US20080033332A1 - Solid dressing for treating wounded tissue - Google Patents

Solid dressing for treating wounded tissue Download PDF

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Publication number
US20080033332A1
US20080033332A1 US11/882,876 US88287607A US2008033332A1 US 20080033332 A1 US20080033332 A1 US 20080033332A1 US 88287607 A US88287607 A US 88287607A US 2008033332 A1 US2008033332 A1 US 2008033332A1
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US
United States
Prior art keywords
fibrinogen
thrombin
dressing
solid dressing
solid
Prior art date
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
Application number
US11/882,876
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English (en)
Inventor
Martin MacPhee
Dawson Beall
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
STB LIFESAVING TECHNOLOGIES Inc
Original Assignee
STB LIFESAVING TECHNOLOGIES Inc
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by STB LIFESAVING TECHNOLOGIES Inc filed Critical STB LIFESAVING TECHNOLOGIES Inc
Priority to US11/882,876 priority Critical patent/US20080033332A1/en
Assigned to STB LIFESAVING TECHNOLOGIES, INC. reassignment STB LIFESAVING TECHNOLOGIES, INC. ASSIGNMENT OF ASSIGNORS INTEREST (SEE DOCUMENT FOR DETAILS). Assignors: BEALL, DAWSON, MACPHEE, MARTIN
Publication of US20080033332A1 publication Critical patent/US20080033332A1/en
Priority to US13/363,489 priority patent/US20120148658A1/en
Assigned to STB, LTD reassignment STB, LTD CORRECTIVE ASSIGNMENT TO CORRECT THE NAME OF ASSIGNEE PREVIOUSLY RECORDED ON REEL 020027 FRAME 0230. ASSIGNOR(S) HEREBY CONFIRMS THE ASSIGNMENT OF ASSIGNOR'S INTEREST. Assignors: BEALL, DAWSON, MACPHEE, MARTIN
Priority to US14/599,519 priority patent/US20150132363A1/en
Priority to US15/208,591 priority patent/US20160317355A1/en
Priority to US15/605,660 priority patent/US20180117210A1/en
Priority to US16/289,535 priority patent/US20200000957A1/en
Abandoned legal-status Critical Current

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    • A61L2300/40—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a specific therapeutic activity or mode of action
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    • A61L2400/04—Materials for stopping bleeding

Definitions

  • the present invention relates to a solid dressing for treating wounded tissue in a mammalian patient, such as a human.
  • haemostatic agents that have been developed to overcome the deficiencies of traditional gauze bandages. These haemostatic agents include the following:
  • TraumaDEXTM a starch-based haemostatic agent sold under the trade name TraumaDEXTM.
  • TraumaDEXTM This product comprises microporous polysaccharide particles that are poured directly into or onto a wound. The particles appear to exert their haemostatic effect by absorbing water from the blood and plasma in the wound, resulting in the accumulation and concentration of clotting factors and platelets.
  • this agent showed no meaningful benefit over standard gauze dressings. See McManus et al., Business Briefing: Emergency Medical Review 2005, pp. 76-79 (presently available on-line at www.touchbriefings.com/pdf/1334/Wedmore.pdf).
  • QuickClotTM powder a zeolite granular haemostatic agent that is poured directly into or onto a wound.
  • the zeolite particles also appear to exert their haemostatic effect through fluid absorption, which cause the accumulation and concentration of clotting factors and platelets.
  • this agent has been used successfully in some animal studies, there remains concern about the exothermic process of fluid absorption by the particles. Some studies have shown this reaction to produce temperatures in excess of 143° C. in vitro and in excess of 50° C. in vivo, which is severe enough to cause third-degree burns. See McManus et al., Business Briefing: Emergency Medical Review 2005, at 77.
  • the exothermic reaction of QuikClotTM has also been observed to result in gross and histological tissue changes of unknown clinical significance. Acheson et al., J. Trauma 59:865-874 (2005).
  • the Rapid Deployment HemostatTM appears to exert its haemostatic effect through red blood cell aggregation, platelet activation, clotting cascade activation and local vasoconstriction.
  • the Rapid Deployment HemostatTM is an algae-derived dressing composed of poly-N-acetyl-glucosamine. While the original dressing design was effective in reducing minor bleeding, it was necessary to add gauze backing in order to reduce blood loss in swine models of aortic and liver injury. See McManus et al., Business Briefing: Emergency Medical Review 2005, at 78.
  • HemConTM Chitosan Bandage is a freeze-dried chitosan dressing purportedly designed to optimize the mucoadhesive surface density and structural integrity of the chitosan at the site of the wound.
  • the HemConTM Chitosan Bandage apparently exerts its haemostatic effects primarily through adhesion to the wound, although there is evidence suggesting it may also enhance platelet function and incorporate red blood cells into the clot it forms on the wound.
  • This bandage has shown improved hemostasis and reduced blood loss in several animal models of arterial hemorrhage, but a marked variability was observed between bandages, including the failure of some due to inadequate adherence to the wound. See McManus et al., Business Briefing: Emergency Medical Review 2005, at 79.
  • Liquid fibrin sealants such as Tisseel VH have been used for years as an operating room adjunct for hemorrhage control. See J. L. Garza et al., J. Trauma 30:512-513 (1990); H. B. Kram et al., J. Trauma 30:97-101 (1990); M. G. Ochsner et al., J. Trauma 30:884-887 (1990); T. L. Matthew et al., Ann. Thorac. Surg. 50:40-44 (1990); H. Jakob et al., J. Vasc. Surg., 1:171-180 (1984). The first mention of tissue glue used for hemostasis dates back to 1909.
