WO2017223462A1 - Composition et procédé pour coller des biomatériaux à une surface cible - Google Patents

Composition et procédé pour coller des biomatériaux à une surface cible Download PDF

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Publication number
WO2017223462A1
WO2017223462A1 PCT/US2017/039019 US2017039019W WO2017223462A1 WO 2017223462 A1 WO2017223462 A1 WO 2017223462A1 US 2017039019 W US2017039019 W US 2017039019W WO 2017223462 A1 WO2017223462 A1 WO 2017223462A1
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Prior art keywords
transglutaminase
composition
glutamic acid
wounds
polymer
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PCT/US2017/039019
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English (en)
Inventor
Donald E. Ingber
Javier G. FERNANDEZ
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President And Fellows Of Harvard College
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Publication of WO2017223462A1 publication Critical patent/WO2017223462A1/fr

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    • 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
    • A61L26/00—Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form
    • A61L26/0061—Use of materials characterised by their function or physical properties
    • A61L26/0066—Medicaments; Biocides
    • 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
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/1703—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
    • A61K38/1709—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
    • 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
    • A61K38/45—Transferases (2)
    • 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
    • A61L24/00—Surgical adhesives or cements; Adhesives for colostomy devices
    • A61L24/001—Use of materials characterised by their function or physical properties
    • A61L24/0015—Medicaments; Biocides
    • 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
    • A61L24/00—Surgical adhesives or cements; Adhesives for colostomy devices
    • A61L24/04—Surgical adhesives or cements; Adhesives for colostomy devices containing macromolecular materials
    • A61L24/10—Polypeptides; Proteins
    • A61L24/108—Specific proteins or polypeptides not covered by groups A61L24/102 - A61L24/106
    • 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
    • A61L26/00—Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form
    • A61L26/0009—Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form containing macromolecular materials
    • A61L26/0028—Polypeptides; Proteins; Degradation products thereof
    • A61L26/0047—Specific proteins or polypeptides not covered by groups A61L26/0033 - A61L26/0042
    • 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
    • A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
    • A61L2300/20—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices containing or releasing organic materials
    • A61L2300/252—Polypeptides, proteins, e.g. glycoproteins, lipoproteins, cytokines
    • A61L2300/254—Enzymes, proenzymes
    • 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
    • A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
    • 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
    • A61L2300/412—Tissue-regenerating or healing or proliferative agents

Definitions

  • the present invention is directed to compositions and method for bonding biomatenals formed from carbohydrates and proteins to organic and inorganic surfaces. More specifically, the invention relates to compositions comprising transglutaminase (TG) and glutamic acid rich polypeptides, such as casein, and their use for bonding biomaterials, such as chitosan and collagen biomaterials, to surfaces of organic and inorganic substrates. The invention also relates to compositions and methods for wound healing.
  • TG transglutaminase
  • glutamic acid rich polypeptides such as casein
  • biomaterials such as chitosan and collagen biomaterials
  • Natural biomaterials such as chitosan and collagen
  • compositions and methods that can rapidly and tightly bond biomaterials to surface of organic (such as living tissue) and inorganic substrates (such as materials used in medical products). The present invention partly addresses this need.
  • the invention provides a method for bonding or adhering a biomaterial to a target surface.
  • the method comprises applying an effective amount of a transglutaminase and a glutamic acid rich polypeptide to a target surface and contacting a biomaterial to the target surface where the transglutaminase and the glutamic acid rich polypeptide have been applied.
  • the invention provides a method for promoting wound healing.
  • the method comprises applying a transglutaminase and a glutamic acid rich polypeptide to a surface of a wound.
  • the invention provides a method for forming a coating layer on a target surface.
  • the method comprises applying a polymer, a transglutaminase and a glutamic acid rich polypeptide to the same portion of a target surface.
  • the coating layer can act as an adhesive, e.g., as a dressing on a wound surface, or as a glue to bond two surfaces together.
  • the adhesive coating layer is selective for adhesion to the target surface.
  • the coating layer can act as a sealant or provide a physical barrier, e.g., to reduce or stop a fluid leakage or permeation such as bleeding, or to protect a substrate or surface being coated from an external disturbance (e.g., bacteria, light, moisture, or any combinations thereof).
  • an external disturbance e.g., bacteria, light, moisture, or any combinations thereof.
  • the coating layer can form a casing, e.g., a "skin" to enclose a material therein.
  • the coating layer can seal lung punctures.
  • the invention provides a three component composition.
  • the composition comprises as a first component a polymer in a flowable form, wherein the polymer comprises an amino group; as a separate second component a transglutaminase in a flowable form; and as a separate third component a glutamic acid rich polypeptide in a flowable form.
  • the glutamic acid rich polypeptide is casein.
  • the transglutaminase is a mammalian or microbial transglutaminase.
  • FIGS. la-Id show the microbial transglutaminase (mTG) reaction and mTG- mediated bonding of biomaterial films.
  • FIG. la shows mTG-catalyzed reactions with relevance for biomaterial bonding.
  • the upper reaction is a deamination that occurs in the absence of amine substrates and the presence of water.
  • mTG catalyzes the hydrolysis of the glutaminyl residue, resulting in loss of the amine group.
  • the presence of a primary amine group results in formation of a covalent bond between both molecules.
  • the involved atoms are O (red), N (blue) and H (white).
  • FIG. lb is a bar graph showing bonding strengths produced by commercial mTG preparations containing mTG and maltodextrin (mTG+Ma) or both
  • FIG. lc is a graph showing kinetics of the mTG bonding reaction measured at the macroscale as the adhesive force between surfaces of two muscle tissues.
  • FIG. Id is a graph showing more detailed examination of the first 15 minutes of the reaction dynamics shown in FIG. lc.
  • FIGS. 2a-2b show mTG-mediated bonding of biomaterials to an inorganic PDMS substrate.
  • FIG. 2a shows the chemistry that underlies APTES functionalization of the PDMS surface.
  • the highly reactive silane group in APTES silanizes the surface by forming covalent bonds with surface atoms.
  • FIG. 2b is a bar graph showing bonding strength produced between contacting chitosan or collagen films and untreated (PDMS) or APTES-modified PDMS
  • FIGS. 3a-3e show the applicability of mTG bonding of biomaterial films and foams to native tissues.
  • FIG. 3a shows the diagram of the ASTM F2392 standard protocol to measure burst strength of surgical sealants that are used to fill a 3 mm diameter hole on a collagen surface.
  • FIG. 3b is a bar graph showing performance of different sealing materials tested in the ASTM F2392 assay presented as average maximum and ultimate pressures required to burst the seals.
  • FIG. 3c is a bar graph showing bonding strength of a chitosan film bonded to heart and liver surface using mTG and casein (mTG+Ca) preparation.
  • FIG. 3d is a bar graph showing bonding strength of a chitosan film adhered to skin epidermis or dermis (after epidermis is removed); control indicates bonding of chitosan to dermis, however no adhesion is observed to undamaged (fully cornified) epidermis.
  • FIG. 3e shows a photomicrograph of a chitosan foam bonded to the surfaces of a 1 cm irregularly shaped defect in an explanted pig muscle using mTG+Ca (note the seamless nature of the adhesion).
  • FIGS. 4a-4e show use of mTG as a sealant for tissue punctures.
  • FIG. 4a is a graph showing the burst test performed on a 1 cm diameter hole in the wall of a small intestine on an explanted pig, repaired with a chitosan film patch bonded to the tissue with mTG+Ca. The patch resisted a linear increment of hydraulic pressure applied through the intestinal lumen for several minutes, reaching the equivalent to the maximum physiological pressures in the human body before bursting.
  • FIG. 4b shows images of the chitosan patch before (top) and after (bottom) on a region of the native intestine on the opposite side from where the patch burst at high pressure, corresponding to point C on the graph shown in FIG.
  • FIG. 4c shows image and digital rendering of the double-canister spray device designed to form a conformal coating of chitosan with mTG+Ca with adhesive properties. Each component is loaded into separate barrels, and pushed through the nozzles using the same pressure source.
  • FIG. 4d shows photographs of a 3 cm deep puncture in an explanted pig lung before (top) and after (bottom) it was sprayed for 10 min while cyclically inflating and deflating the lungs using the double-canister spray method (data not shown).
  • FIG. 4e shows a scanning electron microscopic image of the
  • Embodiments of the various aspects disclosed herein include a transglutaminase.
  • Embodiments of the various aspect disclosed herein include applying different components, such as one or more of a biomaterial, a transglutaminase, a glutamic acid rich polypeptide and a polymer to a target surface.
  • the various components that are applied to the target surface can be formulated in a solution, an emulsion, an aerosol, a foam, an ointment, a paste, a lotion, a powder, a gel, a hydrogel, a hydrocolloid, a microparticle, a nanoparticle, or a cream.
  • the components can be formulated in separate compositions, all together in one composition, or some together in one composition and others in separate compositions.
  • the transglutaminase and the glutamic acid rich polypeptide are formulated in one composition.
  • the biomaterial or the polymer is formulated in a separate composition and the transglutaminase and the glutamic acid rich polypeptide are formulated in one composition.
  • At least one of the biomaterial, transglutaminase and glutamic acid rich polypeptide or at least one of the polymer, transglutaminase and the glutamic acid rich polyopeptide can be provided in a flowable form, e.g., a state of a material that is capable of flowing.
  • Examples of a flowable form include, but are not limited to, a liquid, a fluid, a solution, powder, particles (e.g., nanoparticles, or microparticles), or fibers (e.g., nanofibers or microfibers), a suspension, a colloid, a gel, an emulsion (e.g., oil-in-water or water-in-oil), an aerosol, a foam, an ointment, a paste, or any combinations thereof.
  • at least two of the biomaterial, transglutaminase, glutamic acid rich polypeptide and polymer are in liquid form.
