US20060155234A1 - Hemostatic dressing - Google Patents

Hemostatic dressing Download PDF

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Publication number
US20060155234A1
US20060155234A1 US10/525,415 US52541506A US2006155234A1 US 20060155234 A1 US20060155234 A1 US 20060155234A1 US 52541506 A US52541506 A US 52541506A US 2006155234 A1 US2006155234 A1 US 2006155234A1
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United States
Prior art keywords
layer
fibrinogen
thrombin
hemostatic dressing
dressing
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Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Abandoned
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US10/525,415
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English (en)
Inventor
Martin Macphee
Dawson Beall
Stanley Friedman
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American National Red Cross
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American National Red Cross
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Publication date
Application filed by American National Red Cross filed Critical American National Red Cross
Priority to US10/525,415 priority Critical patent/US20060155234A1/en
Assigned to AMERICAN NATIONAL RED CROSS reassignment AMERICAN NATIONAL RED CROSS ASSIGNMENT OF ASSIGNORS INTEREST (SEE DOCUMENT FOR DETAILS). Assignors: MACPHEE, MARTIN, FRIEDMAN, STANLEY, BEALL, DAWSON
Publication of US20060155234A1 publication Critical patent/US20060155234A1/en
Priority to US13/630,890 priority patent/US8679528B2/en
Abandoned legal-status Critical Current

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Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61F—FILTERS IMPLANTABLE INTO BLOOD VESSELS; PROSTHESES; DEVICES PROVIDING PATENCY TO, OR PREVENTING COLLAPSING OF, TUBULAR STRUCTURES OF THE BODY, e.g. STENTS; ORTHOPAEDIC, NURSING OR CONTRACEPTIVE DEVICES; FOMENTATION; TREATMENT OR PROTECTION OF EYES OR EARS; BANDAGES, DRESSINGS OR ABSORBENT PADS; FIRST-AID KITS
    • A61F13/00—Bandages or dressings; Absorbent pads
    • A61F13/01—Non-adhesive bandages or dressings
    • A61F13/01034—Non-adhesive bandages or dressings characterised by a property
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61F—FILTERS IMPLANTABLE INTO BLOOD VESSELS; PROSTHESES; DEVICES PROVIDING PATENCY TO, OR PREVENTING COLLAPSING OF, TUBULAR STRUCTURES OF THE BODY, e.g. STENTS; ORTHOPAEDIC, NURSING OR CONTRACEPTIVE DEVICES; FOMENTATION; TREATMENT OR PROTECTION OF EYES OR EARS; BANDAGES, DRESSINGS OR ABSORBENT PADS; FIRST-AID KITS
    • A61F13/00—Bandages or dressings; Absorbent pads
    • A61F13/00051—Accessories for dressings
    • A61F13/00063—Accessories for dressings comprising medicaments or additives, e.g. odor control, PH control, debriding, antimicrobic
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61F—FILTERS IMPLANTABLE INTO BLOOD VESSELS; PROSTHESES; DEVICES PROVIDING PATENCY TO, OR PREVENTING COLLAPSING OF, TUBULAR STRUCTURES OF THE BODY, e.g. STENTS; ORTHOPAEDIC, NURSING OR CONTRACEPTIVE DEVICES; FOMENTATION; TREATMENT OR PROTECTION OF EYES OR EARS; BANDAGES, DRESSINGS OR ABSORBENT PADS; FIRST-AID KITS
    • A61F13/00—Bandages or dressings; Absorbent pads
    • A61F13/00987—Apparatus or processes for manufacturing non-adhesive dressings or bandages
    • A61F13/00991—Apparatus or processes for manufacturing non-adhesive dressings or bandages for treating webs, e.g. for moisturising, coating, impregnating or applying powder
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61F—FILTERS IMPLANTABLE INTO BLOOD VESSELS; PROSTHESES; DEVICES PROVIDING PATENCY TO, OR PREVENTING COLLAPSING OF, TUBULAR STRUCTURES OF THE BODY, e.g. STENTS; ORTHOPAEDIC, NURSING OR CONTRACEPTIVE DEVICES; FOMENTATION; TREATMENT OR PROTECTION OF EYES OR EARS; BANDAGES, DRESSINGS OR ABSORBENT PADS; FIRST-AID KITS
    • A61F13/00—Bandages or dressings; Absorbent pads
    • A61F13/01—Non-adhesive bandages or dressings
    • A61F13/01021—Non-adhesive bandages or dressings characterised by the structure of the dressing