  • Liquid fibrin sealants are typically composed of fibrinogen and thrombin, but may also contain Factor XIII/XIIIa, either as a by-product of fibrinogen purification or as an added ingredient (in certain applications, it is therefore not necessary that Factor XIII/Factor XIIIa be present in the fibrin sealant because there is sufficient Factor XIII/XIIIa, or other transaminase, endogenously present to induce fibrin formation). As liquids, however, these fibrin sealants have not proved useful for treating traumatic injuries in the field.
  • Dry fibrinogen-thrombin dressings having a collagen support are also available for operating room use in many European countries. See U. Schiele et al., Clin. Materials 9:169-177 (1992). While these fibrinogen-thrombin dressings do not require the pre-mixing needed by liquid fibrin sealants, their utility for field applications is limited by a requirement for storage at 4° C. and the necessity for pre-wetting with saline solution prior to application to the wound. These dressings are also not effective against high pressure, high volume bleeding. See Sondeen et al., J. Trauma 54:280-285 (2003).
  • a dry fibrinogen/thrombin dressing for treating wounded tissue is also available from the American Red Cross (ARC).
  • ARC American Red Cross
  • this particular dressing is composed of a backing material and a plurality of layers, the outer two of which contain fibrinogen (but no thrombin) while the inner layer contains thrombin and calcium chloride (but no fibrinogen). While this dressing has shown great success in several animal models of hemorrhage, the bandage is fragile, inflexible, and has a tendency to break apart when handled. See McManus et al., Business Briefing: Emergency Medical Review 2005, at 78; Kheirabadi et al., J. Trauma 59:25-35 (2005). In addition, U.S. Pat. No.
  • U.S. Pat. No. 4,683,142 discloses a resorptive sheet material for closing and healing wounds which consists of a glycoprotein matrix, such as collagen, containing coagulation proteins, such as fibrinogen and thrombin.
  • U.S. Pat. No. 5,702,715 discloses a reinforced biological sealant composed of separate layers of fibrinogen and thrombin, at least one of which also contains a reinforcement filler such as PEG, PVP, BSA, mannitol, FICOLL, dextran, myo-inositol or sodium chlorate.
  • a reinforcement filler such as PEG, PVP, BSA, mannitol, FICOLL, dextran, myo-inositol or sodium chlorate.
  • 6,056,970 discloses dressings composed of a bioabsorbable polymer, such as hyaluronic acid or carboxymethylcellulose, and a haemostatic composition composed of powdered thrombin and/or powdered fibrinogen.
  • U.S. Pat. No. 7,189,410 discloses a bandage composed of a backing material having thereon: (i) particles of fibrinogen; (ii) particles of thrombin; and (iii) calcium chloride.
  • U.S. Patent Application Publication No. US 2006/0155234 A1 discloses a dressing composed of a backing material and a plurality of fibrinogen layers which have discrete areas of thrombin between them. To date, none of these dressings have been approved for use or are available commercially.
  • a first embodiment of the present invention is direct to a solid dressing for treating wounded tissue in a mammal comprising at least one haemostatic layer consisting essentially of fibrinogen and a fibrinogen activator, wherein the fibrinogen is present in an amount between about 3.0 mg/cm 2 of the surface area of the wound facing side of the dressing and 13.0 mg/cm 2 of the surface area of the wound facing side of the dressing.
  • Another embodiment is directed to a method of treating wounded tissue using a solid dressing comprising at least one haemostatic layer consisting essentially of fibrinogen and a fibrinogen activator, wherein the fibrinogen is present in an amount between about 11.0 mg/cm 2 of the surface area of the wound facing side of the dressing and 13.0 mg/cm 2 of the surface area of the wound facing side of the dressing.
  • FIGS. 1A-1C are graphs showing the results achieved in Example 1.
  • FIG. 2 is a diagram of the set-up for the ex vivo porcine arteriotomy assay described herein.
  • 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 which results in the loss of blood from the circulatory system and/or any other fluid from the patient's body.
  • the tissue may be an internal tissue, such as an organ or blood vessel, or an external tissue, such as the skin.
  • the loss of blood may be internal, such as from a ruptured organ, or external, such as from a laceration.
  • a wound may 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.
  • 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.
  • Suitable as used herein is intended to mean that a material does not adversely affect the stability of the dressings or any component thereof.
  • Binding agent refers to a compound or mixture of compounds that improves the adherence and/or cohesion of the components of the haemostatic layer(s) of the dressings.
  • 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 the haemostatic layer(s) of a dressing.
  • Release agent refers to a compound or mixture of compounds that facilitates removal of a 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.
  • Solid as used herein is intended to mean that the dressing will not substantially change in shape or form when placed on a rigid surface, wound-facing side down, and then left to stand at room temperature for 24 hours.
  • a first preferred embodiment of the present invention is directed to a solid dressing for treating wounded tissue in a patient which comprises a haemostatic layer consisting of fibrinogen and a fibrinogen activator, wherein the fibrinogen is present in an amount between 3.0 mg/cm 2 of the surface area of the wound facing side of the dressing and 13.0 mg/cm 2 of the surface area of the wound facing side of the dressing, all values being ⁇ 0.09 mg/cm 2 .
  • the fibrinogen and the fibrinogen activator are the only necessary and essential ingredients of the haemostatic layer(s) of the solid dressing when it is used as intended to treat wounded tissue.
  • the haemostatic layer may contain other ingredients in addition to the fibrinogen and the fibrinogen activator as desired for a particular application, but these other ingredients are not required for the solid dressing to function as intended under normal conditions, i.e. these other ingredients are not necessary for the fibrinogen and fibrinogen activator to react and form enough fibrin to reduce the flow of blood and/or fluid from normal wounded tissue when that dressing is applied to that tissue under the intended conditions of use.
  • the appropriate additional components such as Factor XIII/XIIIa or some other transaminase, may be added to the haemostatic layer(s) without deviating from the spirit of the present invention.