  • at least two of the biomaterial, transglutaminase, glutamic acid rich polypeptide and polymer are in powder form.
  • Selection of an appropriate flowable form for the biomaterial, transglutaminase, glutamic acid rich polypeptide and polymer can be determined based on a number of factors, e.g., but not limited to, viscosity and/or solubility of the biomaterial, transglutaminase, glutamic acid rich polypeptide and/or polymer, reaction rate (and reaction time) of crosslinking upon contact, and any combinations thereof.
  • the biomaterial or polymer used in the compositions and methods described herein comprises chitosan
  • it is more desirable to provide chitosan in a liquid than in powder e.g., pre-dissolution of chitosan in a solvent vs. chitosan powder
  • chitosan generally requires a specific pH condition for dissolution, and its dissolution rate is much slower than a crosslinking reaction rate with the transglutaminase (e.g., on a time scale of less than a second).
  • chitosan were provided in powder for use in the methods described herein, contacting chitosan powder in a transglutaminase solution could potentially result in formation of a shell of cross-linked chitosan encapsulating unreacted chitosan. Accordingly, pre-dissolution of chitosan in an acidic solvent can be more beneficial for formation of a more homogenous coating layer. Similarly, in some embodiments, it is also more desirable to provide a solution of transglutaminase so that it can react with a chitosan solution to form a more homogenous coating layer upon contact on a target surface.
  • the biomaterial, transglutaminase, glutamic acid rich polypeptide and polymer in a flowable form can be applied to a target surface by any methods known in the art to deliver a flowable material, including, but not limited to, extrusion, atomization, spraying, pumping, and any combinations thereof.
  • application to a target surface is by a delivery method comprising atomization of the component to be applied.
  • Exemplary atomization methods can include, but are not limited to, syringe extrusion, coaxial air flow method, mechanical disturbance method, electrostatic force method, electrostatic bead generator method, spraying, atomization using a rotary or centrifugal atomizer, air atomization (e.g., using a spray gun and air pressure), pressure atomization, vacuum atomization (e.g., by spraying from high pressure into low pressure zone), ultrasonic atomization, and any combinations thereof.
  • solutions can be broken into fine droplets with the aid of air flow pressure.
  • the air flow pattern can be altered to form coaxial pattern for formation of uniform droplets or particles.
  • Coaxial air flow technique generally uses concentric streams of air which shear the liquid droplets released from one or more needles.
  • Alternatives to the air driven mechanism include electrostatic field, mechanical disturbance and electrostatic force.
  • Electrostatic mechanism generally utilizes a potential difference between a capillary tip such as a nozzle and a flat counter electrode to reduce the diameter of the droplets by applying an additional force (i.e., electric force) in the direction of gravitational force in order to overcome the upward capillary force of liquid.
  • mechanical disturbance method liquid droplets can be broken into fine droplets using a mechanical disturbance.
  • vibrations including ultrasonic atomization can be as a mechanical disturbance to produce fine droplets.
  • electrostatic forces can destabilize a viscous jet, where the electrostatic force can be used to disrupt the liquid surface instead of a mechanical disturbance.
  • each atomization condition can be independently controlled to provide a desired atomized droplet size.
  • an aerosol of a solution can be controlled by changing instrumental/process, and/or material parameters.
  • Exemplary instrumental/process parameters that can be varied include, but are not limited to, air pressure of a spray, nozzle size (e.g., nozzle diameter), atomization power output, flow rate of a spray, height of a nozzle head (e.g., distance of the nozzle head from a target surface), atomization duration, and material parameters that can be varied include, but are not limited to, concentration and/or viscosity of the solution, and/or concentration of a plasticizer, if any.
  • biomaterial, transglutaminase, glutamic acid rich polypeptide or the polymer, transglutaminase and the glutamic acid rich polypeptide can be sprayed to a target surface, e.g., forming an aerosol or a mist of fine particles.
  • the delivery method can further comprise mixing the amino-polymer and transglutaminase. For example, when the amino-polymer and the transglutaminase are to be sprayed to a target surface, each component can be sprayed at an angle such that both components contact with each other in air and a mixing of the components occurs during the flight in air.
  • the biomaterial or polymer, transglutaminase and glutamic acid rich polypeptide can be applied to the same portion of a target surface in a sequential or concurrent manner.
  • the components can be applied concurrently or simultaneously to the same portion of a target surface.
  • the components can be individually applied to a target surface at the same time such that both components can contact and mix with each other before the mixture deposits on the target surface.
  • the components can be sprayed as separate entities concurrently or simultaneously to the same portion of a target surface such that the sprayed components contact and mix with each other during flight in air prior to depositing on the target surface.
  • the components can be applied as a single mixture to a target surface, provided that no significant pre-crosslinking in the mixture of the components occurs prior to its application to the target surface.
  • the crosslinking reaction is generally rapid (e.g., on a time scale of less than a second)
  • the pre-determined volume ratio of different components applied to a target substrate or surface can vary with a number of factors, including, but not limited to, selection of an appropriate biomaterial or polymer, viscosity of the different compositions to be used, desired properties of a resulting material.
  • the pre-determined volume ratio of two components to be applied can range from about 10: 1 to about 1 : 10, or from about 5: 1 to about 1 :5, or from about 3 : 1 to about 1 :3.
  • the pre-determined volume ratio can range from about 1 : 1 to about 1 :5, or from about 1 : 1 to about 1 : 3.
  • the method can further comprise applying an additive, in addition to the biomaterial or polymer and the transglutaminase and glutamic acid rich polypeptide, to a target surface.
  • the additive can be mixed with one of the other prior to applying to a target surface.
  • the additive can be independently applied to a target surface in concurrent with the application of the other components to the target surface.
  • An additive can be any molecule, compound, or agent that can produce an effect (e.g., a beneficial effect) on and/or in close proximity to the target surface, and/or that can confer a new property/feature to the target surface.
  • an additive can include, without limitations, a living cell (e.g., a stem cell), a therapeutic agent, a hemostatic agent, an antiseptic agent (e.g., an antibiotic), a wound healing agent, a cross-linking agent, flavorings, colorings, nutraceuticals, a cell growth factor, a peptide, a peptidomimetic, an antibody or a portion thereof, an antibody-like molecule, nucleic acid, a plasticizer (e.g., glycerol), a nanoparticle or microparticle, a nanofiber, and any combinations thereof.
  • a living cell e.g., a stem cell
  • an antiseptic agent e.g., an antibiotic
  • a wound healing agent e.g., a wound healing agent
  • a cross-linking agent e.g., flavorings, colorings, nutraceuticals, a cell growth factor, a peptide, a peptidomimetic, an antibody or a portion
  • the methods disclosed herein can be performed on a target substrate or surface more than once (e.g., twice, three times, four times or more).
  • a second polymer composition and a second transglutaminase composition can be applied on the first coating layer, thereby forming a second coating layer on top of the coating layer.
  • the applied second amino- polymer composition (including, e.g., types and/or concentrations of the polymer(s), and optionally additive(s)) can be the same as or different from the first polymer composition.
  • the second transglutaminase composition (including, e.g., sources and/or origins of transglutaminases) can be the same as or different from the first transglutaminase composition. Accordingly, a method for providing a multi-layer coating layer is also provided herein.
  • the biomaterial, transglutaminase, glutamic acid rich polypeptide and polymer can be provided in an aerosol.
  • at least one (e.g., one, two, three or all four) of the biomaterial, transglutaminase, glutamic acid rich polypeptide and polymer is an aerosol.
  • a target surface can be any surface to which a biomaterial or a polymer is to be bonded or adhered.
  • a target surface can be surface of an organic or inorganic substrate.
  • the target surface can be surface of a tissue or organ.
  • the target surface can include surfaces of hepatic, cardiac and dermal tissues.
  • target surface includes a wound.
  • wound refers to physical disruption of the continuity or integrity of tissue structure caused by a physical (e.g., mechanical) force, a biological (e.g., thermic or actinic force), or a chemical means.
  • the term “wound” encompasses wounds of the skin.
  • wound also encompasses contused wounds, as well as incised, stab, lacerated, open, penetrating, puncture, abrasions, grazes, burns, frostbites, corrosions, wounds caused by ripping, scratching, pressure, and biting, and other types of wounds.
  • wound also includes surgical wounds.
  • the wound can be acute or chronic.
  • chronic wound refers to a wound that does not fully heal even after a prolonged period of time (e.g., 2 to 3 months or longer).
  • Chronic wounds including pressure sores, venous leg ulcers and diabetic foot ulcers, can simply be described as wounds that fail to heal. Whilst the exact molecular pathogenesis of chronic wounds is not fully understood, it is acknowledged to be multi-factorial. As the normal responses of resident and migratory cells during acute injury become impaired, these wounds are characterized by a prolonged inflammatory response, defective wound extracellular matrix (ECM) remodelling and a failure of re-epithelialisation.
  • ECM defective wound extracellular matrix
  • the wound can be an internal wound, e.g. where the external structural integrity of the skin is maintained, such as in bruising or internal ulceration, or external wounds, particularly cutaneous wounds, and consequently the tissue can be any internal or external bodily tissue.
  • the tissue is skin (such as human skin), i.e. the wound is a cutaneous wound, such as a dermal or epidermal wound.
  • Wounds can be classified in one of two general categories, partial thickness wounds or full thickness wounds.
  • a partial thickness wound is limited to the epidermis and superficial dermis with no damage to the dermal blood vessels.
  • a full thickness wound involves disruption of the dermis and extends to deeper tissue layers, involving disruption of the dermal blood vessels.
  • the healing of the partial thickness wound occurs by simple regeneration of epithelial tissue. Wound healing in full thickness wounds is more complex.