    • A61F13/01029—Non-adhesive bandages or dressings characterised by the structure of the dressing made of multiple layers
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/43—Enzymes; Proenzymes; Derivatives thereof
    • A61K38/46—Hydrolases (3)
    • A61K38/48—Hydrolases (3) acting on peptide bonds (3.4)
    • A61K38/482—Serine endopeptidases (3.4.21)
    • A61K38/4833—Thrombin (3.4.21.5)
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L15/00—Chemical aspects of, or use of materials for, bandages, dressings or absorbent pads
    • A61L15/16—Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons
    • A61L15/22—Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons containing macromolecular materials
    • A61L15/225—Mixtures of macromolecular compounds
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L15/00—Chemical aspects of, or use of materials for, bandages, dressings or absorbent pads
    • A61L15/16—Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons
    • A61L15/22—Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons containing macromolecular materials
    • A61L15/32—Proteins, polypeptides; Degradation products or derivatives thereof, e.g. albumin, collagen, fibrin, gelatin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L15/00—Chemical aspects of, or use of materials for, bandages, dressings or absorbent pads
    • A61L15/16—Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons
    • A61L15/42—Use of materials characterised by their function or physical properties
    • A61L15/64—Use of materials characterised by their function or physical properties specially adapted to be resorbable inside the body
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P7/00—Drugs for disorders of the blood or the extracellular fluid
    • A61P7/04—Antihaemorrhagics; Procoagulants; Haemostatic agents; Antifibrinolytic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61F—FILTERS IMPLANTABLE INTO BLOOD VESSELS; PROSTHESES; DEVICES PROVIDING PATENCY TO, OR PREVENTING COLLAPSING OF, TUBULAR STRUCTURES OF THE BODY, e.g. STENTS; ORTHOPAEDIC, NURSING OR CONTRACEPTIVE DEVICES; FOMENTATION; TREATMENT OR PROTECTION OF EYES OR EARS; BANDAGES, DRESSINGS OR ABSORBENT PADS; FIRST-AID KITS
    • A61F13/00—Bandages or dressings; Absorbent pads
    • A61F2013/00089—Wound bandages
    • A61F2013/00106—Wound bandages emergency bandages, e.g. for first aid
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61F—FILTERS IMPLANTABLE INTO BLOOD VESSELS; PROSTHESES; DEVICES PROVIDING PATENCY TO, OR PREVENTING COLLAPSING OF, TUBULAR STRUCTURES OF THE BODY, e.g. STENTS; ORTHOPAEDIC, NURSING OR CONTRACEPTIVE DEVICES; FOMENTATION; TREATMENT OR PROTECTION OF EYES OR EARS; BANDAGES, DRESSINGS OR ABSORBENT PADS; FIRST-AID KITS
    • A61F13/00—Bandages or dressings; Absorbent pads
    • A61F2013/00361—Plasters
    • A61F2013/00365—Plasters use
    • A61F2013/00463—Plasters use haemostatic
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61F—FILTERS IMPLANTABLE INTO BLOOD VESSELS; PROSTHESES; DEVICES PROVIDING PATENCY TO, OR PREVENTING COLLAPSING OF, TUBULAR STRUCTURES OF THE BODY, e.g. STENTS; ORTHOPAEDIC, NURSING OR CONTRACEPTIVE DEVICES; FOMENTATION; TREATMENT OR PROTECTION OF EYES OR EARS; BANDAGES, DRESSINGS OR ABSORBENT PADS; FIRST-AID KITS
    • A61F13/00—Bandages or dressings; Absorbent pads
    • A61F2013/00361—Plasters
    • A61F2013/00365—Plasters use
    • A61F2013/00463—Plasters use haemostatic
    • A61F2013/00472—Plasters use haemostatic with chemical means
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61F—FILTERS IMPLANTABLE INTO BLOOD VESSELS; PROSTHESES; DEVICES PROVIDING PATENCY TO, OR PREVENTING COLLAPSING OF, TUBULAR STRUCTURES OF THE BODY, e.g. STENTS; ORTHOPAEDIC, NURSING OR CONTRACEPTIVE DEVICES; FOMENTATION; TREATMENT OR PROTECTION OF EYES OR EARS; BANDAGES, DRESSINGS OR ABSORBENT PADS; FIRST-AID KITS
    • A61F13/00—Bandages or dressings; Absorbent pads
    • A61F2013/00361—Plasters
    • A61F2013/00365—Plasters use