  • the solid dressing of the present invention may contain one or more of these haemostatic layers as well as one or more other layers, such as one or more support layers (e.g. a backing material or an internal support material) and release layers.
  • inventions include similar solid dressings wherein the fibrinogen is present in an amount between 11.0 mg/cm 2 of the surface area of the wound facing side of the dressing and 13.0 mg/cm 2 of the surface area of the wound facing side of the dressing, all values being ⁇ 0.09 mg/cm 2 .
  • Other preferred embodiments include similar solid dressings wherein the fibrinogen is present in an amount between 3.0 mg/cm 2 and 9.0 mg/cm 2 Still other preferred embodiments are directed to similar solid dressings wherein the amount of fibrinogen is: 3.0 mg/cm 2 of the surface area of the wound facing side of the dressing; 5.0 mg/cm 2 ; 7.0 mg/cm 2 ; 9.0 mg/cm 2 ; 11.0 mg/cm 2 ; or 13.0 mg/cm 2 (all values being ⁇ 0.09 mg/cm 2 ).
  • Another preferred embodiment of the present invention is directed to a method for treating wounded tissue in a mammal, comprising placing a solid dressing of the present invention to wounded tissue and applying sufficient pressure to the dressing for a sufficient time for enough fibrin to form to reduce the loss of blood and/or other fluid from the wound.
  • the haemostatic layer(s) of the solid dressing is formed or cast as a single piece.
  • the haemostatic layer is made or formed into or from a single source, e.g. an aqueous solution containing a mixture of the fibrinogen and the fibrinogen activator.
  • the haemostatic layer(s) is preferably substantially homogeneous throughout.
  • the haemostatic layer(s) of the solid dressing are composed of a plurality of particles, each of which consists essentially of fibrinogen and a fibrinogen activator.
  • the haemostatic layer may also contain a binding agent to facilitate or improve the adherence of the particles to one another and/or to any support layer(s).
  • binding agents include, but are not limited to, sucrose, mannitol, sorbitol, gelatin, hyaluron and its derivatives, such as hyaluronic acid, maltose, povidone, starch, chitosan and its derivatives, and cellulose derivatives, such as carboxymethylcellulose, as well as mixtures of two or more thereof.
  • the haemostatic layer(s) of the solid dressing may also optionally contain one or more suitable fillers, such as sucrose, lactose, maltose, silk, fibrin, collagen, albumin, hyaluron and its derivatives, such as hyaluronic acid, polysorbate (TweenTM), chitin, chitosan and its derivatives, such as NOCC-chitosan, alginic acid and salts thereof, cellulose and derivatives thereof, proteoglycans, glycolic acid polymers, lactic acid polymers, glycolic acid/lactic acid co-polymers, and mixtures of two or more thereof.
  • suitable fillers such as sucrose, lactose, maltose, silk, fibrin, collagen, albumin, hyaluron and its derivatives, such as hyaluronic acid, polysorbate (TweenTM), chitin, chitosan and its derivatives, such as NOCC-chitosan, alginic acid and salt
  • the haemostatic layer of the solid dressing may also optionally contain one or more suitable solubilizing agents, such as sucrose, dextrose, mannose, trehalose, mannitol, sorbitol, albumin, hyaluron and its derivatives, such as hyaluronic acid, sorbate, polysorbate (TweenTM), sorbitan (SPANTM) and mixtures of two or more thereof.
  • suitable solubilizing agents such as sucrose, dextrose, mannose, trehalose, mannitol, sorbitol, albumin, hyaluron and its derivatives, such as hyaluronic acid, sorbate, polysorbate (TweenTM), sorbitan (SPANTM) and mixtures of two or more thereof.
  • the haemostatic layer of the solid dressing may also optionally contain one or more suitable foaming agents, such as a mixture of a physiologically acceptable acid (e.g. citric acid or acetic acid) and a physiologically suitable base (e.g. sodium bicarbonate or calcium carbonate).
  • suitable foaming agents include, but are not limited to, dry particles containing pressurized gas, such as sugar particles containing carbon dioxide (see, e.g., U.S. Pat. No. 3,012,893) or other physiologically acceptable gases (e.g. Nitrogen or Argon), and pharmacologically acceptable peroxides.
  • the haemostatic layer(s) of the solid dressing may also optionally contain a suitable source of calcium ions, such as calcium chloride, and/or a fibrin cross-linker, such as a transaminase (e.g. Factor XIII/XIIIa) or glutaraldehyde.
  • a suitable source of calcium ions such as calcium chloride
  • a fibrin cross-linker such as a transaminase (e.g. Factor XIII/XIIIa) or glutaraldehyde.
  • the haemostatic layer of the solid dressing is preferably prepared by mixing aqueous solutions of the fibrinogen and the fibrinogen activator under conditions which minimize the activation of the fibrinogen by the fibrinogen activator.
  • the mixture of aqueous solutions is then subjected to a process such as lyophilization or free-drying to reduce the moisture content to the desired level, i.e. to a level where the dressing is solid and therefore will not substantially change in shape or form upon standing, wound-facing surface down, at room temperature for 24 hours.
  • a process such as lyophilization or free-drying to reduce the moisture content to the desired level, i.e. to a level where the dressing is solid and therefore will not substantially change in shape or form upon standing, wound-facing surface down, at room temperature for 24 hours.
  • Similar processes that achieve the same result such as drying, spray-drying, vacuum drying and vitrification, may also be employed.
  • moisture content refers to the amount freely-available water in the dressing. “Freely-available” means the water is not bound to or complexed with one or more of the non-liquid components of the dressing.