  • the wound is selected from the group consisting of cuts and lacerations, surgical incisions or wounds, punctures, grazes, scratches, compression wounds, abrasions, friction wounds (e.g. nappy rash, friction blisters), decubitus ulcers (e.g. pressure or bed sores); thermal effect wounds (burns from cold and heat sources, either directly or through conduction, convection, or radiation, and electrical sources), chemical wounds (e.g. acid or alkali burns) or pathogenic infections (e.g.
  • viral, bacterial or fungal including open or intact boils, skin eruptions, blemishes and acne, ulcers, chronic wounds, (including diabetic-associated wounds such as lower leg and foot ulcers, venous leg ulcers and pressure sores), skin graft/transplant donor and recipient sites, immune response conditions, e.g. psoriasis and eczema, stomach or intestinal ulcers, oral wounds, including a ulcers of the mouth, damaged cartilage or bone, amputation wounds, corneal lesions, and any combinations thereof.
  • the wound can be selected from the group consisting of cuts and lacerations, surgical incisions, punctures, grazes, scratches, compression wounds, abrasions, friction wounds, chronic wounds, ulcers, thermal effect wounds, chemical wounds, wounds resulting from pathogenic infections, skin graft/transplant donor and recipient sites, immune response conditions, oral wounds, stomach or intestinal wounds, damaged cartilage or bone, amputation sites, corneal lesions and lung punctures.
  • compositions and methods disclosed herein can be used for wound healing.
  • wound healing refers to a regenerative process with the induction of an exact temporal and spatial healing program comprising wound closure and the processes involved in wound closure.
  • wound healing encompasses but is not limited to the processes of granulation, neovascularization, fibroblast, endothelial and epithelial cell migration, extracellular matrix deposition, reepithelialization, and remodeling.
  • wound healing includes the restoration of tissue integrity. It will be understood that this can refer to a partial or a full restoration of tissue integrity. Treatment of a wound thus refers to the promotion, improvement, progression, acceleration, or otherwise advancement of one or more stages or processes associated with the wound healing process.
  • wound closure refers to the healing of a wound wherein sides of the wound are rejoined to form a continuous barrier (e.g., intact skin).
  • granulation refers to the process whereby small, red, grain-like prominences form on a raw surface (that of wounds) as healing agents.
  • neovascularization refers to the new growth of blood vessels with the result that the oxygen and nutrient supply is improved.
  • angiogenesis refers to the vascularization process involving the development of new capillary blood vessels.
  • cell migration refers to the movement of cells (e.g., fibroblast, endothelial, epithelial, etc.) to the wound site.
  • extracellular matrix deposition refers to the secretion by cells of fibrous elements (e.g., collagen, elastin, reticulin), link proteins (e.g., fibronectin, laminin), and space filling molecules (e.g., glycosaminoglycans).
  • fibrous elements e.g., collagen, elastin, reticulin
  • link proteins e.g., fibronectin, laminin
  • space filling molecules e.g., glycosaminoglycans.
  • reepithelialization refers to the reformation of epithelium over a denuded surface (e.g., wound).
  • reconstructing refers to the replacement of and/or devascularization of granulation tissue.
  • a wound healing agent can be applied to the wound.
  • a wound healing agent is a compound or composition that actively promotes wound healing process.
  • exemplary wound healing agents include, but are not limited to dexpanthenol; growth factors; enzymes, hormones; povidon-iodide; fatty acids; anti-inflammatory agents; antibiotics; antimicrobials; antiseptics; cytokines; thrombin; angalgesics; opioids; aminoxyls; furoxans; nitrosothiols; nitrates and anthocyanins; nucleosides, such as adenosine; and nucleotides, such as adenosine diphosphate (ADP) and adenosine triphosphate (ATP); neutotransmitter/neuromodulators, such as acetylcholine and 5- hydroxytryptamine (serotonin/5 -HT).
  • ADP adenosine diphosphate
  • ATP
  • Exemplary growth factors include, but are not limited to, fibroblast growth factor (FGF), FGF-1, FGF-2, FGF-4, FGF-a, FGF- ⁇ , plateletderived growth factor (PDGF), insulin- binding growth factor (IGF), IGF-1, IGF-2, heparin-binding growth factor- 1, heparin-binding growth factor-2, epidermal growth factor (EGF), transforming growth factor (TGF), TGF-a, TGF- ⁇ , cartilage inducing factors-A and -B, osteoid-inducing factors, osteogenin, vascular endothelial growth factor, bone growth factors, collagen growth factors, insulin-like growth factors, and their biologically active derivatives.
  • FGF fibroblast growth factor
  • FGF-1 FGF-1, FGF-2, FGF-4, FGF-a, FGF- ⁇
  • PDGF plateletderived growth factor
  • IGF insulin- binding growth factor
  • IGF insulin- binding growth factor
  • IGF insulin-bind growth factor
  • IGF insulin
  • transglutaminase is meant a member of the group of enzymes identified by Enzyme Commission System of Classification No. 2.3.2.13 (EC 2.3.2.13). Skilled artisan is well aware that transglutaminases are enzymes that catalyze an acyl transfer reaction of a ⁇ - carboxamide group of a glutamine residue in a peptide chain. Transglutaminases form 8-(y-Glu)- Lys crosslinks in and between protein molecules when an ⁇ -amino group of a lysine residue in a protein acts as an acyl receptor. Transglutaminases can also deaminate a glutamine residue into a glutamic acid residue when water acts as an acyl receptor.
  • transglutaminases include calcium-independent transglutaminases and calcium- dependent transglutaminases.
  • the former include an enzyme derived from microorganisms (see, for example, JP-A-1-27471, content of which is incorporated herein by reference in its entirety), and the latter include an enzyme derived from the guinea pig's liver (see, JP-B-150382, content of which is incorporated herein by reference in its entirety), an enzyme derived from fish (see, for example, Seki Nobuo et al. Nihon Suisan Gakkaishi, vol. 56, No. 1, p. 125 (1990), content of which is incorporated herein in its entirety), and the like. Further, it includes enzymes produced by gene recombination. See JP-A-1-300889, JP-A-5-199883, and JP-A-6-225775, contents of all which are incorporated herein by reference in their entireties).
  • the transglutaminase for use in the methods of the present invention can be from a natural or a synthetic source, e.g., recombinant.
  • transglutaminase prepared (i.e. extracted) from mammalian tissue samples, as well as mammalian transglutaminases expressed by recombinant means are included herein.
  • variants of naturally-occurring mammalian transglutaminases are also included.
  • the transglutaminase is a mammalian transglutaminase.
  • Mammalian transglutaminases can be obtained from animal cells and tissues and cellular products.
  • transglutaminase is a tissue transglutaminase (tTgase).
  • transglutaminase is a microbial transglutaminase.
  • a microbial transglutaminase can be isolated from one or more of a Streptomyces hygroscopicus strain, Streptoverticillium Baldaccii, Streptoverticillium mobaraense, or Escherichia Coli. See for example, Cui L et al., Bioresource Technology (2008) 99(9): 3794-3800, content of which is incorporated herein by reference in its entirety).
  • transglutaminase is a human transglutaminase.
  • Human transglutaminase can be prepared from human tissue or cells.
  • a human transglutaminase can be extracted from human tissue sources such as lung, liver, spleen, kidney, heart muscle, skeletal muscle, eye lens, endothelial cells, erythrocytes, smooth muscle cells, bone and macrophages.
  • human transglutaminase can be obtained from a culture of human cells that express a mammalian transglutaminase, using cell culture methodology well known in the art.
  • Preferred cell line sources of such transglutaminases include, but are not limited to, human endothelial cell line ECV304 (for tissue transglutaminase) and human osteosarcoma cell line MG63.
  • transglutaminase is a calcium-independent transglutaminase.
  • transglutaminase is a calcium-dependent transglutaminase.
  • transglutaminases include, but are not limited to, Factor XIII A (fibrin stabilizing factor), Type 1 transglutaminase (keratinocyte transglutaminase,), Type 2 transglutaminase (tissue transglutaminase), Type 3 transglutaminase (epidermal transglutaminase), Type 4 transglutaminase (prostate transglutaminase), Type 5 transglutaminase (Transglutaminase X), Type 6 transglutaminase (Transglutaminase Y), and Type 7 transglutaminase (Transglutaminase Z).
  • Factor XIII A fibrin stabilizing factor
  • Type 1 transglutaminase keratinocyte transglutaminase
  • Type 2 transglutaminase tissue transglutaminase
  • Type 3 transglutaminase epidermal transglutaminase
  • the source of the transglutaminase can be selected according to the particular use (e.g. site of implantation) of the medical implant material.
  • the medical implant material is to be used as artificial bone, it can be beneficial for the material to comprise a bone-derived transglutaminase.
  • any of transglutaminases can be used, and the origin and the production process thereof are not limited.
  • the transglutaminase can be derived from mammals or microorganisms. Moreover, humanized recombinant transglutaminase can also be used.
  • transglutaminases derived from mammals, such as guinea pig liver-derived transglutaminase, goat-derived transglutaminase and rabbit-derived transglutaminase, are available from Oriental Yeast Co., Ltd., Upstate USA Inc. and Biodesign International.
  • Other non-limiting examples of commercially available transglutaminase products include those produced by Ajinomoto Co. (Kawasaki, Japan), such as Activa TG-TI, Activa TG- FP, Activa TG-GS, Activa TG-RM, and Activa MP; and those produced by Yiming Biological Products Co. (Jiangsu, China), such as TG-B and TG-A.
  • the transglutaminase can be applied in any suitable form including, but not limited to, solutions, emulsions (oil-in-water, water-in-oil), aerosols, foams, ointments, pastes, lotions, powders, gels, hydrogels, hydrocolloids, creams, and any combinations thereof.
  • the transglutaminase is applied in the form of a powder.
  • the transglutaminase can optionally be in a pharmaceutical acceptable composition.
  • a pharmaceutically acceptable composition comprises a transglutaminase formulated together with one or more pharmaceutically acceptable carriers (additives) and/or diluents.
