WO2011123665A1

WO2011123665A1 - Methods and compositions for the treatment of burns and wounds

Info

Publication number: WO2011123665A1
Application number: PCT/US2011/030759
Authority: WO
Inventors: Michael Rice; Aby J. Mathew; Dominic M. Clarke; Ian B. Nicoud
Original assignee: Biolife Solutions, Inc.
Priority date: 2010-03-31
Filing date: 2011-03-31
Publication date: 2011-10-06

Abstract

Methods are provided for treating wounds, including but not limited to wounds caused by trauma, medical intervention, and/or burns. The methods can include contacting the wound with a treatment solution, such as HYPOTHERMOSOL® and equivalents thereof, and maintaining the treatment solution in the area of the wound for a period of time. In some embodiments, the treatment solution is chilled below room temperature prior to administration.

Description

METHODS AND COMPOSITIONS FOR THE

TREATMENT OF BURNS AND WOUNDS

REFERENCE TO RELATED APPLICATIONS

This application claims priority to and the benefit of U.S. Provisional

Application No. 61/319,319, filed on March 31, 2010, the entire contents of which are incorporated by reference herein.

BACKGROUND

Approximately 3.5 million thermal injuries occur in the United States every year, with over 20% requiring hospitalization and specialized care. Traber et al., "Animal Models of Burn Injury," Surgical Research, Ch 31, Academic Press, San Diego, CA (2001). In addition, millions of traumatic injuries (e.g., accidents) and medically-related injuries (e.g., surgery) occur each year. The primary goals of burn and wound care are analgesia, antisepsis, prevention of wound progression, expedited closure, restoration of function, and reduction of scarring.

Current treatments for traumatic injuries, burns and other wounds are limited and often have adverse consequences, such as scarring. Treatment options vary with age, health, and the severity of the injury or wound.

SUMMARY

The present invention relates in part to methods of treating wounds, including burns. One objective of the present invention is to provide methods and materials for the treatment of traumatic injuries resulting from externally applied physical forces including surgical interventions, burns, and other wounds to promote repair and regeneration of injured tissue and to enhance the quality of life for surgery, trauma and burn patients.

In certain embodiments, the method includes the steps of contacting a wound with a treatment solution, and maintaining the treatment solution in the area of the wound for a period of time. In some embodiments, the treatment solution includes an ingredient selected from the group consisting of: an aqueous solution of electrolytes containing potassium ions at a concentration range of from about 35 to about 45 mM, sodium ions at a concentration range of from about 80 to about 120 mM, magnesium ions at a concentration range of from about 2 to about 10 mM, chloride ions at a concentration range of from about 15 to about 20 mM, and calcium ions at a concentration range of from about 0.01 to about 0.1 mM; an impermeant anion; mannitol; a macromolecular oncotic agent; at least one simple sugar; a substrate for the regeneration of ATP; a biological pH buffer effective under physiological hypothermic conditions; and combinations thereof.

In various embodiments, the treatment solution is HYPOTHERMOSOL®.

In various embodiments, the treatment solution includes glutathione, a vitamin E derivative, or combinations thereof.

In various embodiments, the treatment solution includes an antimicrobial agent, an antibacterial agent, an antifungal agent, an antiviral agent, or combinations thereof to mitigate against infection.

In various embodiments, the treatment solution is poured or otherwise directly contacted with the area of injury, so as to provide at least intermittent or transient duration of contact with the wound.

In various embodiments, the treatment solution is retained in the area of the wound.

In various embodiments, the method can include the step of irrigating the wound with the treatment solution to provide prolonged contact with the wound.

In various embodiments, the treatment solution includes an aerosol or mist that is sprayed on the wound or on a wound dressing.

In various embodiments, the temperature of the treatment solution is from -5 degrees Celsius to 25 degrees Celsius when applied to the wound.

In various embodiments, the temperature of the treatment in an ambient temperature.

In various embodiments, the treatment solution is in any of a frozen, partially frozen, wet ice, and/or liquid state or matter when applied to the wound.

In various embodiments, the method includes the step of providing a wound dressing and/or the step of treating a wound dressing with the treatment solution, before, during, or after application of the wound dressing to the wound. The method also can include the step of changing the wound dressing at least daily. In various embodiments, the method includes the step of providing the treatment solution.

In various embodiments, the treatment solution is combined with a gelling agent or additive to form a gelled or viscous treatment material to reduce migration of the treatment solution from the wound.

In various embodiments, the wound includes a burn, a traumatic injury, a medical intervention (e.g., a surgical incision), and combinations thereof.