  • the moisture content referenced herein refers to levels determined by procedures substantially similar to the FDA-approved, modified Karl Fischer method (Meyer and Boyd, Analytical Chem., 31:215-219, 1959; May et al., J. Biol. Standardization, 10:249-259, 1982; Centers for Biologics Evaluation and Research, FDA, Docket No. 89D-0140, 83-93; 1990) or by near infrared spectroscopy. Suitable moisture content(s) for a particular solid dressing may be determined empirically by one skilled in the art depending upon the intended application(s) thereof.
  • higher moisture contents are associated with more flexible solid dressings.
  • solid dressings intended for extremity wounds it may be preferred to have a moisture content of at least 6% and even more preferably in the range of 6% to 44%.
  • lower moisture contents are associated with more rigid solid dressings.
  • solid dressings intended for flat wounds such as wounds to the abdomen or chest, it may be preferred to have a moisture content of less than 6% and even more preferably in the range of 1% to 6%.
  • suitable moisture contents for solid dressings include, but are not limited to, the following (each value being ⁇ 0.9%): less than 53%; less than 44%; less than 28%; less than 24%; less than 16%; less than 12%; less than 6%; less than 5%; less than 4%; less than 3%; less than 2.5%; less than 2%; less than 1.4%; between 0 and 12%, non-inclusive; between 0 and 6%; between 0 and 4%; between 0 and 3%; between 0 and 2%; between 0 and 1%; between 1 and 16%; between 1 and 11%; between 1 and 8%; between 1 and 6%; between 1 and 4%; between 1 and 3%; between 1 and 2%; and between 2 and 4%.
  • the fibrinogen in the haemostatic layer(s) of the solid dressings may be any suitable fibrinogen known and available to those skilled in the art.
  • a specific fibrinogen for a particular application may be selected empirically by one skilled in the art.
  • the term “fibrinogen” is intended to include mixtures of fibrinogen and small amounts of Factor XIII/Factor XIIIa, or some other such transaminase. Such small amounts are generally recognized by those skilled in the art as usually being found in mammalian fibrinogen after it has been purified according to the methods and techniques presently known and available in the art, and typically range from 0.1 to 20 Units/mL.
  • the fibrinogen employed as the fibrinogen component of the solid dressing is a purified fibrinogen suitable for introduction into a mammal.
  • fibrinogen is a part of a mixture of human plasma proteins which include Factor XIII/XIIIa and have been purified to an appropriate level and virally inactivated.
  • a preferred aqueous solution of fibrinogen for preparation of a solid dressing contains around 37.5 mg/mL fibrinogen at a pH of around 7.4 ⁇ 0.1.
  • Suitable fibrinogen for use as the fibrinogen component has been described in the art, e.g. U.S. Pat. No. 5,716,645, and similar materials are commercially available, e.g. from sources such as Sigma-Aldrich, Enzyme Research Laboratories, Haematologic Technologies and Aniara.
  • the fibrinogen activator employed in the haemostatic layer(s) of the solid dressing may be any of the substances or mixtures of substances known by those skilled in the art to convert fibrinogen into fibrin.
  • suitable fibrinogen activators include, but are not limited to, the following: thrombins, such as human thrombin or bovine thrombin, and prothrombins, such as human prothrombin or prothrombin complex concentrate (a mixture of Factors II, VII, IX and X); snake venoms, such as batroxobin, reptilase (a mixture of batroxobin and Factor XIIIa), bothrombin, calobin, fibrozyme, and enzymes isolated from the venom of Bothrops jararacussu; and mixtures of any two or more of these.
  • thrombins such as human thrombin or bovine thrombin
  • prothrombins such as human prothrombin or prothrombin complex concentrate (a mixture of Factors II
  • the fibrinogen activator is a thrombin. More preferably, the fibrinogen activator is a mammalian thrombin, although bird and/or fish thrombin may also be employed in appropriate circumstances. While any suitable mammalian thrombin may be used in the solid dressing, the thrombin employed in the haemostatic layer is preferably a lyophilized mixture of human plasma proteins which has been sufficiently purified and virally inactivated for the intended use of the solid dressing. Suitable thrombin is available commercially from sources such as Sigma-Aldrich, Enzyme Research Laboratories, Haematologic Technologies and Biomol International.
  • a particularly preferred aqueous solution of thrombin for preparing a solid dressing contains thrombin at a potency of between 10 and 2000 ⁇ 50 International Units/mL, and more preferred at a potency of 25 ⁇ 2.5 International Units/mL.
  • Other constituents may include albumin (generally about 0.1 mg/mL) and glycine (generally about 100 mM ⁇ 0.1 mM).
  • the pH of this particularly preferred aqueous solution of thrombin is generally in the range of 6.5-7.8, and preferably 7.4 ⁇ 0.1, although a pH in the range of 5.5-8.5 may be acceptable.
  • the solid dressing may optionally further comprise one or more support layers.
  • a “support layer” refers to a material that sustains or improves the structural integrity of the solid dressing and/or the fibrin clot formed when such a dressing is applied to wounded tissue.
  • the support layer comprises a backing material on the side of the dressing opposite the side to be applied to wounded tissue.
  • a backing material may be affixed with a physiologically-acceptable adhesive or may be self-adhering (e.g. by having a sufficient surface static charge).
  • the backing material may comprise one or more resorbable materials or one or more non-resorbable materials or mixtures thereof.
  • the backing material is a single resorbable material.
  • the resorbable material may be a proteinaceous substance, such as silk, fibrin, keratin, collagen and/or gelatin.
  • the resorbable material may be a carbohydrate substance, such as alginates, chitin, cellulose, proteoglycans (e.g. poly-N-acetyl glucosamine), hyaluron and its derivatives, such as hyaluronic acid, glycolic acid polymers, lactic acid polymers, or glycolic acid/lactic acid co-polymers.
  • the resorbable material may also comprise a mixture of proteinaceous substances or a mixture of carbohydrate substances or a mixture of both proteinaceous substances and carbohydrate substances.