    • A61F2013/0054—Plasters use for deep wounds
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61F—FILTERS IMPLANTABLE INTO BLOOD VESSELS; PROSTHESES; DEVICES PROVIDING PATENCY TO, OR PREVENTING COLLAPSING OF, TUBULAR STRUCTURES OF THE BODY, e.g. STENTS; ORTHOPAEDIC, NURSING OR CONTRACEPTIVE DEVICES; FOMENTATION; TREATMENT OR PROTECTION OF EYES OR EARS; BANDAGES, DRESSINGS OR ABSORBENT PADS; FIRST-AID KITS
    • A61F13/00—Bandages or dressings; Absorbent pads
    • A61F2013/00361—Plasters
    • A61F2013/00902—Plasters containing means
    • A61F2013/0091—Plasters containing means with disinfecting or anaesthetics means, e.g. anti-mycrobic
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61F—FILTERS IMPLANTABLE INTO BLOOD VESSELS; PROSTHESES; DEVICES PROVIDING PATENCY TO, OR PREVENTING COLLAPSING OF, TUBULAR STRUCTURES OF THE BODY, e.g. STENTS; ORTHOPAEDIC, NURSING OR CONTRACEPTIVE DEVICES; FOMENTATION; TREATMENT OR PROTECTION OF EYES OR EARS; BANDAGES, DRESSINGS OR ABSORBENT PADS; FIRST-AID KITS
    • A61F13/00—Bandages or dressings; Absorbent pads
    • A61F2013/00361—Plasters
    • A61F2013/00902—Plasters containing means
    • A61F2013/00927—Plasters containing means with biological activity, e.g. enzymes for debriding wounds or others, collagen or growth factors
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L2400/00—Materials characterised by their function or physical properties
    • A61L2400/04—Materials for stopping bleeding

Definitions

  • the present invention relates to a hemostatic dressing that comprises a plurality of layers that contain resorbable materials and/or coagulation proteins.
  • the hemostatic dressing is useful for the treatment of wounded tissue.
  • Liquid fibrin sealants have been used as an operating room adjunct to hemorrhage control.
  • the widespread use of fibrinogen and thrombin was common in the last year of World War II, but was abandoned because of the transmission of hepatitis.
  • D. B. Kendrick, Blood Program in WW II (Washington, DC: Office of the Surgeon General, Department of Army; 1989), 363-368.
  • a dry fibrinogen-thrombin dressing (T ACHO C OMB TM, Hafslund Nycomed Pharma, Linz, Austria) is also available for operating room use in many European countries. Schiele et al., Clin. Materials 9:169-177 (1992). Present formulations of this dressing use bovine thrombin. While this fibrinogen-thrombin dressing requires no premixing and is easy to use, its utility is limited by a requirement for storage at 4° C. and the necessity for prewetting with saline solution prior to application to the wound.
  • a hemostatic sandwich dressing which contains a layer of thrombin sandwiched between layers of fibrinogen (see, e.g., PCT/US99/10952, which is incorporated herein by reference).
  • such dressings can be used in methods for treating wounded tissue, such conventional sandwich dressings can become delaminated, whereby the edges of the layers of the dressing no longer adhere to each other. Such delamination can result in reduced interaction of the dressing components layers, with decreased effectiveness of the dressing in preventing hemorrhage.
  • the present invention provides a hemostatic dressing (e.g., a bandage) that includes a layer of thrombin sandwiched between a first and a second layer of fibrinogen, wherein the thrombin layer is noncoextensive with the first and/or second fibrinogen layer.
  • a hemostatic dressing e.g., a bandage
  • Such a hemostatic dressing is useful for treating wounds and offers the unexpected advantage of inhibiting delamination of the layers, as compared with dressings in which the thrombin layer is coextensive with the entire first and second fibrinogen layers.
  • the present invention provides a hemostatic dressing which comprises: (i) a first fibrinogen layer; (ii) a thrombin layer adjacent to the first fibrinogen layer; and (iii) a second fibrinogen layer adjacent to the thrombin layer, wherein the thrombin layer is non-coextensive with the first and/or second fibrinogen layers of the hemostatic dressing.