In various embodiments, the treatment solution reduces scar formation and reduces wound healing time.

BRIEF DESCRIPTION OF DRAWINGS

The patent or application file contains at least one drawing executed in color. Copies of this patent or patent application publication with color drawings will be provided by the Office upon request and payment of the necessary fee.

The figures are not necessarily to scale, emphasis instead generally being placed upon illustrative principles. The figures are to be considered illustrative in all aspects and are not intended to limit the invention, the scope of which is defined only by the claims.

FIGS. 1A and IB are photographs of burn wounds treated with saline (FIG. 1A) and HYPOTHERMOSOL® (FIG. IB).

DETAILED DESCRIPTION

The present invention is based in part on the discovery that certain cell, tissue, and organ preservation media solution formulations, when utilized in a therapeutic regimen as a treatment solution as described herein, can provide a supportive environment conducive to promotion of recovery from wounds, including burns. More specifically, the present invention can be used to treat, ameliorate, manage and/or reduce the effects of acute and chronic wounds and other injuries resulting from, for example, disease, trauma, medical intervention, and burns. The present invention can be combined with traditional wound treatments as necessary.

As used herein, the term "wound" can refer to any natural, traumatic, medically-related, or burn-related injuries, and combinations of one or more of the foregoing. Wounds can be acute and/or chronic in nature. Natural injuries can include those injuries arising as a result of a disease condition or infection, including by way of non-limiting example, bacterial or viral infection and ulceration. Traumatic injuries can include any external stimulus that results in external or internal injury or damage to a subject (e.g., a human patient), including by way of non-limiting example, abrasions, scratches, lacerations, punctures, and thermal burn injuries from chemicals, radiation, or contact with any substance of sufficient temperature to cause thermal injury. Medically-related injuries can include changes to tissues, cells, organs or other body parts resulting from controlled purposeful medical interventions, including by way of non-limiting example, incisions, surgical manipulations, resections, sutures, and ablation.

The process of wound healing can be separated into several phases that overlap significantly: hemostasis, inflammation, proliferation, and remodeling. Hemostasis is the first response to wounding, whereby a clot formed from various clotting factors and initiation of the coagulation cascade stops blood loss from a damaged blood vessel. In addition, inflammatory cells are recruited to the site of the clot to destroy pathogens, remove cellular debris, and promote tissue healing and regrowth. Broughton et al., The Basic Science of Wound Healing, Plast. Reconstr. Surg., 117(7 Suppl): 12S-34S (2006).

During the inflammatory phase, blood supply and capillary permeability at the wound site are increased, thereby permitting more inflammatory cells and soluble factors to reach the wound and further promote pathogen destruction, removal of cellular debris, and healing. The development of inflammatory reactions is controlled by a number of complex biological pathways which include, for example, cytokines, complement, clotting, kinin, and fibrinolytic pathways by prostaglandins, leukotrienes, vasoactive mediators, basophils, and platelets. Kuby, Immunology, 664, 3rd ed., New York: W.H. Freeman and Company (1998).

The immediate inflammatory response is controlled in part by fast acting modulators, such as vasoactive amines and the products of the kinin system. Kuby, Immunology, 664 (3rd ed.; New York: W.H. Freeman and Company (1998)). Next, leukotrienes, recruit and activate other cells. Stvrtinova et al., Inflammation and Fever, Slovak Academy of Sciences, Slovakia: Academic Electronic Press (1995). Leukocytes also release factors that recruit and activate other cells. In response to more severe damage, mononuclear cells can infiltrate the damaged tissues to clear microbes and cellular debris. Kuby, Immunology, 664, 3rd ed., New York: W.H. Freeman and Company (1998).

During the proliferative phase of wound healing, epithelialization, angiogenesis, and ECM formation and remodeling occur. Macrophages and platelets release TGF-alpha and EGF which induce epithelial proliferation, and which also activate a cascade of other signaling events. Grotendorst et al, Journal of Cell Physiology, 139:617-23 (1989). For example, TNF-alpha and IL-1 induce fibroblast secretion of KGF-2 and IL-6, which stimulate keratinocyte migration, proliferation, and differentiation. Stojadinovic et al., Gynecol Oncol. 2008 Nov; 111(2 Suppl):S70-80. Epub 2008 Sep 14. Review. TNF-alpha and platelet-derived growth factor secretions induce the formation of new ECM and also recruit and activate fibroblasts to synthesize a matrix, which includes type III collagen, glycosaminoglycan, fibronectin, and integrin. Henry G, Garner WL. Inflammatory mediators in wound healing. Surg Clin North Am 2003;83:483-507.

