Fibrin material and method for producing and using the same

Abstract

Abstract of the Disclosure This invention describes a bioerodible fibrin material which is obtained by mixing fibrinogen and thrombin reconstituted or diluted with a particular high tonic strength medium, free of calcium. Such a fibrin-based biomaterial develops a tight structure with thin fibers and small pore size suitable for use as an anti-adhesion barrier. In this invention, thrombin is no longer the variable which governs the tightness and the porosity of the fibrin material obtained, but still controls the clotting time. The mechanical behavior, high-water capacity, and releasable retention properties for therapeutic agents of this fibrin structure causes the fibrin material to be ideally suited for use as a drug delivery device, capable of delivering proteins, hormones, enzymes, antibiotics, antineoplastic agents and even cells for local and systemic treatment of human and non-human patients.

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       22.   A method for delivering fibrin hydrogel to a surface comprising the steps of: 
     providing a liquid solution of fibrinogen; 
     providing a liquid solution of thrombin; 
      providing a liquid solution of a calcium chelating agent; 
     providing a spray unit in fluid communication with the fibrinogen solution, the thrombin solution, and the chelating agent, the spray unit being capable of atomizing the fibrinogen solution,  the thrombin solution, and the chelating agent into an aerosol with at least one energy source of a liquid energy, a mechanical energy, a mechanical energy, and an electric energy; 
     spraying the fibrinogen solution onto the surface with the spray unit; 
     spraying the chelating agent onto the surface with the spray unit; 
      spraying the thrombin solution onto the surface with the spray unit; and 
     mixing the fibrinogen solution the chelating agent and the thrombin solution. 
   
   
       23.   The method of  claim 22  wherein the chelating agent is selected from the group of EGTA, EDTA, PBS, and citrate. 
   
   
       24.   The method of  claim 22  wherein the chelating agent is PBS. 
   
   
       25.   The method of  claim 22  wherein the step of spraying the fibrinogen solution onto the surface occurs simultaneously with the step of spraying the thrombin solution onto the surface. 
   
   
       26.   The method of  claim 22  wherein the step of mixing the fibrinogen solution and the thrombin solution defines a substantially homogeneous mixture prior to making contact with the surface. 
   
   
       27.   The method of  claim 22  wherein the step of spraying the fibrinogen solution onto the surface and the step of spraying the thrombin solution onto the surface occurs sequentially. 
   
   
       28.   A method for using a fibrin material to treat a patient, the method 
     comprising the steps of: 
     
       providing a first fibrin material having a pore size from about 2 µm to about 10 µm; 
     
     
       providing a second fibrin material having a pore size from about 0.1 µm to about 5 µm; and being capable of retaining 80-90% of water upon compression by a force of 1-14 psi; 
     
     
       applying the first fibrin material to a surface; and 
     
     applying the second fibrin material to the first fibrin material. 
   
   
       29.   The method of  claim 28  wherein the step of providing the first fibrin 
     material comprises the steps of: 
     
       providing thrombin solution having a concentration from about 50 IU/ml to about 300 IU/ml; 
     
     
       providing fibrinogen solution having a concentration from about 1.5 mg/ml to about 100 mg/ml; and 
     
     mixing the thrombin solution with the fibrinogen solution. 
   
   
       30.   The method of  claim 28  wherein the step of providing the second fibrin material 
     comprises the steps of: 
     
       providing thrombin solution having a concentration from about 1 IU/ml to about 300 IU/ml; 
     
     
       providing fibrinogen solution having a concentration from about 1.5 mg/ml to about 100 mg/ml; 
     
     
       providing a chelating agent as an antagonist to fibrinopeptide transamidation reactions; and 
     
     
       mixing the thrombin solution, the fibrinogen solution, and the chelating agent. 
     
   
   
       31.   The method of  claim 30  wherein the diluent is phosphate buffered saline. 
   
   
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       38.   A multiple layer fibrin material comprising: 
      a fibrin glue layer; and 
      a fibrin hydrogel layer, the fibrin hydrogel being capable of retaining from at least about 80% to about 90% of water by weight of the hydrogel when compressed by a force of about 156 G, and having relatively no cross-linking, wherein the fibrin hydrogel layer releasably retains a therapeutic agent. 
   
   
       39.   The multiple layer fibrin material of  claim 38  wherein the therapeutic agent is selected from the group consisting of antibiotics, fibrinolytic agents and biological response modifiers. 
   
   
       40.  The multiple layer fibrin material of  claim 39  wherein the therapeutic agent comprises a pharmaceutical compound. 
   
   
       41.   The multiple layer fibrin material of  claim 39  wherein the therapeutic agent comprises living cells. 
   
   
       42.   A multiple layer fibrin material for treating a patient comprising: 
     a fibrin glue layer; and 
     
       a therapeutic fibrin hydrogel layer. 
     