  • Specific resorbable material(s) may be selected empirically by those skilled in the art depending upon the intended use of the solid dressing.
  • the resorbable material is a carbohydrate substance.
  • Illustrative examples of particularly preferred resorbable materials include, but are not limited to, the materials sold under the trade names VICRYLTM (a glycolic acid/lactic acid copolymer) and DEXONTM (a glycolic acid polymer).
  • non-resorbable material Any suitable non-resorbable material known and available to those skilled in the art may be employed as the backing material.
  • suitable non-resorbable materials include, but are not limited to, plastics, silicone polymers, paper and paper products, latex, gauze and the like.
  • the support layer comprises an internal support material.
  • an internal support material is preferably fully contained within a haemostatic layer of the solid dressing, although it may be placed between two adjacent haemostatic layers in certain embodiments.
  • the internal support material may be a resorbable material or a non-resorbable material, or a mixture thereof, such as a mixture of two or more resorbable materials or a mixture of two or more non-resorbable materials or a mixture of resorbable material(s) and non-resorbable material(s).
  • the support layer may comprise a front support material on the wound-facing side of the dressing, i.e. the side to be applied to wounded tissue.
  • the front support material may be a resorbable material or a non-resorbable material, or a mixture thereof, such as a mixture of two or more resorbable materials or a mixture of two or more non-resorbable materials or a mixture of resorbable material(s) and non-resorbable material(s).
  • the solid dressing comprises both a backing material and an internal support material in addition to the haemostatic layer(s), i.e. the solid dressing comprises two support layers in addition to the haemostatic layer(s).
  • the solid dressing comprises both a front support material and an internal support material in addition to the haemostatic layer(s).
  • the solid dressing comprises a backing material, a front support material and an internal support material in addition to the haemostatic layer(s).
  • the solid dressings may also optionally further comprise a release layer in addition to the haemostatic layer(s) and support layer(s).
  • a “release layer” refers to a layer containing one or more agents (“release agents”) which promote or facilitate removal of the solid dressing from a mold in which it has been manufactured.
  • a preferred such agent is sucrose, but other suitable release agents include gelatin, mannitol, sorbitol, hyaluron and its derivatives, such as hyaluronic acid, and glucose.
  • release agents may be contained in the haemostatic layer.
  • the various layers of the inventive dressings may be affixed to one another by any suitable means known and available to those skilled in the art.
  • a physiologically-acceptable adhesive may be applied to a backing material (when present), and the haemostatic layer(s) subsequently affixed thereto.
  • the physiologically-acceptable adhesive has a shear strength and/or structure such that the backing material can be separated from the fibrin clot formed by the haemostatic layer after application of the dressing to wounded tissue. In other embodiments, the physiologically-acceptable adhesive has a shear strength and/or structure such that the backing material cannot be separated from the fibrin clot after application of the bandage to wounded tissue.
  • Suitable fibrinogens and suitable fibrinogen activators for the haemostatic layer(s) of the solid dressing may be obtained from any appropriate source known and available to those skilled in the art, including, but not limited to, the following: from commercial vendors, such as Sigma-Aldrich and Enzyme Research Laboratories; by extraction and purification from human or mammalian plasma by any of the methods known and available to those skilled in the art; from supernatants or pastes derived from plasma or 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; and/or from the fluids (e.g. blood, milk, lymph, urine or the like) of transgenic mammals (e.g. goats, sheep, cows) that contain a gene which has been introduced according to standard transgenic techniques and that expresses the desired fibrinogen and/or desired fibrinogen activator.
  • fluids e.g. blood, milk
  • the fibrinogen is a mammalian fibrinogen such as bovine fibrinogen, porcine fibrinogen, ovine fibrinogen, equine fibrinogen, caprine fibrinogen, feline fibrinogen, canine fibrinogen, murine fibrinogen or human fibrinogen.
  • the fibrinogen is bird fibrinogen or fish fibrinogen.
  • the fibrinogen may be recombinantly produced fibrinogen or transgenic fibrinogen.
  • the fibrinogen activator is a mammalian thrombin, such as bovine thrombin, porcine thrombin, ovine thrombin, equine thrombin, caprine thrombin, feline thrombin, canine thrombin, murine thrombin and human thrombin.
  • the thrombin is bird thrombin or fish thrombin.
  • the thrombin may be recombinantly produced thrombin or transgenic thrombin.
  • the purity of the fibrinogen and/or the fibrinogen activator for use in the solid dressing will be a purity known to one of ordinary skin in the relevant art to lead to the optimal efficacy and stability of the protein(s).
  • the fibrinogen and/or the fibrinogen activator has been subjected to multiple purification steps, such as precipitation, concentration, diafiltration and affinity chromatography (preferably immunoaffinity chromatography), to remove substances which cause fragmentation, activation and/or degradation of the fibrinogen and/or the fibrinogen activator during manufacture, storage and/or use of the solid dressing.
  • Illustrative examples of such substances that are preferably removed by purification include: protein contaminants, such as 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.
  • the amount of the fibrinogen activator employed in the solid dressing is preferably selected to optimize both the efficacy and stability thereof. As such, a suitable concentration for a particular application of the solid dressing may be determined empirically by one skilled in the relevant art. According to certain preferred embodiments of the present invention, when the fibrinogen activator is human thrombin, the amount of human thrombin employed is between 2.50 Units/mg of fibrinogen component and 0.025 Units/mg of the fibrinogen (all values being ⁇ 0.0009).
  • thrombin is between 0.250 Units/mg of fibrinogen and 0.062 Units/mg of fibrinogen and solid dressings wherein the amount of thrombin is between 0.125 Units/mg of fibrinogen and 0.080 Units/mg of fibrinogen.
  • the fibrinogen and the fibrinogen activator are preferably activated at the time the dressing is applied to the wounded tissue by the endogenous fluids of the patient escaping from the hemorrhaging wound.