An open wound bed eventually closes by myofibroblast-mediated wound contracture and migration of epithelial cells from the wound edge. Wound contracture reduces the size of the area needing repair. The dynamics of wound contracture are important, as reduced wound contraction can delay the healing process and protracted wound contraction can cause loss of tissue function and hypertrophic scarring. Stojadinovic et al, Gynecol Oncol. 2008 Nov; 111(2 Suppl):S70-80. Epub 2008 Sep 14. Review.

The maturation and remodeling phase of wound healing overlaps with the proliferative phase. The maturation and remodeling phase starts about eight days after wound infliction and can continue for a year or more. During maturation and remodeling, the matrix created during the proliferative phase is replaced by a more robust matrix comprised of type I collagen. Typically, a repaired wound fails to achieve the organization and strength of uninjured tissue. Broughton et al., The Basic Science of Wound Healing, Plast. Reconstr. Surg., 117(7 Suppl): 12S-34S (2006). An object of the present invention is to promote recovery from burns and wounds by the therapeutic application of a treatment solution comprised of ingredients and a formulation similar to that of intracellular-like cell, tissue, and organ preservation media.

In summary, it has been discovered that a treatment solution having a formulary similar to intracellular-like cell, tissue, or organ preservation media can promote healing of burns and wounds. A preferred treatment solution is HYPOTHERMOSOL® (BioLife Solutions, Inc., Bothell, WA), though it will be appreciated that other treatment solutions can be used as discussed herein. The therapeutic effects of the treatment solution are enabled by the incorporation of specific components in the formulation of the treatment solution, which work in combination and cumulatively to reduce cellular damage during the inflammatory phase, and by providing a nutrient environment during the proliferation and remodeling phases of recovery that supports maintenance and regrowth of the surrounding non-injured tissues. HYPOTHERMOSOL® is a sterile, protein-free, sera-free, GMP-grade preservation medium useful for hypothermic cell and tissue preservation, storage and transport. The treatment solutions and methods discussed herein can be used alone or in combination with other treatments.

Preferably, the treatment solution contemplated herein comprises an ingredient selected from the group consisting of: an aqueous solution of electrolytes containing potassium ions at a concentration range of from about 35 to about 45 mM, sodium ions at a concentration range of from about 80 to about 120 mM, magnesium ions at a concentration range of from about 2 to about 10 mM, chloride ions at a concentration range of from about 15 to about 20 mM, and calcium ions at a concentration range of from about 0.01 to about 0.1 mM; an impermeant anion; mannitol; a macromolecular oncotic agent; at least one simple sugar; a substrate for the regeneration of ATP; a biological pH buffer, and combinations thereof. In a preferred embodiment the treatment solution further includes glutathione, a vitamin E derivative, or combinations thereof.

There is a need, met by embodiments of the present invention, for improved healing of burns and wounds via the application of a treatment solution whose composition is similar to intracellular-like cell, tissue, or organ preservation media. Such a preferred treatment solution is HYPOTHERMOSOL®. Once HYPOTHERMOSOL® or a functional equivalent thereof is applied to the area of injury of a burn or wound, several components of the formulation provide various cellular benefits that promote recovery and healing from burns and wounds. An impermeant anion can be included that partially replaces chloride in the extracellular space and minimizes osmotic cell swelling. Specifically, and without wishing to be bound by theory, it is believed the impermeant anion fixes ions inside cells that are responsible for the oncotic pressure leading to osmotic cell swelling and eventual lysis. Lactobionate, another impermeant, is known to be a strong chelator of calcium and iron and may contribute to minimizing cell injury to due calcium influx and free radical generation. Sucrose and mannitol also can be included; the latter having properties of a hydroxl radical scavenger that reduces vascular resistance by inducing a prostaglandin mediated vasodilation, which is a key inflammatory phase response. Clinical grade dextran can be included as a colloid for oncotic support, to balance the hydrostatic pressure and help prevent interstitial edema. Dextran can improve the efficiency of the removal of erythrocytes from the tissue microvasculature by inhibiting red cell clumping, increasing intravascular osmotic pressure, and reducing vascular resistance. The ionic balance, notably, the Na⁺ / K⁺ and Ca²⁺ / Mg²⁺ ratios, is adjusted to restrict passive diffusional exchange when ionic pumps are shut down or at minimal activity levels. Adenosine is a multifaceted molecule and can be included as an essential substrate for the regeneration of ATP, and also as a vasoactive component to facilitate efficient vascular flushing by vasodilation. Glutathione can be included as a cellular antioxidant and hydroxl radical scavenger as a cofactor for glutathione peroxidase, which enables metabolism of lipid peroxides and hydrogen peroxide. Taylor et al., ASAIO Journal 40(3), M351-8 (1994).