  • the pharmaceutically acceptable composition can also include any co-factors, such as salts, for the transglutaminase.
  • the bonding can be performed at any temperature.
  • the bonding is performed at an elevated temperature e.g., 30°C or higher, 35°C or higher, 40°C or higher, 45°C or higher, 50°C or higher, 55°C or higher, or 60°C or higher.
  • bonding is performed at room temperature, e.g., from about 15°C to about 25°C.
  • bonding is performed at a temperature of about 20°C to about 30°C.
  • an "effective amount of a transglutaminase” means the amount of transglutaminase which is effective to for adhering together two surfaces. Accordingly, in some embodiments, effective amount of a transglutaminase is from about ⁇ g to about lOOmg per cm 2 of contact surface area. In some embodiments, effective amount of a transglutaminase is selected from the group consisting of from about lmg to about 50mg per cm 2 , from about 5mg to about 40mg per cm 2 , from about lOmg to about 30mg per cm 2 , from about 15mg to about 25mg per cm 2 , or about 20mg per cm 2 .
  • the term “applying” refers to increasing the amount or activity of a transglutaminase at the desired site. Accordingly, the term “applying” embraces topical applications of transglutaminase at the desired site, increasing the expression of a host transglutaminase at the desired site, and/or increasing the activity of a transglutaminase at the desired site. Without wishing to be bound by a theory, one can increase the expression or activity of a host transglutaminase by applying a composition that increases the expression and/or activity of the transglutaminase. For example, Davies et al. (J. Biol. Chem.
  • transglutaminase activators include, but are not limited to, thrombin, TIG3 protein, calcium chloride, and sphingosylphosphorylcholine. Additionally, Sigma-Aldrich sells the Transglutaminase Assay Kit which can be used for screening activators of transglutaminase.
  • a glutamic acid rich polypeptide refers to polypeptides that have a plurality of glutamic acids. For example, at least 5% of the amino acids in the polypeptide are glutamic acid. In some embodiments, the glutamic acid rich polypeptide is casein.
  • an "effective amount of a glutamic acid rich polypeptide” means the amount of glutamic acid rich polypeptide which is effective to enhance bonding of two components with a transglutaminase. In some embodiments, effective amount of a glutamic acid rich polypeptide is from about ⁇ g to about lOOmg per cm 2 of contact surface area.
  • effective amount of a t glutamic acid rich polypeptide is selected from the group consisting of from about lmg to about 50mg per cm 2 , from about 5mg to about 40mg per cm 2 , from about lOmg to about 30mg per cm 2 , from about 15mg to about 25mg per cm 2 , or about 20mg per cm 2 .
  • Embodiments of the various aspects disclosed herein include a biomaterial.
  • biomaterial refers to any material that is biocompatible.
  • biocompatible material refers to any polymeric material that does not deteriorate appreciably and does not induce a significant immune response or deleterious tissue reaction, e.g., toxic reaction or significant irritation, over time when implanted into or placed adjacent to the biological tissue of a subject, or induce blood clotting or coagulation when it comes in contact with blood.
  • Suitable biocompatible materials include derivatives and copolymers of a polyimides, poly(ethylene glycol), polyvinyl alcohol, polyethyleneimine, and polyvinylamine, polyacrylates, polyamides, polyesters, polycarbonates, and polystyrenes.
  • the biomaterial comprises a carbohydrate-based polymer, wherein the polymer comprises at least one amino group. In some embodiments, the biomaterial comprises collagen or gelatin.
  • the biomaterial comprises a composite laminate material.
  • at least one layer of the composite laminate material comprises a protein and at least one layer of the composite laminate material comprises a carbohydrate.
  • Exemplary composite laminate materials amenable to the present invention are described for example in US Patent Application No. 13/819,391, content of which is incorporated herein by reference in its entirety.
  • the biomaterial is in form of a medical implant device.
  • a medical implant device refers to devices for implementation into a subject's body.
  • Exemplary medical implant devices include, but are not limited to, artificial tissues, artificial organs, prosthetic devices, drug delivery devices, wound dressings, fibers, nanoparticles, microparticles, foams, and sponges.
  • a medical implant device can be in any form including, but not limited to 3-D scaffolds, fibers, foams, sponges, films, and any combinations thereof.
  • a medical implant device can be used for permanent substitution of an organ (function).
  • the medical implant device is a foam or sponge.
  • the medical implant device comprises a nanoparticle or a microparticle.
  • the medical implant device is a wound dressing.
  • wound dressings include, but are not limited to bandages, gauzes, tapes, meshes, nets, adhesive plasters, films, membranes, and patches.
  • a wound dressing can comprise a composite material described herein.
  • the medical implant device is associated with a protein which is cross-linkable by a transglutaminase.
  • association with refers to a medical implant device which is coated with, includes, or comprises a transglutaminase linkable protein.
  • the medical implant device is coated with the transglutaminase linkable protein.
  • coated is meant that the transglutaminase linkable protein is applied to the surface of the medical implant device.
  • the medical implant device can be painted or sprayed with a solution comprising a transglutaminase linkable protein.
  • the medical implant device can be dipped in a solution of transglutaminase linkable protein solution.
  • the transglutaminase linkable protein can be covalently or non-covalently associated with the medical implant device, e.g. at the external surface of the medical implant device. Once associated with the medical implant device, the transglutaminase linkable protein provides means of attaching the medical implant device to a tissue or organ.
  • transglutaminase cross-linkable by a transglutaminase refers to a protein or polypeptide which serves as a substrate for a transglutaminase. Accordingly, a transglutaminase cross-linkable protein is or comprises a transglutaminase substrate.
  • transglutaminase substrate refers to a peptide or polypeptide sequence with an appropriate transglutaminase target for cross-linking.
  • a transglutaminase linkable protein is or comprises a transglutaminase substrate selected from the group consisting of aldolase A, glyceraldehyde-3 -phosphate dehydrogenase, phosphorylase kinase, crystalline, glutathione S-transferase, actin, myosin, troponin, ⁇ -tublin, tau, rho, histone, a-oxoglutarate dehydrogenase, ⁇ -lactoglobulin, cytochromes, erythrocyte band III, CD38, acetylcholine esterase, collagen, entactin, fibronectin, fibrin, silk, fibroin, fibrinogen, vitronectin, osteopontin, nidogen, laminin, LTBP-1, osteonectin, osteopontin, osteocalcin, thrombospondin, substance P, phospholipases A 2 , midkine, wheat ge
  • Peptide and polypeptide sequences with an appropriate transglutaminase target for cross-linking are known in the art. Non-limiting examples of such peptides are described, for example in U.S. Pat. No. 5,428,014; No. 5,939,385; and No. 7,208,171, content of all of which is incorporated herein by reference. U.S. Pat. No.
  • 5,428,014 describes biocompatible, bioadhesive, transglutaminase cross-linkable polypeptides wherein transglutaminase is known to catalyze an acyl-transfer reaction between the ⁇ -carboxamide group of protein-bound glutaminyl residues and the ⁇ -amino group of Lys residues, resulting in the formation of 8-(y-glutamyplysine isopeptide bonds.
  • U.S. Pat. No. 5,939,385 describes biocompatible, bioadhesive transglutaminase cross-linkable polypeptides.
  • U.S. Pat. No. 7,208,171 describes the rational design of transglutaminase substrate peptides.
  • the design strategy was based on maximizing the number of available acyl acceptor lysine-peptide substrates and acyl donor glutaminyl-peptide substrates available for transglutaminase cross-linking. Beyond this, the Lys and Glu substrate peptides were designed to possess basic features of known biomacromolecular and synthetic peptide substrates of transglutaminase.
  • the Glu substrate peptides contained 2-5 contiguous Glu residues, based on evidence that peptides become better transglutaminase substrates with increasing length of Glu repeats and that proteins containing two or more adjacent Glu residues are known to be good substrates.
  • a Leu residue was placed adjacent to the Glu near the C- terminus in several peptides, because this has been shown to result in a significant increase in Glu specificity.
  • Lys substrate peptides it has been shown that the composition and sequence of the amino acids adjacent to lysine residues in peptide and protein substrates can have an effect on the amine specificity.
  • a medical implant device can be fabricated from any biocompatible material.
  • biocompatible material refers to any polymeric material that does not deteriorate appreciably and does not induce a significant immune response or deleterious tissue reaction, e.g., toxic reaction or significant irritation, over time when implanted into or placed adjacent to the biological tissue of a subject, or induce blood clotting or coagulation when it comes in contact with blood.
  • Suitable biocompatible materials include derivatives and copolymers of a polyimides, poly(ethylene glycol), polyvinyl alcohol, polyethyleneimine, and polyvinylamine, polyacrylates, polyamides, polyesters, polycarbonates, and polystyrenes.
  • the medical implant device is fabricated from a material selected from the group consisting of carbohydrate polymers, proteins, silk fibroin, polydimethylsiloxane, polyimide, polyethylene terephthalate, polymethylmethacrylate, polyurethane, polyvinylchloride, polystyrene polysulfone, polycarbonate, polymethylpentene, polypropylene, a polyvinylidine fluoride, polysilicon, polytetrafluoroethylene, polysulfone, acrylonitrile butadiene styrene, polyacrylonitrile, polybutadiene, poly(butylene terephthalate), poly(ether sulfone), poly(ether ether ketones), poly(ethylene glycol), styrene-acrylonitrile resin, poly(trimethylene terephthalate), polyvinyl butyral, polyvinylidenedifluoride, poly(vinyl pyrrolidone),
  • a medical implant device can be fabricated from a biodegradable material, e.g., a biodegradable polymer.
  • biodegradable describes a material which can decompose under physiological conditions into breakdown products. Such physiological conditions include, for example, hydrolysis (decomposition via hydrolytic cleavage), enzymatic catalysis (enzymatic degradation), and mechanical interactions.