  • a related dressing of the invention comprises: (i) a resorbable material layer; (ii) a first fibrinogen layer adjacent to the resorbable material layer; (iii) a thrombin layer adjacent to the first fibrinogen layer; and (iv) a second fibrinogen layer adjacent to the thrombin layer, wherein the thrombin layer is noncoextensive with the first and/or second fibrinogen layers of the hemostatic dressing.
  • the hemostatic dressing comprises: (i) a first fibrinogen layer; (ii) a resorbable material layer adjacent to the first fibrinogen layer; (iii) a thrombin layer adjacent to the resorbable material layer; and (iv) a second fibrinogen layer adjacent to the thrombin layer, wherein the thrombin layer is non-coextensive with the first and/or second fibrinogen layers of the hemostatic dressing.
  • the invention also includes methods for treating wounded tissue in a patient, which comprise applying any of the novel hemostatic dressings described herein to wounded tissue.
  • the hemostatic dressing can be hydrated with liquids that are exogenous to the wounded tissue, or they can be hydrated with liquids that are endogenous to the wounded tissue.
  • the invention includes a method for preparing a hemostatic dressing by providing a resorbable or nonresorbable backing layer having attached thereto a first layer of fibrinogen; applying a layer of thrombin to said first layer of fibrinogen on a side of the fibrinogen layer that is opposite of the side to which the resorbable or nonresorbable backing layer is attached; and applying a second layer of fibrinogen to the layer of thrombin, wherein the layer of thrombin is noncoextensive with the first fibrinogen layer and/or noncoextensive with the second fibrinogen layer.
  • the thrombin layer is coextensive with 5% to 95% (e.g., 20 to 50%) of the first and second fibrinogen layers, independently.
  • the thrombin layer can be configured in any of a variety of shapes and patterns.
  • the thrombin layer can be configured as an array of spots comprising thrombin, or as a single spot comprising thrombin.
  • the thrombin layer can be configured as a plurality of lines comprising thrombin.
  • Each layer of the hemostatic dressings can also optionally contain one or more suitable fillers, binding agents and/or solubilizing agents.
  • each of the hemostatic dressings can also optionally further comprise a release layer which contains a release agent and/or a backing material.
  • a thrombin layer that is said to be “noncoextensive” with a fibrinogen layer is one in which the spatial boundaries of the thrombin layer in two dimensions are smaller than the spatial boundaries of one or both fibrinogen layers such that the thrombin layer is coextensive with only about 5% to about 95% of the surface area of the first fibrinogen layer of the hemostatic dressing and/or coextensive with only about 5% to about 95% of the surface layer of the second fibrinogen layer of the hemostatic dressing, independently.
  • the thrombin layer can be coextensive with about 10, 20, 30, 40, 50, 60, 70, 75, 80, or 90% of the surface area of each of the first and second fibrinogen layers, independently.
  • a thrombin layer that is “coextensive” with a fibrinogen layer provides full coverage of the fibrinogen layer and is coextensive with 100% of the surface area of the fibrinogen layer.
  • a thrombin layer can be noncoextensive with the first fibrinogen layer and yet be coextensive with the second fibrinogen layer, or vice versa, e.g., by employing fibrinogen layers having different total surface areas or shapes.
  • Patient refers to human or animal individuals in need of medical care and/or treatment.
  • “Wound” as used herein refers to any damage to any tissue of a patient that results in the loss of blood from the circulatory system.
  • the tissue can be an internal tissue, such as an organ or blood vessel, or an external tissue, such as the skin.
  • the loss of blood can be internal, such as from a ruptured organ, or external, such as from a laceration.
  • a wound can be in a soft tissue, such as an organ, or in hard tissue, such as bone.
  • the damage may have been caused by any agent or source, including traumatic injury, infection or surgical intervention. The damage can be life-threatening or non-life-threatening.
  • Resorbable material refers to a material that is broken down spontaneously and/or by the mammalian body into components which are consumed or eliminated in such a manner as not to interfere significantly with wound healing and/or tissue regeneration, and without causing any significant metabolic disturbance.
  • “Stability” as used herein refers to the retention of those characteristics of a material that determine activity and/or function.
  • Blooding agent refers to a compound or mixture of compounds that improves the adherence of one layer of the hemostatic dressing to one or more different layers and/or the adherence of the components of a given layer to other components of that layer.