As contemplated herein and described further elsewhere herein, a preferred treatment solution is protein-free and sera- free, and is adapted for cellular osmotic balance of tissue. One treatment solution is preferably HYPOTHERMOSOL® (BioLife Solutions, Inc., Bothell, WA).

It is understood that, when referenced throughout, HYPOTHERMOSOL® is identified and referenced as an exemplary treatment solution, and HYPOTHERMOSOL® is a preferred embodiment of treatment solution suitable for treating wounds with the materials and methods set forth herein. It is further understood that the present invention also contemplates functional equivalents of HYPOTHERMOSOL®; all that is required is that a treatment solution meet the functional requirements set forth herein and perform in a comparable manner when used in accordance with the present invention. Functional equivalents of HYPOTHERMOSOL® can be readily identified and recognized by the skilled practitioner practicing the teachings disclosed herein.

The HYPOTHERMOSOL® line of preservation solutions was initially designed to prepare and preserve cells, tissues and organs for hypothermic (i.e., low temperature, for example, about 2-10°C) environments and short-term hypothermic storage or transportation. For example, HYPOTHERMOSOL®-FRS (HTS-FRS) has been designed to address the molecular requirements of cells during a hypothermic (for example, about 2-10°C) preservation process. It has been formulated to reduce free radical accumulation in cells undergoing hypothermic preservation, which helps mediate the level of post-storage necrosis and apoptosis thereby directly reducing the level of cell death during and following the preservation interval. For example, HTS-FRS has been demonstrated to very effective at preserving myocardial and kidney tissues, both of which have high- energy demands that can lead to free radical accumulation.

However, it has been surprisingly and unexpectedly discovered that HYPOTHERMOSOL® and equivalent solutions also are useful as treatment solutions for treating wounds and facilitating the wound healing process. Such treatment solutions can be administered to the patient at temperatures ranging from -5 degrees Celsius to about 25 degrees Celsius or more. For example, the treatment solution can be about -5, about 0, about 5, about 10, about 15, about 20, or about 25 degrees Celsius when administered to a patient. In one embodiment, the treatment solution can be at a sub-freezing temperature, such as from about -5 degrees Celsius to about 0 degrees Celsius. In another embodiment, the treatment solution can be at hypothermic temperatures, such as from about 0 to about 18 degrees Celsius, from about 0 to about 15 degrees Celsius, from about 2 to about 18 degrees Celsius, and preferably from about 2 to about 12 degrees Celsius, and more preferably from about 2 to about 10 degrees Celsius, and even more preferably from about 2 to about 8 degrees Celsius. In yet another embodiment, the treatment solution can be at room temperature, such as from about 18 to about 25 degrees Celsius. In a further embodiment, the treatment solution can be at temperatures above 25 degrees Celsius. In one embodiment, the treatment solution is at ambient temperature when administered to the patient.

In some embodiments, the treatment solution comprises an ingredient selected from the group consisting of: an aqueous solution of electrolytes containing potassium ions at a concentration range of from about 35 to about 45 mM, sodium ions at a concentration range of from about 80 to about 120 mM, magnesium ions at a concentration range of from about 2 to about 10 mM, chloride ions at a concentration range of from about 15 to about 20 mM, and calcium ions at a concentration range of from about 0.01 to about 0.1 mM; an impermeant anion; mannitol; a macromolecular oncotic agent; at least one simple sugar; a substrate for the regeneration of ATP; a biological pH buffer effective under physiological hypothermic conditions; and combinations thereof. The treatment solution can further include glutathione, a vitamin E derivative, an antioxidant, or combinations thereof.