  • biodegradable also encompasses the term “bioresorbable”, which describes a substance that decomposes under physiological conditions to break down to products that undergo bioresorption into the host-organism, namely, become metabolites of the biochemical systems of the host organism.
  • biodegradable polymer refers to a polymer that at least a portion thereof decomposes under physiological conditions.
  • the polymer can thus be partially decomposed or fully decomposed under physiological conditions.
  • biodegradable polymers include, but are not limited to, polyanhydrides, polyhydroxybutyric acid, polyorthoesters, polysiloxanes, polycaprolactone, poly(lactic-co- glycolic acid), poly(lactic acid), poly(glycolic acid), and copolymers prepared from the monomers of these polymers.
  • the medical implant device is fabricated from a biocompatible, biodegradable material.
  • Suitable polymers which can be used for fabricating a medical implant device include, but are not limited to, one or a mixture of polymers selected from the group consisting of carbohydrate polymers; silk; glycosaminoglycan; fibrin; poly-ethyleneglycol (PEG); C2 to C4 polyalkylene glycols (e.g., propylene glycol); polyhydroxy ethyl methacrylate; polyvinyl alcohol; polyacrylamide; poly (N-vinyl pyrolidone); poly glycolic acid (PGA); poly lactic-co- glycolic acid (PLGA); poly e-carpolactone (PCL); polyethylene oxide; poly propylene fumarate (PPF); poly acrylic acid (PAA); hydrolysed polyacrylonitrile; polymethacrylic acid; polyethylene amine; polyanhydrides; polyhydroxybutyric acid; polyorthoesters; polysiloxanes; polycaprolactone; poly(lactic-co-glycolic acid); poly(lactic acid
  • the medical implant device is fabricated from a carbohydrate- based polymer.
  • the carbohydrate polymer is chitin, chitosan or a derivative thereof.
  • the medical implant device is fabricated from a transglutaminase linkable protein.
  • the medical implant device is fabricated from a composite material described herein.
  • Embodiments of the various aspects disclosed herein include a polymer.
  • the polymer comprises at least one group linkable by a transglutaminase.
  • An exemplary group that is linkable by a transglutaminase is an amino group.
  • the polymer comprises a protein, e.g., a transglutaminase linkable protein.
  • the polymer comprises a carbohydrate-based polymer.
  • Exemplary carbohydrate-based polymers include, but are not limited to, chitin and chitosan.
  • the polymer comprising at least one amino group includes a carbohydrate-based amino-polymer.
  • carbohydrate-based amino-polymer includes, but is not limited to, oligomers or polymers that contain monomers having the formula C m (H 2 0) n and at least one amino group, wherein m and n are > 3 and wherein m and n can be same or different. In some embodiments, m and n are independently 3, 4, 5, 6, or 7.
  • Carbohydrate-based amino-polymers include, but are not limited to, compounds such as oligosaccharides, polysaccharides, glycoproteins, glycolipids, and any combinations thereof.
  • Any carbohydrate-based polymer comprising an amino group that can crosslink with another reactive group of another molecule e.g., carboxamide group of a molecule or a polymer
  • a transglutaminase can be used in any embodiments of the methods and devices described herein.
  • the carbohydrate- based amino-polymer can comprise at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 or sugar monomers.
  • the carbohydrate-based amino-polymer can comprise sugar monomers independently selected from the group consisting of erythrose, threose, ribose, arabinose, xylose, lyxose, ribulose, xylulose, allose, altrose, glucose, mannose, gulose, idose, galactose, galactosamine, N-acetylgalactose, glucosamine, N-acetylglucosamine, sialic acid, talose, psicose, fructose, sorbose, tagatose, fucose, fuculose, rhamonse, sedoheptulose, octose, sulfoquinovose, glycosaminoglycan and nonose (neuraminic acid), wherein the sugar may be optionally substituted.
  • each sugar can independently have the L- or the D
  • the linkage between two sugar monomers can independently have a a- or ⁇ - configuration. Furthermore, the linkage between the two sugars can be 1— »3, 1— »4, 1— »5, or 1 ⁇ 6.
  • At least one (e.g., 1, 2, 3, or 4) hydroxyl group of the sugar monomer can be replaced by an amino group.
  • the hydroxyl group at position 2 of the sugar monomer can be replaced by an amino group.
  • the amino group can be optionally substituted with a C1-C6 alkyl or an acyl group.
  • C1-C6 alkyl groups include methyl, ethyl, propyl, butyl, and t-butyl.
  • the acyl group comprises acetyl.
  • the carbohydrate amino-polymer can comprise one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 or more) disaccharide, trisaccharide or tetrasaccharide monomers independently selected from the group consisting of sucrose, lactulose, lactose, maltose, trehalose, cellobiose, kojibiose, nigerose, isomaltose, ⁇ , ⁇ -Trehalose, ⁇ , ⁇ -Trehalose, sophorose, laminaribiose, gentibiose, turanose, maltulose, palatinose, gentibiulose, mannobiose, melibiose, rutinose, rutinulose, xylobiose, raffinose, melezitose, acarbose and stachyose.
  • disaccharide e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 or more
  • oligosaccharide refers without limitation to several (e.g., five to ten) covalently linked monosaccharide units.
  • polysaccharide refers without limitation to many (e.g., eleven or more) covalently linked sugar units. Polysaccharides can have molecular masses up to millions of Daltons.
  • Exemplary oligosaccharides and polysaccharides include, but are not limited to, fructooligosaccharide, galactooligosaccharides, mannanoligosaccharides, glycogen, starch (amylase, amylopectin), glycosaminoglycans (e.g., hyaluronic acid, chondroitin-4-sulfate, chondroitin-6-sulfate, dermatan sulfate, keratin sulfate, and heparin), cellulose, beta-glucan (e.g., zymosan, lentinan, sizofiran), maltodextrin, inulin, levan beta (2— »6), chitin, and chitosan.
  • fructooligosaccharide e.g., galactooligosaccharides, mannanoligosaccharides, glycogen, starch (amylase, amylopectin), glycos
  • the carbohydrate- based amino-polymer can comprise chitin or a derivative thereof.
  • chitin derivative comprises chitosan (a-(l-4) 2-amino-2-deoxy-P-D-glucan) or a derivative thereof.
  • chitosan can also include all derivatives of chitin, or poly-N-acetyl-D-glucosamine (including all polyglucosamine and oligomers of glucosamine materials of different molecular weights), in which at least about 20% of the acetyl groups (e.g., at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or more, including 100% of the acetyl groups) have been removed through hydrolysis.
  • the acetyl groups e.g., at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or more, including 100% of the acetyl groups
  • chitosans are a family of cationic, binary heteropoly saccharides composed of (1— »4)-linked 2-acetamido-2-deoxy-(3-D-glucose (GlcNAc, A-unit) and 2-amino-2-deoxyP-D-glucose, (GlcN; D-unit) (Varum K. M. et al., Carbohydr. Res., 1991, 217: 19-27; Sannan T. et al., Macromol. Chem., 1776, 177: 3589-3600).
  • Chitosan, chitosan derivatives or salts (e.g., but not limited to, nitrate, phosphate, sulphate, hydrochloride, glutamate, lactate or acetate salts) of chitosan can be used in any embodiments of the methods and devices described herein.
  • Any art-recognized chitosan and derivatives thereof, e.g., the ones described in the U.S. Patent No. 7,125,967 and 7,288,532, the content of which are incorporated herein by reference, can be provided as the polymer comprising an amino group used in some embodiments of the methods and devices described herein.
  • chitosan derivatives are intended to include ester, ether or other derivatives formed by bonding of acyl and/or alkyl groups with OH groups, but not the H 2 groups, of chitosan. Examples are O-alkyl ethers of chitosan and O-acyl esters of chitosan.
  • chitosan include, but are not limited to, N-(aminoalkyl) chitosans, succinyl chitosans, quteraminated chitosans, N-acylated chitosans (e.g., caproyl chitosan, octanoyl chitosan, myristoyl chitosan, and palmitoyl chitosan), N-methylene phosphonic chitosans, N-lauryl -N-methylene phosphonic chitosans, N-lauryl-carboxymethyl chitosans, N-alkyl-O-sulfated chitosans, thiolated chitosans (e.g., chitosan-2-iminthiolane, chitosan-4-thiobutylamidine, and chitosan-thioglycolic acid), trimethylchitosan, and phosphorylated chitosans,
  • Modified chitosans e.g., those chitosans conjugated to polyethylene glycol, can also be utilized in any embodiments of the methods and devices described herein.
  • Chitosans of various viscosity can be obtained from various sources, e.g., chitosan produced by deacetylation of chitin, which, for example, can be obtained from the structural element in the exoskeleton of crustaceans (such as crabs and shrimp) and cell walls of fungi, or from commercial sources including, but not limited to, PRONOVA Bioamino-polymer, Ltd. (UK); SEIGAGAKU America Inc. (Maryland, USA); MERON (India) Pvt, Ltd. (India); VANSON Ltd. (Virginia, USA); and AMS Biotechnology Ltd. (UK).
  • sources e.g., chitosan produced by deacetylation of chitin, which, for example, can be obtained from the structural element in the exoskeleton of crustaceans (such as crabs and shrimp) and cell walls of fungi, or from commercial sources including, but not limited to, PRONOVA Bioamino-polymer, Ltd. (UK); SEIGA
  • Any carbohydrate-based polymer comprising at least one amino group can be used in any embodiments of the compositions and methods described herein.
  • One of skill in the art is well aware of synthetic methods which can be used for the synthesis of carbohydrate-based amino-polymers. See for example, Stick, R.V., Carbohydrates: The Sweet Molecules of Life.; Academic Press, pp 113-177 (2002); Crich, D. & Dudkin V., J. Am. Chem. Soc, 123 :6819-6825 (2001); Garegg, P. J., Chemtracts-Org. Chem., 5:389 (1992); Mayer, T. G, Kratzer, B. & Schmidt, R.
