 14 cryptic fibronectin binding sites of similar affinity. , 2002, Archives of biochemistry and biophysics.

[34]  Misook Kim,et al.  A synthetic resilin is largely unstructured. , 2008, Biophysical journal.