  • Solubilizing agent refers to a compound or mixture of compounds that improves the dissolution of a protein or proteins in aqueous solvent.
  • Fill refers to a compound or mixture of compounds that provide bulk and/or porosity to one or more layers of the hemostatic dressings.
  • Release agent refers to a compound or mixture of compounds that facilitates removal of an hemostatic dressing from a manufacturing mold.
  • “Foaming agent” as used herein refers to a compound or mixture of compounds that produces gas when hydrated under suitable conditions.
  • the hemostatic dressing of the invention offers various advantages as compared with conventional dressings.
  • the dressings of the invention are less likely to become delaminated at their edges, thus rendering the dressings more durable and easier to handle than conventional dressings.
  • such dressings are more amenable to large-scale manufacturing and provide for better control of the amount of thrombin dispensed in the dressing.
  • An exemplary embodiment of the present invention is directed to a hemostatic dressing, e.g., for treating wounded tissue in a patient, which comprises: (i) a first fibrinogen layer; (ii) a thrombin layer adjacent to the first fibrinogen layer; and (iii) a second fibrinogen layer adjacent to the thrombin layer, wherein the thrombin layer is noncoextensive with the first and/or second fibrinogen layers.
  • Another embodiment of the present invention is directed to a hemostatic dressing which comprises: (i) a resorbable material layer; (ii) a first fibrinogen layer adjacent to the resorbable material layer; (iii) a thrombin layer adjacent to the first fibrinogen layer; and (iv) a second fibrinogen layer adjacent to the thrombin layer, wherein the thrombin layer is noncoextensive with the first and/or second fibrinogen layers.
  • Yet another embodiment of the present invention is directed to a hemostatic dressing for treating wounded tissue in a patient which comprises: (i) a first fibrinogen layer; (ii) a resorbable material layer adjacent to the first fibrinogen layer; (iii) a thrombin layer adjacent to the resorbable material layer; and (iv) a second fibrinogen layer adjacent to the thrombin layer, wherein the thrombin layer is noncoextensive with the first and/or second fibrinogen layers.
  • Each layer of the hemostatic dressings can also optionally contain one or more suitable fillers, such as sucrose.
  • Each layer of the hemostatic dressings can also optionally contain one or more suitable binding agents, such as sucrose.
  • Each layer of the hemostatic dressings can also optionally contain one or more suitable solubilizing agents, such as sucrose.
  • Each layer of the hemostatic dressings can also optionally contain one or more suitable foaming agents, such as a mixture of citric acid and sodium bicarbonate.
  • Each of the hemostatic dressings can also optionally further comprise a release layer which contains a release agent.
  • a release agent is sucrose.
  • Each of the hemostatic dressings can also further comprise a backing material on the side of the dressing opposite the wound-facing side when the dressing is in use.
  • the backing material can be affixed with a physiologically-acceptable adhesive or can be self-adhering (e.g. by having a surface static charge).
  • the backing material can be a resorbable material or a non-resorbable material, such as a silicone patch or plastic.
  • the fibrinogen employed in the hemostatic dressing can be a fibrinogen complex or any fibrinogen, or a derivative or metabolite thereof (such as fibrinopeptide A and fibrinopeptide B) can be employed as desired.
  • the fibrinogen can also contain Factor XIII.
  • the fibrinogen complex can be a mixture of human plasma proteins which has been purified and virally inactivated.
  • An exemplary aqueous solution of fibrinogen complex contains 100-130 mg/mL total protein, of which at least 80% is fibrinogen.
  • Other constituents of the fibrinogen complex can include albumin (generally about 5-25 mg/mL); plasminogen (generally less than about 5 ⁇ g/mL); Factor XIII (generally about 10-40 Units/mL); and polysorbate 80 (generally less than 3%).
  • the pH of the fibrinogen complex is generally in the range of 7.1-7.5. Suitable fibrinogen complexes can also contain fibronectin.
  • the fibrinogen applied to form a layer of the dressing typically has a concentration of 1 mg/cm 2 to 60 mg/cm 2 , e.g., at least 5, 10, 15, 20, 30, 40, 50 mg/cm 2 .
  • the first and second fibrinogen layers can be the same size, e.g., such that the second fibrinogen layer generally is coextensive with the first fibrinogen layer.