  • Embodiments of the various aspects disclosed herein include a protein.
  • the protein can be selected from the group consisting of collagen, gelatin, perculin, abductin, fibrin, fibroin, elastin, resilin, fibronectin, fibrinogen, keratin, titin, actin, Arp2/3, coronin, dystrophin, FtsZ, myosin, spectrin, Tau (protein), tubulin, F-spondin, Pikachurin, protein fragments, synthetic peptides, genetically expressed portions of proteins, fragments thereof, and any combinations thereof.
  • the protein is collagen or gelatin.
  • the invention provides a three component composition.
  • the three components are in flowable form.
  • the first component is a polymer comprising an amino group, i.e., an amino polymer;
  • the second component is a transglutaminase; and
  • the third component is a glutamic acid rich polypeptide.
  • the three components can be in separate formulations, in one formulation or two in one formulation and the other in a separate formulation.
  • the amino polymer is in a separate formulation than the transglutaminase and the glutamic acid rich polypeptide.
  • the transglutaminase and the glutamic acid rich polypeptide are in one formulation.
  • a method for bonding or adhering a biomaterial to a target surface the method
  • transglutaminase and a glutamic acid rich polypeptide comprising applying an effective amount of a transglutaminase and a glutamic acid rich polypeptide to a target surface and contacting a biomaterial to the target surface where the transglutaminase and the glutamic acid rich polypeptide have been applied.
  • biomaterial comprises a protein or a carbohydrate-based polymer comprising at least one amino group.
  • biomaterial comprises a composite laminate material comprising a layer of carbohydrate-based polymer and a layer of protein.
  • the protein of the composite laminate material comprises collagen, gelatin or silk fibroin.
  • the medical implant device is selected from the group consisting of artificial tissues, artificial organs, prosthetic devices, drug delivery devices, wound dressings, films, foams, sponges, scaffolds, meshes, hemostatic materials, and any combinations thereof.
  • the medical implant device is a wound dressing selected from the group consisting of bandages, gauzes, tapes, meshes, nets, adhesive plasters, films, membranes, patches, microparticles, nanoparticles, and any combinations thereof.
  • the target surface is a surface of a tissue or organ.
  • the target surface is a surface of a hepatic, cardiac, intestinal, pulmonary or dermal tissue.
  • the wound is selected from the group consisting of cuts and lacerations, surgical incisions, punctures, grazes, scratches, compression wounds, abrasions, friction wounds, chronic wounds, ulcers, thermal effect wounds, chemical wounds, wounds resulting from pathogenic infections, skin graft/transplant donor and recipient sites, immune response conditions, oral wounds, stomach or intestinal wounds, damaged cartilage or bone, amputation sites, corneal lesions and lung punctures.
  • the transglutaminase is a mammalian or microbial transglutaminase.
  • a method for promoting wound healing comprising applying a
  • transglutaminase and a glutamic acid rich polypeptide to a surface of a wound.
  • the polymer is in form of a solution, an emulsion, an aerosol, a foam, an ointment, a paste, a lotion, a powder, a gel, a hydrogel, a bandage, a gauze, a tape, a mesh, a net, an adhesive plaster, a film, a membrane, a patch, a microparticle, a nanoparticle or any combinations thereof.
  • the wound is selected from the group consisting of cuts and lacerations, surgical incisions, punctures, grazes, scratches, compression wounds, abrasions, friction wounds, chronic wounds, ulcers, thermal effect wounds, chemical wounds, wounds resulting from pathogenic infections, skin graft/transplant donor and recipient sites, immune response conditions, oral wounds, stomach or intestinal wounds, damaged cartilage or bone, amputation sites, corneal lesions and lung punctures.
  • transglutaminase is a mammalian or microbial transglutaminase.
  • a method for forming a coating layer on a target surface comprising:
  • polymer is a protein or a carbohydrate- based polymer comprising at least one amino group.
  • carbohydrate-based polymer comprises chitin or chitosan.
  • the target surface is a surface of a hepatic, cardiac, intestinal, pulmonary or dermal tissue.
  • the wound is selected from the group consisting of cuts and lacerations, surgical incisions, punctures, grazes, scratches, compression wounds, abrasions, friction wounds, chronic wounds, ulcers, thermal effect wounds, chemical wounds, wounds resulting from pathogenic infections, skin graft/transplant donor and recipient sites, immune response conditions, oral wounds, stomach or intestinal wounds, damaged cartilage or bone, amputation sites, corneal lesions and lung punctures.
  • the transglutaminase is a mammalian or microbial transglutaminase.
  • a three component composition comprising as a first component a polymer in a flowable form, wherein the polymer comprises an amino group; as a separate second component a transglutaminase in a flowable form; and as a separate third component a glutamic acid rich polypeptide in a flowable form.
  • composition of paragraph 51 wherein at least two of the first, second and the third component are formulated together.
  • composition of any of paragraphs 51-53, wherein the flowable form includes a liquid, powder, or a combination thereof.
  • the composition of any of paragraphs 51-56, wherein the second and the third component are provided in a powder form.
  • the composition of any of paragraphs 51-56, wherein the second and the third component are provided in a liquid form.
  • the composition of paragraph 57 or 58, wherein the first component is in a liquid form.
  • composition of paragraph 57 or 58 wherein the first component is in form of a liquid.
  • the composition of paragraph 61 wherein the first component is in form of an aerosol.
  • the composition of any of paragraphs 51-64 wherein the glutamic acid rich polypeptide is casein. 66.
  • composition of paragraph 66, wherein the carbohydrate-based polymer comprises chitin or chitosan.
  • composition of paragraph 66, wherein the protein is collagen or gelatin.
  • glutamic acid rich polypeptide are applied concurrently to the target surface.
  • compositions, methods, and respective component(s) thereof that are essential to the invention, yet open to the inclusion of unspecified elements, whether essential or not.
  • the terms “comprising” and “comprises” include the terms “consisting of and “consisting essentially of.”
  • the term "consisting essentially of” refers to those elements required for a given embodiment. The term permits the presence of additional elements that do not materially affect the basic and novel or functional characteristic(s) of that embodiment of the invention.
  • compositions, methods, and respective components thereof as described herein, which are exclusive of any element not recited in that description of the embodiment.
  • “decrease”, “reduced”, “reduction”, “decrease” or “inhibit” are all used herein generally to mean a decrease by a statistically significant amount.
  • “reduced”, “reduction” or “decrease” or “inhibit” means a decrease by at least 10% as compared to a reference level, for example a decrease by at least about 20%, or at least about 30%), or at least about 40%, or at least about 50%, or at least about 60%, or at least about 70%, or at least about 80%, or at least about 90% or up to and including a 100% decrease (e.g. absent level as compared to a reference sample), or any decrease between 10-100%) as compared to a reference level.
  • the terms “increased”, “increase” or “enhance” or “activate” are all used herein to generally mean an increase by a statically significant amount; for the avoidance of any doubt, the terms “increased”, “increase” or “enhance” or “activate” means an increase of at least 10% as compared to a reference level, for example an increase of at least about 20%, or at least about 30%), or at least about 40%, or at least about 50%, or at least about 60%, or at least about 70%, or at least about 80%, or at least about 90% or up to and including a 100% increase or any increase between 10-100%) as compared to a reference level, or at least about a 2-fold, or at least about a 3-fold, or at least about a 4-fold, or at least about a 5-fold or at least about a 10-fold increase, or any increase between 2-fold and 10-fold or greater as compared to a reference level.
  • the term "statistically significant” or “significantly” refers to statistical significance and generally means a two standard deviation (2SD) below normal, or lower, concentration of the marker.
  • the term refers to statistical evidence that there is a difference. It is defined as the probability of making a decision to reject the null hypothesis when the null hypothesis is actually true. The decision is often made using the p-value.
  • the terms "effective” and “effectiveness” includes both pharmacological effectiveness and physiological safety.
  • Pharmacological effectiveness refers to the ability of the treatment to result in a desired biological effect in the patient.
  • Physiological safety refers to the level of toxicity, or other adverse physiological effects at the cellular, organ and/or organism level (often referred to as side-effects) resulting from administration of the treatment.
  • Less effective means that the treatment results in a therapeutically significant lower level of pharmacological effectiveness and/or a therapeutically greater level of adverse physiological effects.
  • a "subject” means a human or animal. Usually the animal is a vertebrate such as a primate, rodent, domestic animal or game animal. Primates include chimpanzees, cynomologous monkeys, spider monkeys, and macaques, e.g., Rhesus. Rodents include mice, rats, woodchucks, ferrets, rabbits and hamsters.
  • Domestic and game animals include cows, horses, pigs, deer, bison, buffalo, feline species, e.g., domestic cat, canine species, e.g., dog, fox, wolf, avian species, e.g., chicken, emu, ostrich, and fish, e.g., trout, catfish and salmon.
  • Patient or subject includes any subset of the foregoing, e.g., all of the above, but excluding one or more groups or species such as humans, primates or rodents.
  • the subject is a mammal, e.g., a primate, e.g., a human.
  • the terms, "patient” and “subject” are used interchangeably herein.
  • the terms, "patient” and “subject” are used interchangeably herein.
  • a subject can be male or female.
  • the subject is a mammal.
  • the mammal can be a human, non -human primate, mouse, rat, dog, cat, horse, or cow, but are not limited to these examples. Mammals other than humans can be advantageously used as subjects that represent animal models of disorders associated with autoimmune disease or inflammation.
  • the methods and compositions described herein can be used to treat domesticated animals and/or pets.
  • contact surface area is meant the surface area of the target surface needed for bonding.
  • the contact surface area can mean the size of the wound and/or the total exposed area of the wound.
  • the term "pharmaceutically acceptable” refers to those compounds, materials, compositions, and/or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit/risk ratio.