   
   
       43.   The multiple layer fibrin material of  claim 42  wherein the fibrin glue layer has a pore size from about 2 µm to about 10 µm. 
   
   
       44.   The multiple layer fibrin material of  claim 42  wherein the therapeutic fibrin hydrogel layer has a pore size from about 0.1 µm to about 5 µm. 
   
   
       45.   The multiple layer fibrin material of  claim 42 , wherein the therapeutic fibrin hydrogel layer has a water content of at least about 90% by weight of the therapeutic fibrin hydrogel layer and whereby the therapeutic fibrin hydrogel layer retains from about 80% to about 90% of the water upon compression by a force of about 156 G. 
   
   
       46.   The multiple layer fibrin material of  claim 45  wherein the retained water comprises a releasably retained diluent. 
   
   
       47.   The multiple layer fibrin material of  claim 46  wherein the releasably retained diluent comprises a therapeutic agent. 
   
   
       48.   The multiple layer fibrin material of  claim 47  wherein the therapeutic agent is selected from the group consisting of pharmaceutical compounds, antibiotics, fibrinolytic agents and biological response modifiers. 
   
   
       49.   The multiple layer fibrin material of  claim 48  wherein the therapeutic agent comprises a solution containing living cells. 
   
   
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       70.   A kit for forming a fibrin hydrogel comprising 
      a container of proteins comprising fibrinogen; 
      a container of thrombin; 
      a container of a diluent, wherein the diluent has the ability to chelate calcium; 
      and a 
      mixing and application apparatus. 
   
   
       71.  The kit of  claim 70 , wherein the container of proteins has a protein concentration of more than about 30 mg/ml. 
   
   
       72.  The kit of  claim 70 , wherein the container of proteins contains Factor XIII at a concentration from about 0 IU/ml to about 80 IU/ml. 
   
   
       73.  The kit of  claim 70 , wherein the container of thrombin has thrombin concentration between about 0.1 IU/ml and about 1000 IU/ml. 
   
   
       74.  The kit of  claim 70 , wherein the mixing and application apparatus is a syringe. 
   
   
       75.  The kit of  claim 70 , wherein the mixing and application apparatus is a catheter. 
   
   
       76.  The kit of  claim 70 , wherein the diluent is selected from the group consisting of: a phosphate buffer, sodium citrate solution, potassium citrate solution, EDTA, EGTA, and chloride solution. 
   
   
       77.  The kit of  claim 76 , wherein the diluent is the phosphate buffer. 
   
   
       78.  A kit for forming a fibrin hydrogel comprising: 
     a container of proteins comprising fibrinogen, wherein the protein concentration is more than about 30 mg/ml; 
     a container of thrombin, wherein the thrombin concentration is between about 0.1 IU/ml and about 1000 IU/ml: 
     a container of a phosphate buffer solution; and 
     a mixing and application apparatus, wherein the apparatus is selected from the group consisting of a syringe and a catheter. 
   
   
       79.  The kit of  claim 78 , wherein the container of proteins contains Factor XIII at a concentration from about 0 IU/ml to about 80 IU/ml. 
   
   
       80.  A fibrin hydrogel material comprising: 
     a fibrin material, wherein the hydrogel material having has a water content of at least 90% by weight of the hydrogel material and whereby the hydrogel material retains from about 80% to about 90% of the water upon compression by a force of about 156 G. 
   
   
       81.  A fibrin hydrogel material comprising: 
     a fibrin material, wherein the hydrogel material has a water content of at least 90% by weight of the hydrogel material and whereby the hydrogel material retains from about 80% to about 90% of the water upon compression by a force of about 156 G wherein the fibrin hydrogel material is produced using an essentially calcium free thrombin solution. 
   
   
       82.  A multiple layer fibrin material comprising: 
     
       a fibrin glue layer; and 
     
     a fibrin hydrogel layer, the fibrin hydrogel being capable of retaining from at least about 80% to about 90% of water by weight of the hydrogel when compressed by a force of about 156 G, and having relatively no cross-linking. 
   
   
       83.  A multiple layer fibrin material for treating a patient comprising: 
      a fibrin film layer; and 
      a fibrin hydrogel layer, the fibrin hydrogel having a water content of at least 90% by weight of the fibrin hydrogel and whereby the fibrin hydrogel retains from about 80% to about 90% of the water upon compression by a force of about 156 G. 
   
   
       84.  A multiple layer fibrin material for treating a patient comprising: 
      a fibrin film layer; 
      a therapeutic fibrin hydrogel layer, the fibrin hydrogel having a water content of at least 90% by weight of the fibrin hydrogel and whereby the fibrin hydrogel retains from about 80% to about 90% of the water upon compression by a force of about 156 G; and 
      a fibrin glue layer attaching the fibrin film layer to the fibrin hydrogel layer.