  • the fibrinogen component and/or the thrombin may be activated by a suitable, physiologically-acceptable liquid, optionally containing any necessary co-factors and/or enzymes, prior to or during application of the dressing to the wounded tissue.
  • the haemostatic layer(s) may also contain one or more supplements, such as growth factors, drugs, polyclonal and monoclonal antibodies and other compounds.
  • supplements include, but are not limited to, the following: fibrinolysis inhibitors, such as aprotonin, tranexamic acid and epsilon-amino-caproic acid; antibiotics, such as tetracycline and ciprofloxacin, amoxicillin, and metronidazole; anticoagulants, such as activated protein C, heparin, prostacyclins, prostaglandins (particularly (PGI 2 ), 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, vasod
  • the ability of the dressings to seal an injured blood vessel was determined by an ex vivo porcine arteriotomy (EVPA) performance test, which was first described in U.S. Pat. No. 6,762,336.
  • the EVPA performance test evaluates the ability of a dressing to stop fluid flow through a hole in a porcine artery. While the procedure described in U.S. Pat. No. 6,762,336 has been shown to be useful for evaluating haemostatic dressings, it failed to replicate faithfully the requirements for success in vivo. More specifically, the procedure disclosed in U.S. Pat. No. 6,762,336 required testing at 37° C., whereas, in the real world, wounds are typically cooler than that.
  • Thrombin was formulated in CTB.
  • the final pH of the thrombin was 7.4 ⁇ 0.1.
  • the concentrations of thrombin were adjusted so that when mixed with the fibrinogen solutions as described below, the combination would produce a solution that contained 0.1 units/mg of Fibrinogen.
  • the temperature of the fibrinogen and thrombin prior to dispensing was 4° C. ⁇ 2° C. Molds were removed from the ⁇ 80° C. freezer and placed on a copper plate that was placed on top of dry ice. A repeat pipettor was filled with fibrinogen and second repeat pipettor was filled with thrombin.
  • the dressings were removed from the lyophylizer, sealed in foil pouches and stored at room temperature until testing. Subsequently, the dressings were evaluated in the EVPA, Adherence and Weight Assays.
  • Monolithic dressings were manufactured as follows: backing material was cut and placed into each PETG 2.4 ⁇ 2.4 cm mold. Twenty-five microliters of 2% sucrose was pipetted on top of each of the four corners of the backing material. Once completed the molds were placed in a ⁇ 80° C. freezer for at least 60 minutes.
  • ERL fibrinogen lot 3114 was formulated in CFB.
  • the final pH of the fibrinogen was 7.4 ⁇ 0.1.
  • the fibrinogen concentration was adjusted to 37.5 mg/ml.
  • Thrombin was formulated in CTB.
  • the final pH of the thrombin was 7.4 ⁇ 0.1.
  • the thrombin was adjusted to deliver 0.1 units/mg of Fibrinogen or 25 Units/ml thrombin. Once prepared the thrombin was placed on ice until use.
  • the temperature of the fibrinogen and thrombin prior to dispensing was 4° C. ⁇ 2° C. Molds were removed from the ⁇ 80° C.
  • Trilayer dressings were manufactured as described previously 1 , using the same materials as described above. Subsequently, the dressings were placed under conditions of 100% relative humidity at 37° C. for various times in order to increase their relative moisture content to desired levels. The dressings were evaluated visually and for their handling and other physical characteristics. Following this evaluation, a sample of each of the dressings was tested to determine their moisture content. The remaining dressings were performance tested in the EVPA, Adherence and Weight Held assays.
  • the monolithic dressings were fully functional at very high levels of moisture. As much as 28% moisture was found to retain complete functionality. When the moisture levels rose to 44%, the dressings were still functional, however some of their activity was reduced Higher levels of moisture may also retain some function.
  • the original dressings, at 2.5% moisture content, were not flexible, but had all the other desired properties including appearance, a flat surface, integrity, rapid and uncomplicated hydration and a smooth appearance post hydration. Once the moisture content was increased to 5.8%, the monolithic dressings became flexible, while retaining their functionality and desirable characteristics. They retained their flexibility, without curling or losing their integrity or appearing to form excessive amounts of fibrin prior to hydration.
  • the backing material was cut and placed into each PETG 2.4 ⁇ 2.4 cm mold. Twenty-five microliters of 2% sucrose was pipetted on top of each of the four corners of the backing material. Once completed the molds were placed in a ⁇ 80° C. freezer for at least 60 minutes. For dressings without backing material, PETG 2.4 ⁇ 2.4 cm molds were placed in a ⁇ 80° C. freezer for at least 60 minutes.
  • ERL fibrinogen lot 3114 was formulated in CFB.
  • the final pH of the fibrinogen was 7.4 ⁇ 0.1.
  • the fibrinogen concentration was adjusted to 37.5 mg/ml.
  • Thrombin was formulated in CTB.
  • the final pH of the thrombin was 7.4 ⁇ 0.1.
  • the thrombin was adjusted to deliver 0.1 units/mg of Fibrinogen or 25 Units/ml thrombin. Once prepared the thrombin was placed on ice until use.
  • the temperature of the fibrinogen and thrombin prior to dispensing was 4° C. ⁇ 2° C. Molds were removed from the ⁇ 80° C.
  • Dressings formulated with backing material performed well, with all dressings passing the EVPA test, and high values for adherence and weight held. Dressings without backing material were not quite as effective in the EVPA assay, however, surprisingly 75% of them passed the EVPA test. Without the backing the other tests could not be performed. The ability of the dressings made without a backing to succeed in the EVPA assay indicates that these dressings would be effective in treating arterial injuries and even more effective in treating venous and small vessel injuries.
  • ERL fibrinogen lot 3130 was formulated in CFB.