  • the first fibrinogen layer can be noncoextensive with the second fibrinogen layer; or the second fibrinogen layer can be noncoextensive with the first fibrinogen layer.
  • the first fibrinogen layer can be up to 100% of the size of the second fibrinogen layer, or the second fibrinogen layer can be up to 100% of the size of the first fibrinogen layer.
  • the thrombin employed in the hemostatic dressing can be a lyophilized mixture of human plasma proteins which have been purified and virally inactivated.
  • the dressings of the invention typically contain thrombin at a potency of about 1 to 160 International Units (IU)/cm 2 , e.g., at least 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 60, 70, 80, 100, 125, 150 IU/cm 2 .
  • Optional constituents include albumin (generally about 5 mg/mL) and glycine (generally about 0.3 M ⁇ 0.05M).
  • the pH of the thrombin is generally in the range of 6.5-7.1.
  • the thrombin layer is applied to the first fibrinogen layer such that it is noncoextensive with the first fibrinogen layer and/or will be noncoextensive with the second fibrinogen layer upon application of the second fibrinogen layer.
  • the thrombin layer can occupy about 5% to about 95% of the surface area of the first fibrinogen layer and/or about 5% to about 95% of the surface area of the second fibrinogen layer.
  • the thrombin can be applied to the fibrinogen layer in a single spot or as a series of spots on the fibrinogen layer such that the total surface area of the thrombin spots occupies about 5% to about 95% of the surface area of the first fibrinogen layer and/or about 5% to about 95% of the surface area of the second fibrinogen layer.
  • Such a spot or spots of thrombin can have any geometric shape, e.g., filled or unfilled circles, rectangles, triangles, lines, amorphous shapes, or combinations thereof. Such spots can be applied to the first fibrinogen layer in an ordered or random pattern.
  • a plurality of spots can form any of a variety of shapes and patterns, such as an array, a grid, a series of concentric spots (e.g., concentric circles or squares), an overlapping series of spots (e.g., overlapping circles), spokes emanating from an axis, or any other configuration, provided that the total surface area of the thrombin is about 5% to about 95% of the surface area of the first fibrinogen layer and/or about 5% to about 95% of the surface area of the second fibrinogen layer.
  • a large number of small spots is preferred over a small number of large spots.
  • a 20 ⁇ 20 array of spots generally is preferable over a 10 ⁇ 10 array of spots occupying the same total surface area.
  • the spots can be of any size provided that the total surface area of the thrombin is about 5% to about 95% of the surface area of the first fibrinogen layer and/or about 5% to about 95% of the surface area of the second fibrinogen layer.
  • the spots can be, without limitation, at least about 0.01, 0. 1, 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 mm or more in diameter, width, or length.
  • 4 circular spots having a diameter of 2-3 mm each can occupy a square centimeter of a dressing.
  • a variety of other configurations are within the scope of the invention and can readily be utilized by those skilled in the art.
  • the dressing can be prepared as any of a variety of sizes and shapes.
  • the dressings are of a size and shape that can readily be handled by those skilled in the art, typically less than 12′′ in length along any side, e.g., 1′′ ⁇ 1′′, 1′′ ⁇ 2′′, 4′′ ⁇ 4′′, etc.
  • the moisture level of the dressing typically is less than 8% (e.g., less than 7, 6, 5, 4, 3, 2, or 1%).
  • the resorbable material can be a proteinaceous substance, such as fibrin, keratin, collagen and/or gelatin, or a carbohydrate substances, such as alginates, chitin, cellulose, proteoglycans (e.g. poly-N-acetyl glucosamine), glycolic acid polymers, lactic acid polymers, or glycolic acid/lactic acid co-polymers.
  • the resorbable material can be a carbohydrate substance.
  • Illustrative examples of resorbable materials are sold under the tradenames VICRYLTM and DEXONTM.
  • the various layers of the hemostatic dressing can be affixed to one another by any means known and available to those skilled in the art.
  • the fibrinogen layer(s) and/or the thrombin layer(s) is (are) applied as a series of quick-frozen aqueous solution layers and subsequently lyophilized or freeze-dried, e.g., after application of each layer, and upon assembly of the entire dressing.
  • the layers can be applied by any of a variety of techniques, including spraying, pipetting (e.g., with a multi-channel pipettor), sprinkling, using a mask, electrostatic deposition, using a microsyringe array system, or dispensing using a dispensing manifold that contains ports for producing a high density array.