  • the term "pharmaceutically-acceptable carrier” means a pharmaceutically-acceptable material, composition or vehicle, such as a liquid or solid filler, diluent, excipient, manufacturing aid (e.g., lubricant, talc magnesium, calcium or zinc stearate, or steric acid), or solvent encapsulating material, involved in carrying or transporting the subject compound from one organ, or portion of the body, to another organ, or portion of the body.
  • manufacturing aid e.g., lubricant, talc magnesium, calcium or zinc stearate, or steric acid
  • solvent encapsulating material involved in carrying or transporting the subject compound from one organ, or portion of the body, to another organ, or portion of the body.
  • Each carrier must be “acceptable” in the sense of being compatible with the other ingredients of the formulation and not injurious to the patient.
  • materials which can serve as pharmaceutically-acceptable carriers include: (1) sugars, such as lactose, glucose and sucrose; (2) starches, such as corn starch and potato starch; (3) cellulose, and its derivatives, such as sodium carboxymethyl cellulose, methylcellulose, ethyl cellulose, microcrystalline cellulose and cellulose acetate; (4) powdered tragacanth; (5) malt; (6) gelatin; (7) lubricating agents, such as magnesium stearate, sodium lauryl sulfate and talc; (8) excipients, such as cocoa butter and suppository waxes; (9) oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; (10) glycols, such as propylene glycol; (11) polyols, such as glycerin, sorbitol, mannitol and polyethylene glycol (PEG); (12) esters, such as ethyl
  • wetting agents, coloring agents, release agents, coating agents, sweetening agents, flavoring agents, perfuming agents, preservative and antioxidants can also be present in the formulation.
  • excipient e.g., pharmaceutically acceptable carrier or the like are used interchangeably herein.
  • Example 1 Direct bonding of chitosan biomaterials to tissue using transglutaminase for surgical repair or device implantation.
  • Natural biomaterials such as chitosan and collagen, are useful for biomedical applications because they are biocompatible, mechanically robust and biodegradable, but it is difficult to rapidly and tightly bond them to living tissues.
  • microbial enzyme transglutaminase mTG was used to rapidly ( ⁇ 5 min) bond chitosan and collagen biomaterials to the surfaces of hepatic, cardiac and dermal tissues, as well as to functionalized polydimethylsiloxane (PDMS) materials that are used in medical products.
  • mTG-bondedshrilk patches composed of a chitosan-fibroin laminate effectively sealed intestinal perforations, and a newly developed two-component mTG- bonded chitosan spray effectively repaired ruptures in a breathing lung when tested ex vivo.
  • the mechanical strength of mTG- catalyzed chitosan adhesive bonds were comparable to those generated by commonly used surgical glues.
  • TG transglutaminase
  • Tissue-derived TG has been employed to bond pieces of cartilage [see for example, Jiirgensen, K., et al., A New Biological Glue for Cartilage-Cartilage Interfaces: Tissue Transglutaminase 79, 85-193 (1997)] and microbial TG (mTG) has been used to crosslink protein gels, including fibrin and gelatin gels, which were shown to exhibit improved cell attachment and resistance to protease degradation. See for example, Chau, D.Y. S., et al., The cellular response to transglutaminase-cross-linked collagen. Biomaterials, 26(33), 6518-6529 (2005).
  • TGs also have been investigated for the crosslinking of proteins to non-proteinaceous molecules that require functional groups, such as hyaluronic acid [see for example, Picard, J., S. Giraudier, and Larreta-Garde, V. Controlled remodeling of a protein-poly saccharide mixed gel: examples of gelatin-hyaluronic acid mixtures. Soft Matter, 5(21), 4198-4205 (2009)] and peptide-modified polyethyleneglycol (PEG). See for example, Sperinde, J.J. and Griffith, L. G. Control and Prediction of Gelation Kinetics in Enzymatically Cross-Linked Poly(ethylene glycol) Hydrogels. Macromolecules, 33(15), 5476-5480 (2000).
  • functional groups such as hyaluronic acid
  • PEG peptide-modified polyethyleneglycol
  • mTG was chosen rather than tissue-derived TG in the discussed experiment because it can be produced much more efficiently at lower cost, and it does not require the presence of calcium ions to be activated, which enables many applications not supported by tissue- derived TG.
  • the results show that mTG can be used to bond chitosan and collagen materials to organic substrates including different tissue types as well as inorganic substrates modified to contain sterically available amine groups. However, this process is only generalizable when the mTG preparation contains casein which is rich in glutamic acid residues.
  • carbohydrates e.g., maltodextrin, saccharose, or mannose
  • carbohydrates e.g., maltodextrin, saccharose, or mannose
  • proteins are added to protect mTG against degradation by extracellular proteolytic enzymes. See for example, Junqua, M., et al., Optimization of microbial transglutaminase production using experimental designs. Applied Microbiology and Biotechnology, 48(6), 730-734 (1997).
  • mTG as a bonding reagent for chitosan materials
  • one preparation contained only maltodextrin (mTG+Ma) and another contained both this carbohydrate and casein (mTG+Ca).
  • the mTG preparation was applied as a powder to one of the surfaces before the two materials were brought together without applying pressure. Since both surfaces are hydrated and hydrophilic, capillary forces held the films in close contact. 10 minutes after mTG application, the adhesion force required to separate two mTG-bonded films was quantified using a standard t-peel test protocol (ASTM D1876).
  • Implanted medical devices, electrochemical sensors, and actuators require strong and preferentially seamless interfaces between inorganic materials and living tissues, as do recently developed microfluidic Organ-on-a-chip' culture devices. See for example, Huh, D., et al, Reconstituting Organ-Level Lung Functions on a Chip. Science, 328(5986), 1662-1668 (2010); Bhatia, S.N. and Ingber, D. E. Microfluidic organs-on-chips. Nat. Biotech, 32(8), 760-772 (2014).
  • Polydimethylsiloxane is a silicone polymer broadly used in medical devices, as well as microfluidic organ-on-a-chip devices, which does not normally serve as a substrate for mTG-mediated adhesion because it lacks endogenous reactive amine groups.
  • mTG catalyzed adhesion of engineered organic biofilms to include bonding of these biofilms to PDMS was explored by first functionalizing its surface with amine groups using 3- Triethoxysilylpropylamine (APTES) before addition of mTG (FIG. 2a). Similar bonding studies were performed using collagen or chitosan films with mTG+Ma versus mTG+Ca, and adhesion strength was measured.
  • APTES 3- Triethoxysilylpropylamine
  • the three materials were assayed with the "Standard Test Method for Burst Strength of Surgical Sealants" (ASTM F2392), a standard industrial protocol that measures the ultimate pressure (pressure necessary to break the sealing) of a surgical patch covering a 3 mm circular perforation on standard tissue mimic surface (e.g., collagen film #320; Nippi, Inc., Japan) (FIG. 3a).
  • ASTM F2392 Standard Test Method for Burst Strength of Surgical Sealants
  • maximum pressure reached among all the samples is also reported, as it is indicative of bonding strength produced by each adhesion method independently of the user's skills.
  • mTG+Ca method was used to attach chitosan films to whole porcine liver, heart and skin explants. Chitosan films were covered with the mTG powder and placed on the surface of the tissue for 10 min at room temperature. Again no pressure was applied during the reaction time, and instead the film conformed to the shape and held itself in place because of the small amounts of water it holds on its surface and the action of resultant capillary forces.
  • chitosan foams can be similarly bonded to tissues by using mTG+Ca to adhere chitosan foams to an irregularly shaped 1 cm defect in an explanted latissimus dorsi muscle from a domestic pig. Again, these foams exhibited firm attachment to the tissue boundaries within 10 minutes after application, and the adhesion between the foam and tissue was virtually seamless with the border being difficult to detect when analyzed with photomicroscopy (FIG. 3e).
  • mTG+Ca was used to bond a 3 x 3 cm 2 chitosan-fibroin laminate film, known as Shrilk [see for example, Fernandez, J.G. and Ingber, D. E., Unexpected Strength and Toughness in Chitosan-Fibroin Laminates Inspired by Insect Cuticle. Advanced Materials, 24(4), 480-484 (2012)], over the hole by placing the mTG powder-coated film in contact with the inner surface of the intestinal wall surrounding the damaged area (e.g., mimicking placement via endoscopic surgery). A burst test was performed 10 minutes later by clamping both ends of the intestinal segment onto the end of a hollow tube and flowing saline into the lumen to increase intraluminal hydraulic pressure.
  • Bonding of porous chitosan foams to dermis might be useful for mechanical treatment strategies used for large non-healing wounds, such as Vacuum Assisted Closure (VAC) therapy, which would benefit from use of bioabsorbable porous scaffolds that provide both good physical properties and strong attachment to the wound site.
  • VAC Vacuum Assisted Closure
  • bioabsorbable porous scaffolds that provide both good physical properties and strong attachment to the wound site.
  • the discussed experiment demonstrates how mTG preparations containing casein can be used to bond chitosan to materials composed of proteins, such as collagen and living tissues, as well as to inorganic surfaces of polymers that are used in medical and microfluidic devices, such as PDMS.
  • the rapid action and the versatility of the bonding process make it suitable for a broad number of applications, ranging from surgical sealants to coatings for implantable medical devices or sensor/actuators to better integrate biomaterials and living tissues within microfluidic devices or BioMEMS.
  • the ability of mTG to rapidly and strongly bond chitosan and other biomaterials to both living tissues and inorganic surfaces offers a new way to seamlessly integrate living and non-living materials.
  • microbial transglutaminase (mTG) preparations containing Streptoverticillium calcium-independent TG were obtained from Ajinomoto Food Ingredients LLC (Chicago, USA).
  • One preparation contained approximately 1% (w/w) enzyme stabilized in maltodextrin, whereas the other was stabilized using maltodextrin (0.39 w/w) and casein (0.6 w/w).