  • the final pH of the fibrinogen was 7.4 ⁇ 0.1.
  • the fibrinogen concentration was adjusted to 37.5 mg/ml.
  • Thrombin was formulated in CTB.
  • the final pH of the thrombin was 7.4 ⁇ 0.1.
  • the thrombin was adjusted to deliver 0.1 units/mg of Fibrinogen or 25 Units/ml thrombin.
  • this support material was cut into approximately 1 mm ⁇ 1 mm pieces and dispersed within the thrombin solution prior to filling the molds. Once prepared the thrombin was placed on ice until use.
  • the temperature of the fibrinogen and thrombin prior to dispensing was 4° C. ⁇ 2° C.
  • Cylindrical molds made of 10 or 3 mL polypropylene syringes (Becton Dickinson) with the luer-lock end removed were used. The plungers were withdrawn to the 6 mL and 2 mL mark respectively.
  • the support material was cut and placed into each mold and pushed down until it was adjacent to the plunger. Once prepared the molds were placed upright and surrounded by dry ice, leaving the opening exposed at the top.
  • both groups were performance tested in a modified EVPA assay. Briefly, a plastic foam form was slipped over the artery. This covering had a hole in it that corresponded to the hole in the artery and the surrounding tissue. Warm saline was added to the surface of the dressing and the mold was immediately passed down thru the hole in the foam to the artery surface. The plunger was then depressed and held by hand for 3 minutes, after which the mold was withdrawn as the plunger was depressed further. At this point the artery was pressurized and the assay continued as before.
  • Dressings that included no backing or a DEXONTM mesh backing performed well, with all passing the EVPA test at 250 mmHg.
  • the dressings also performed well, with the large size (10 mL mold) dressings holding the full 250 mmHg of pressure, while the smaller held up to 150 mmHg of pressure. This indicates that the use of a support material may be optional, and it's location may be on the ‘back’ of the dressing, or dispersed thou the composition, as desired.
  • Dressings made with a support material on the “back” (i.e. the non wound-facing side) of the dressing were manufactured by first cutting the mesh support material and placing it into each PETG 10 ⁇ 10 cm mold. Twenty-five microliters of 2% sucrose was pipetted on top of each of the four corners of the backing material. Once completed the molds were placed in a ⁇ 80° C. freezer for at least 60 minutes.
  • ERL fibrinogen lot 3114 was formulated in CFB.
  • the final pH of the fibrinogen was 7.4 ⁇ 0.1.
  • the fibrinogen concentration was adjusted to 37.5 mg/ml.
  • Thrombin was formulated in CTB.
  • the final pH of the thrombin was 7.4 ⁇ 0.1.
  • the thrombin was adjusted to deliver 0.1 units/mg of Fibrinogen or 25 Units/ml thrombin. Once prepared the thrombin was placed on ice until use.
  • the thrombin was adjusted to deliver 0.1 units/mg of Fibrinogen or 25 Units/ml thrombin. Once prepared the thrombin was placed on ice until use.
  • the temperature of the fibrinogen and thrombin prior to dispensing was 4° C. ⁇ 2° C.
  • the mold was removed from the ⁇ 80° C. freezer and placed on an aluminum plate that was placed on top of dry ice.
  • the aluminum plate had a 0.25 inch hole drilled in the center and a fitting attached so that a piece of tubing could be attached to a vacuum source.
  • the mold was centered over the hole in the aluminum plate and vacuum was turned on. The vacuum served two purposes it prevented the mold from moving and it held it flat against the aluminum plate.
  • Thirty-five milliliters of fibrinogen and 5.25 milliliters of Thrombin were placed in 50 ml test tube, inverted three times and poured into the mold. Once the molds were filled and the support material applied as described above, they were returned to the ⁇ 80° C. freezer for at least two hours before being placed into the freeze dryer. Dressings were then lyophylized as described previously.
  • EXCLUSION CRITERION The mesh backing must remain over the hole in the artery. If it has shifted during the polymerization and does not completely cover the hole the haemostatic dressing must be excluded.
  • the set-up of the testing equipment is shown in FIG. 2 .
  • Some additional, unshown components may be utilized to read out (pressure gauge) or control the pressure within the system.
  • the pressure/time profile to be followed is attained by manually turning the pump on and off while referencing the system pressure as read out by one or more pressure-reading components of the system.
  • the haemostatic dressing is subjectively assessed with regard to adhesion to the artery and formation of a plug in the artery hole. Any variations from the positive control should be noted on the data collection form.
  • Haemostatic dressings that are able to withstand pressures for 3 minutes are considered to have passed the assay.
  • the data collection should be stopped immediately so that the natural decrease in pressure that occurs in the artery once the test is ended isn't included on the graphs. Should the operator fail to stop data collection, these points can be deleted from the data file to avoid confusing the natural pressure decay that occurs post-test with an actual dressing failure. The entire testing period from application of the haemostatic dressing to completion must fall within pre-established criteria. The maximum pressure reached should be recorded on the data collection form.
  • Haemostatic dressings that start leaking saline at any point during testing are considered to have failed the assay.
  • the pressure should be allowed to fall ⁇ 20 mmHg before data collection is stopped so that the failure is easily observed on the graphs.
  • the pressures at which leakage occurred should be recorded on the data collection form. Should the data collection stop in the middle of the experiment due to equipment failure the data can be collected by hand at 5 second intervals until the end of the test or haemostatic dressing failure, whichever happens first.
  • the data points should be recorded on the back of the data collection form, clearly labeled, and entered by hand into the data tables.
  • results must be excluded. If there are leaks from collaterals that can't be fixed either by patching or finger pressure the results must be excluded. If the test fails because of leaks at the O-rings, the results must be excluded. If the mesh backing does not completely cover the hole in the artery, the results must be excluded.