  • the hemostatic dressing when the dressings are prepared using a mold, a release agent, such as sucrose, is applied to the mold before the first layer of the dressing is applied.
  • a release agent such as sucrose
  • the hemostatic dressing further comprises a release layer, which contains said release agent.
  • a physiologically-acceptable adhesive can be applied to the resorbable material and/or the backing material (when present) and the fibrinogen layer(s) and/or the thrombin layer(s) subsequently affixed thereto.
  • the physiologically-acceptable adhesive has a shear strength and/or structure such that the resorbable material and/or backing material can be separated from the fibrinogen layer after application of the dressing to wounded tissue.
  • the physiologically-acceptable adhesive has a shear strength such that the resorbable material and/or backing material cannot be separated from the fibrinogen layer after application of the dressing to wounded tissue.
  • Suitable fibrinogen and thrombin can be obtained from human or mammalian plasma by any of the purification methods known and available to those skilled in the art; from supernatants or pastes of recombinant tissue culture, viruses, yeast, bacteria, or the like that contain a gene that expresses a human or mammalian plasma protein which has been introduced according to standard recombinant DNA techniques; or from the fluids (e.g., blood, milk, lymph, urine or the like) of transgenic animals that contain a gene that expresses human fibrinogen and/or human thrombin which has been introduced according to standard transgenic techniques.
  • the fluids e.g., blood, milk, lymph, urine or the like
  • the purity of the fibrinogen and/or the thrombin for use in the hemostatic dressing will be of an appropriate purity known to one of ordinary skill in the relevant art to lead to efficacy and stability of the protein.
  • the fibrinogen and/or the thrombin can be subjected to multiple chromatographic purification steps, such as affinity chromatography and immunoaffinity chromatography, to remove substances which may cause fragmentation, activation and/or degradation of the fibrinogen and/or the thrombin during manufacture, storage and/or use.
  • Illustrative examples of such substances that can be removed by purification include protein contaminants, such as plasminogen, inter-alpha trypsin inhibitor and pre-alpha trypsin inhibitor; non-protein contaminants, such as lipids; and mixtures of protein and non-protein contaminants, such as lipoproteins.
  • protein contaminants such as plasminogen, inter-alpha trypsin inhibitor and pre-alpha trypsin inhibitor
  • non-protein contaminants such as lipids
  • mixtures of protein and non-protein contaminants such as lipoproteins.
  • the fibrinogen and the thrombin can be activated at the time the dressing is applied to the wounded tissue by the endogenous fluids (e.g., blood) of the patient escaping from the hemorrhaging wound.
  • the fibrinogen and or the thrombin can be activated by a application of a physiologically-acceptable liquid (e.g., water, buffer, saline), optionally containing any necessary co-factors and/or enzymes, prior to or upon application of the hemostatic dressing to the wounded tissue.
  • a physiologically-acceptable liquid e.g., water, buffer, saline
  • one or more supplements can also be contained in one or more layers of the hemostatic dressing, e.g., drugs such as growth factors, polyclonal and monoclonal antibodies and other compounds.
  • antibiotics such as tetracycline and ciprofloxacin, amoxicillin, and metronidazole
  • anticoagulants such as activated protein C, heparin, prostracyclin (PGI 2 ), prostaglandins, leukotrienes, antithrombin III, ADPase, and plasminogen activator
  • steroids such as dexamethasone, inhibitors of prostacyclin, prostaglandins, leukotrienes and/or kinins to inhibit inflammation
  • cardiovascular drugs such as calcium channel blockers, vasodilators and vasoconstrictors; chemoattractants; local anesthetics such as bupivacaine; and antiproliferative/antitumor
  • illustrative supplements include, but are not limited to: vitamins and other nutritional supplements; glycoproteins; fibronectin; peptides and proteins; carbohydrates (both simple and/or complex); proteoglycans; antiangiogenins; antigens; lipids or liposomes; and oligonucleotides (sense and/or antisense DNA and/or RNA).
  • the example set forth below demonstrates that delamination of the dressings can be decreased by preparing a dressing in which the thrombin layer is not coextensive with the first and second fibrinogen layers.
  • thrombin was dispensed onto the first fibrinogen layer in either of two configurations. In the conventional dressings, the thrombin fully covered the first fibrinogen layer (this configuration is referred to herein as “full” coverage). In the dressings of the invention, the thrombin was configured on top of the first fibrinogen layer as a single circle that was not coextensive with the first fibrinogen layer (this configuration is referred to herein as “circle” coverage).