  • Enzymatic preparations were stored in a powder form to facilitate transport of mTG at room and higher temperatures, and because this formulation could enable its use across a broad spectrum of applications in the future (e.g., first aid and battlefield assistance).
  • mTG preparations were used as a 3% (w/v) solution in 4% (w/v) NaOH to maintain their activity. See for example, Yokoyama, K., N. Nio, and Kikuchi, Y. Properties and applications of microbial transglutaminase. Applied Microbiology and Biotechnology, 64(4), 447-454 (2004). Although higher mTG concentrations enhanced the film- forming capabilities of the spray components, they were not useful for spraying due to micelle formation in the solution.
  • Chitosan films and foams were produced as reported before. See for example, Fernandez, J.G. and Ingber, D.E. Bioinspired Chitinous Material Solutions for Environmental Sustainability and Medicine. Advanced Functional Materials, 23(36), 4454-4466 (2013); Fernandez, J.G. and Ingber, D. E., Unexpected Strength and Toughness in Chitosan-Fibroin Laminates Inspired by Insect Cuticle. Advanced Materials, 24(4), 480-484 (2012). Chitosan films were used to investigate bonding strength using standardized industrial methods and ASTM protocols; chitosan foams were used to investigate tissue defect filling.
  • Chitosan films were produced by solvent evaporation casting of a 2% (w/v) solution of 80% deacetylated chitosan in a 1% (v/v) acetic acid solution. Films were neutralized with a 4% NaOH solution (w/v).
  • the method for producing Shrilk films composed of a laminate of layers of chitosan and fibroin has been previously described in detail. See for example, Fernandez, J.G. and Ingber, D. E., Unexpected Strength and Toughness in Chitosan-Fibroin Laminates Inspired by Insect Cuticle. Advanced Materials, 24(4), 480-484 (2012).
  • Chitosan foams were made by freeze drying a 1% (w/v) solution of chitosan in a 0.5% (v/v) acetic acid solution, neutralizing it in 4% (w/v) NaOH, and intensely washing in double ionized water. Due to the randomness of the foam structure, this configuration is not suitable for a standardized measurement of bonding strength, and studies with living tissues were carried out instead, as described in the results.
  • Collagen films were produced from the Collagen casing #320 (Nippi, Inc., Japan) standard for ASTM protocols (e.g. ASTM F2392). Chitosan films strengthened with fibroin (i.e.shrilk) were produced by sequential deposition and neutralization of the components, as described previously. See for example, Fernandez, J.G. and Ingber, D. E., Unexpected Strength and Toughness in Chitosan-Fibroin Laminates Inspired by Insect Cuticle. Advanced Materials, 24(4), 480-484 (2012).
  • Adhesion measurements were performed using two ASTM methods.
  • ASTM D1876 (t-peel test) experiments were performed with an Instron 3342 instrument (500N, Instron, USA) to measure the strength of adhesion of films bonded to flat surfaces.
  • Studies were carried out on biopolymer films, collagen substrates, and tissue s(i.e. skin, lung, heart, and muscle) shaped in rectangular (1 cm wide x 6 cm long) strips. Both surfaces of the adhesion test were fixed to an aluminum support (200 ⁇ thick) to avoid the effect of the film and substrate stretching when measuring surface adhesion; pulling speed was 10 mm/min.
  • the treated side of the films was placed in direct contact, but they were not subjected to pressure during the reaction time, and instead were held in place by capillary forces.
  • Lung tests with the double spray were performed using explanted lungs of domestic pig (sus domesticus) obtained from a local slaughterhouse. A deep hole (3 x 1.5 cm) was formed on the surface of one lung, and the trachea was connected to an external air pump. Continuous cyclic airflow to fully inflate and deflate the lungs was provided to mimic breathing motions of the lung. Tests were carried out twice on the same pair of lungs. Intestinal tests were performed by attaching both ends of a 15 cm section of explanted small intestine from a domestic pig to 3 cm diameter tube.
  • a 1 cm hole was incised in the intestine wall and subsequently repaired using mTG in powder form to attach a 3 cm patch of dvslk (chitosan-fibroin laminate film; see for example, Fernandez, J.G. and Ingber, D. E., Unexpected Strength and Toughness in Chitosan- Fibroin Laminates inspired by Insect Cuticle. Advanced Materials, 24(4), 480-484 (2012)) from inside the lumen of the intestine.
  • the system was filled with pressurized water containing blue dye (to enhance contrast) and pressure was measured using a differential pressure sensor (Pasco, California, USA).

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Abstract

L'invention concerne des compositions et des procédés pour lier des biomatériaux à des tissus et à des surfaces inorganiques.
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Cited By (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
RU2665155C1 (ru) * 2018-01-10 2018-08-28 федеральное государственное бюджетное образовательное учреждение высшего образования "Приволжский исследовательский медицинский университет" Министерства здравоохранения Российской Федерации Способ доставки биологически активных веществ в скаффолд
WO2021247774A1 (fr) * 2020-06-02 2021-12-09 The Johns Hopkins University Gel destiné à être utilisé en endoscopie gastro-intestinale et autres utilisations endodermiques, épidermiques et muqueuses
US11344397B2 (en) 2015-06-30 2022-05-31 Tela Bio, Inc. Corner-lock stitch patterns
US11369464B2 (en) 2015-07-21 2022-06-28 Tela Bio, Inc Compliance control stitching in substrate materials
US11446130B2 (en) 2019-03-08 2022-09-20 Tela Bio, Inc. Textured medical textiles
US11464616B2 (en) 2016-04-26 2022-10-11 Tela Bio, Inc. Hernia repair grafts having anti-adhesion barriers
US11590262B2 (en) 2018-03-09 2023-02-28 Tela Bio, Inc. Surgical repair graft
US11771769B2 (en) 2017-11-10 2023-10-03 Cocoon Biotech Inc. Ocular applications of silk-based products

Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US1893608A (en) * 1929-08-03 1933-01-10 Atwood Francis Clarke Casein solution and method of making the same
US3421918A (en) * 1963-11-27 1969-01-14 Borden Inc Alcohol soluble casein product for setting human hair
US5736132A (en) * 1993-06-03 1998-04-07 Orthogene, Inc. Method of promoting adhesion between tissue surfaces
US20020106423A1 (en) * 2001-01-25 2002-08-08 Nutricepts, Inc. Shaped cheese reconstruction with transglutaminase
US20040030408A1 (en) * 2000-05-12 2004-02-12 Martin Griffin Medical implant materials
US20130287836A1 (en) * 2010-08-30 2013-10-31 President And Fellows Of Harvard College High strength chitin composite material and method of making
WO2014014370A2 (fr) * 2012-07-20 2014-01-23 Politechnika Gdanska Procédé d'obtention d'une solution aqueuse de chitosane, composition de chitosane, aérosol de chitosane, procédé de production d'une membrane d'hydrogel à base de chitosane et procédé de production d'un matériau biopolymère de type chitosane-protéine
US20150359857A1 (en) * 2014-04-07 2015-12-17 Biomedica Management Corporation Aerosolized fibrin hemostat

Patent Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US1893608A (en) * 1929-08-03 1933-01-10 Atwood Francis Clarke Casein solution and method of making the same
US3421918A (en) * 1963-11-27 1969-01-14 Borden Inc Alcohol soluble casein product for setting human hair
US5736132A (en) * 1993-06-03 1998-04-07 Orthogene, Inc. Method of promoting adhesion between tissue surfaces
US20040030408A1 (en) * 2000-05-12 2004-02-12 Martin Griffin Medical implant materials
US20020106423A1 (en) * 2001-01-25 2002-08-08 Nutricepts, Inc. Shaped cheese reconstruction with transglutaminase
US20130287836A1 (en) * 2010-08-30 2013-10-31 President And Fellows Of Harvard College High strength chitin composite material and method of making
WO2014014370A2 (fr) * 2012-07-20 2014-01-23 Politechnika Gdanska Procédé d'obtention d'une solution aqueuse de chitosane, composition de chitosane, aérosol de chitosane, procédé de production d'une membrane d'hydrogel à base de chitosane et procédé de production d'un matériau biopolymère de type chitosane-protéine
US20150359857A1 (en) * 2014-04-07 2015-12-17 Biomedica Management Corporation Aerosolized fibrin hemostat

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
"Material Safety Data Sheet, uasein, tech.", ACROS ORGANICS, 16 July 1996 (1996-07-16) *
"Safety Data Sheet (Chitosan", PARCHEM, 24 June 2015 (2015-06-24), pages 1 *

Cited By (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US11344397B2 (en) 2015-06-30 2022-05-31 Tela Bio, Inc. Corner-lock stitch patterns
US11864987B2 (en) 2015-06-30 2024-01-09 Tela Bio, Inc. Corner-lock stitch patterns
US11369464B2 (en) 2015-07-21 2022-06-28 Tela Bio, Inc Compliance control stitching in substrate materials
US11464616B2 (en) 2016-04-26 2022-10-11 Tela Bio, Inc. Hernia repair grafts having anti-adhesion barriers
US11771769B2 (en) 2017-11-10 2023-10-03 Cocoon Biotech Inc. Ocular applications of silk-based products
RU2665155C1 (ru) * 2018-01-10 2018-08-28 федеральное государственное бюджетное образовательное учреждение высшего образования "Приволжский исследовательский медицинский университет" Министерства здравоохранения Российской Федерации Способ доставки биологически активных веществ в скаффолд
US11590262B2 (en) 2018-03-09 2023-02-28 Tela Bio, Inc. Surgical repair graft
US11446130B2 (en) 2019-03-08 2022-09-20 Tela Bio, Inc. Textured medical textiles
WO2021247774A1 (fr) * 2020-06-02 2021-12-09 The Johns Hopkins University Gel destiné à être utilisé en endoscopie gastro-intestinale et autres utilisations endodermiques, épidermiques et muqueuses

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