  • Hemostat(s), Porcine artery and haemostatic dressing (usually after completion of the EVPA Assay although it does not need to be performed to do the Adherence Assay).
  • the dressing After application of the dressing without completion of the EVPA Assay, the dressing is ready for the Adherence Assay and Weight Limit Test (if applicable). After application of the dressing and subsequent EVPA Analysis, the artery and syringe system is then disconnected slowly from the pump so that solution does not spray everywhere. The warmed, red saline solution from the EVPA Assay remains in the syringe until the Adherence Assay and Weight Limit Test (if applicable) is completed.
  • the mesh backing must remain over the hole in the artery. If it has shifted during the polymerization and does not completely cover the hole the haemostatic dressing must be excluded.
  • a dressing does not adhere to the artery after application and/or prior to performing the EVPA assay, it is given a score of 0 and fails the adherence test. If a dressing receives a score ⁇ 2, the dressing is considered to have failed the Adherence Assay.
  • weights may then be added to the hemostat in an incremental manner until the mesh backing is pulled entirely off of the artery.
  • the maximum weight that the dressing holds is then recorded as a measure of the amount of weight the dressing could hold attached to the artery.
  • Moisture determinations were carried out using a Brinkman Metrohm Moisture Analyzer System.
  • the system contains the following individual components, 774 Oven Sample Processor, 774SC Controller, 836 Titrando, 5 ml and 50 ml 800 Dosino Units and a 801 Stirrer.
  • the system was connected to a computer using the Brinkman Tiamo software for data collection, analysis and storage.
  • the moisture system is set-up and run according to the manufactures recommendations and specifications to measure the moisture content of lyophilized samples using the Karl Fischer method.

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US11/882,876 2006-08-04 2007-08-06 Solid dressing for treating wounded tissue Abandoned US20080033332A1 (en)

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US11/882,876 US20080033332A1 (en) 2006-08-04 2007-08-06 Solid dressing for treating wounded tissue
US13/363,489 US20120148658A1 (en) 2006-08-04 2012-02-01 Solid dressing for treating wounded tissue
US14/599,519 US20150132363A1 (en) 2006-08-04 2015-01-18 Solid dressing for treating wounded tissue
US15/208,591 US20160317355A1 (en) 2006-08-04 2016-07-12 Solid dressing for treating wounded tissue
US15/605,660 US20180117210A1 (en) 2006-08-04 2017-05-25 Solid dressing for treating wounded tissue and processes for mixing fibrinogen and thrombin while preserving fibrin-forming ability, compositions produced by these processes, and the use thereof
US16/289,535 US20200000957A1 (en) 2006-08-04 2019-02-28 Solid dressing for treating wounded tissue and processes for mixing fibrinogen and thrombin while preserving fibrin-forming ability, compositions produced by these processes, and the use thereof

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US11/882,876 US20080033332A1 (en) 2006-08-04 2007-08-06 Solid dressing for treating wounded tissue

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US11/882,874 Abandoned US20080033331A1 (en) 2006-08-04 2007-08-06 Solid dressing for treating wounded tissue
US11/882,879 Abandoned US20080033333A1 (en) 2006-08-04 2007-08-06 Solid dressing for treating wounded tissue
US13/363,489 Abandoned US20120148658A1 (en) 2006-08-04 2012-02-01 Solid dressing for treating wounded tissue
US13/364,762 Abandoned US20120148659A1 (en) 2006-08-04 2012-02-02 Solid dressing for treating wounded tissue
US13/364,837 Abandoned US20120150087A1 (en) 2006-08-04 2012-02-02 Solid dressing for treating wounded tissue
US14/583,002 Abandoned US20150118284A1 (en) 2006-08-04 2014-12-24 Solid dressing for treating wounded tissue
US14/599,519 Abandoned US20150132363A1 (en) 2006-08-04 2015-01-18 Solid dressing for treating wounded tissue
US14/746,482 Abandoned US20150283288A1 (en) 2006-08-04 2015-06-22 Solid dressing for treating wounded tissue
US15/088,438 Abandoned US20160213806A1 (en) 2006-08-04 2016-04-01 Solid dressing for treating wounded tissue
US15/208,563 Abandoned US20160317700A1 (en) 2006-08-04 2016-07-12 Solid dressing for treating wounded tissue
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US11/882,874 Abandoned US20080033331A1 (en) 2006-08-04 2007-08-06 Solid dressing for treating wounded tissue
US11/882,879 Abandoned US20080033333A1 (en) 2006-08-04 2007-08-06 Solid dressing for treating wounded tissue
US13/363,489 Abandoned US20120148658A1 (en) 2006-08-04 2012-02-01 Solid dressing for treating wounded tissue
US13/364,762 Abandoned US20120148659A1 (en) 2006-08-04 2012-02-02 Solid dressing for treating wounded tissue
US13/364,837 Abandoned US20120150087A1 (en) 2006-08-04 2012-02-02 Solid dressing for treating wounded tissue
US14/583,002 Abandoned US20150118284A1 (en) 2006-08-04 2014-12-24 Solid dressing for treating wounded tissue
US14/599,519 Abandoned US20150132363A1 (en) 2006-08-04 2015-01-18 Solid dressing for treating wounded tissue
US14/746,482 Abandoned US20150283288A1 (en) 2006-08-04 2015-06-22 Solid dressing for treating wounded tissue
US15/088,438 Abandoned US20160213806A1 (en) 2006-08-04 2016-04-01 Solid dressing for treating wounded tissue
US15/208,563 Abandoned US20160317700A1 (en) 2006-08-04 2016-07-12 Solid dressing for treating wounded tissue
US15/208,591 Abandoned US20160317355A1 (en) 2006-08-04 2016-07-12 Solid dressing for treating wounded tissue

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