  • fibrinogen was formulated in a conventional manner: 35 mg/ml of total protein (TP) in Buffer D (100 mM NaCl, 1.1 mM CaCl 2 .H 2 O, 10 mM TrisHCl, 10 mM Sodium Citrate, 2% Sucrose, 2.8 mg/ml albumin, 0.52 mg/ml TWEEN-80, pH 7.2) containing albumin at 80 mg/g of TP and polysorbate at 15 mg/g of TP.
  • Thrombin concentrate having a potency of 4745 IU/ml, was formulated in each of four different buffers to obtain a thrombin solution containing 2000 IU/ml. The four buffers are described in Table 1.
  • the dressings were manufactured manually, and 18 dressings were produced for each of the four thrombin formulations and coverage types to produce a total of 144 dressings.
  • the dressings were freeze-dried, packaged with desiccant, then tested in in vitro assays for appearance, moisture content, gamma dimerization, and percent clottable protein, and in an ex vivo pig arteriotomy assay.
  • dressings were tested for delamination in an appearance assay. Dressings were considered to be passing if the fibrinogen layers of the dressing were attached to each other along all four edges. Delamination of the dressing layers makes the dressings difficult to handle and renders the dressings susceptible to fragmentation if surface cracks in the layers are also present. As shown in Table 1, dressings in which the thrombin layer was applied with circle coverage (groups 2, 4, 6, and 8) had a higher pass rate in the delamination test (i.e., appearance test) than did dressings in which the thrombin layer was applied with full coverage (groups 1, 3, 5, and 7).
  • the moisture content was measured for one of the dressings from each group.
  • the moisture content of the dressings averaged 0.98% and was very consistent throughout the groups, ranging from 0.80 to 1.16%. This moisture content level is typical for such dressings.
  • a gamma-gamma dimer assay measures the ability of thrombin to cross-link the gamma chains of fibrinogen, thus forming a clot.
  • each dressing is ground into a uniform, fine powder in a low moisture chamber and divided into two equal portions.
  • ODS Okude Dissolving Solution
  • a standard ex vivo porcine arteriotomy assay can be performed as follows. Obtain frozen porcine aorta and thaw. Aortas can be thawed overnight at 4° C., or individually wrapped in the water bath at 37° C. Dissect excess connective tissue from approximately first 11 cm of the aorta. Usually, the first 5-5.5 cm are free from collateral vessels. The next 5-5.5 cm should not have more than 1-2 collaterals. These can be easily sealed or patched with cyanoacrylate glue.
  • both O-rings carefully secure over the top of the vessel.
  • the distance between both O-rings should be 3.5 cm.
  • the artery should be snug fitting and held securely in place.
  • Position the secured vessel such that the hole in the syringe lies in the middle of the distance between the O-rings.
  • the flow rate is escalated to approximately 3 mL/min (10 ⁇ speed, both settings). This should be done until a pressure of 200 mm Hg is obtained. Once 200 mm Hg is achieved, start the timer for 2 minutes.
  • a dressing is considered passing if it maintains a fairly consistent pressure of 200 mm Hg for two minutes with absolutely no leakage.
  • a dressing is also considered passing if it maintains a fairly consistent pressure of 200 mm Hg for two minutes with only minimal leakage (e.g., slow seeping or a leak that has resealed itself).
  • a dressing is considered failing if it cannot maintain adequate pressure due to severe leakage. This includes leakage caused by poor adhesion, as well as leakage due to manufacturing flaws. Dressings that may be considered as failing in this assay can nonetheless be used to treat less severe wounds.
  • fibrinogen was formulated as described above, and thrombin was formulated as described in Table 2.
  • the fibrinogen layers (approx 1.2 mL) were applied using a programmable pipette, and the dressings were manufactured manually as described herein.
  • the middle layer of the dressing i.e., thrombin or fibrinogen
  • the middle layer of the dressing was applied either by spraying using an air brush or by pipetting.
  • four different volumes of the middle layer were applied, with 120 ⁇ l considered to be a “standard” volume.
  • the spraying time was increased to increase the volume applied. Full coverage of the first fibrinogen layer was achieved with all groups in which the thrombin was sprayed.

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