In various embodiments, the treatment solution is concentrated from about 1.1X to about 20X, such as, for example, about 1.1X, 1.5X, 2X, 3X, 4X, 5X, 10X, 15X, and 20X. For example, a 2X treatment solution comprises an ingredient selected from the group consisting of: an aqueous solution of electrolytes containing potassium ions at a concentration range of from about 70 to about 90 mM, sodium ions at a concentration range of from about 160 to about 240 mM, magnesium ions at a concentration range of from about 4 to about 20 mM, chloride ions at a concentration range of from about 30 to about 40 mM, and calcium ions at a concentration range of from about 0.02 to about 0.2 mM; an impermeant anion; mannitol; a macromolecular oncotic agent; at least one simple sugar; a substrate for the regeneration of ATP; a biological pH buffer effective under physiological hypothermic conditions; and combinations thereof. Concentrated treatment solutions can further include glutathione, a vitamin E derivative, an antioxidant, or combinations thereof. In another aspect, the invention provides a temperature range for a treatment solution, and/or the application of a treatment solution having temperature in the range of -5 degrees Celsius to 25 degrees Celsius when applied to the area of injury, so as to provide both an analgesic effect, as well as cell and tissue preservation effects, through the reduction in metabolism caused by exposure to a hypothermic environment, and by mitigating cell stress pathways and cellular and tissue responses present during the injury and healing phases of burns and wounds.

In another aspect, the invention provides for a treatment solution having a temperature range of -5 degrees Celsius to 25 degrees Celsius when applied to the area of injury, to be in, or transitioning between, any state of matter including but not limited to solid, slush, or liquid.

In another aspect, the invention provides a method of incorporating a treatment solution into various wound dressings and materials before application to the area of injury.

In yet another aspect, the present invention provides a method of utilizing a treatment solution by pouring or otherwise directly applying treatment solution with intermittent or transient contact with the area of injury.

In yet another aspect, the present invention provides a method for applying a treatment solution to the area of injury and containing the treatment solution on the area of injury by means of mechanical barriers that limit or eliminate solution runoff, so as to maintain more constant contact with the area of injury.

In yet another aspect, the present invention provides a method for continually circulating the treatment solution to irrigate the wound with treatment solution. Irrigation is known in the art and can be carried out using syringes, tubing, mechanical barriers, outflow orifices, and/or a pump, so as to provide more constant contact with the area of injury.

In yet another aspect, the present invention provides for an aerosolized embodiment of the treatment solution, which can be sprayed as a mist or aerosol on the area of injury.

In yet another aspect, the present invention includes the combination of a gelling agent or additive and the treatment solution, which forms a more viscous embodiment to enable more constant contact with the area of injury, as a result of less solution run-off. Gel components may include any of aloe, tea tree oil, Calendula, or any other biocompatible viscous material.

In yet another aspect, the present invention provides a treatment solution in a format and dosage suitable for emergency use, such as in a first aid kit or a military medical kit. By way of non-limiting example, the treatment solution could be packaged in individual doses and/or on a pre-soaked bandage.

In yet another aspect, the present invention includes any range of treatment frequency and/or duration as directed by a clinician. In addition, the treatment solution can be replaced or re-administered on the wound at any suitable interval, such as, for example, at least once every half hour, every hour, every two hours, every three hours, every four hours, every five hours, every six hours, every seven hours, every eight hours, every day, every other day, every three days, once a week, once a month and/or combinations and intermediates thereof.

The present invention is further illustrated by the following example, which is provided for illustration and not limitation.

EXAMPLE

Example 1 : in vivo treatment of wounds

An in vivo animal study has been initiated to confirm the efficacy of HYPOTHERMOSOL® in treating dermal wounds. The first data are included herein.

Three to four month old, 5 - 10 kg, female minipigs (Gottingen strain, Marshall BioResources, North Rose, NY) were acclimated for immunologic stability for at least five days prior to experimentation. This study was conducted according to a research proposal approved by the Institutional Animal Care and Use Committee (IACUC).

As shown in Table 1 , pigs were divided into groups for a burn wounding model. Briefly, animals were divided into three groups: no treatment (Group 1); treatment with saline, a commercially available wound treatment (Group 2); and treatment with HYPOTHERMOSOL® (Group 3). Animals were wounded and treatments were administered as described in more detail below. Wounds periodically were monitored and characterized over the time course of the study. Table 1. Summary of Study Design.

Wound induction: On Day 1 following acclimatization, each animal was suspended in a sling and anesthetized (ketamine/xylazine or isoflurane). Back hair was clipped and the back surgically scrubbed, and seven sites were marked on the back on both sides of the midline using a permanent marker. Individual sites were approximately 2.5 cm (1 inch) square, and were separated laterally by about 5 cm (2 inches) on each side. Partial thickness burns were created with an aluminum (square cross-section, 2.5-cm width) rod, which was pre -heated for 5 min in a boiling water bath and then placed in contact with the site (skin) with light pressure for 30 seconds. Following the induction of 7 burns per animal, pigs were recovered and returned to cages. Dry bandages were applied to protect the burns from any further self- inflicting injuries.

Treatment frequency: HYPOTHERMOSOL® and a commercially available wound treatment (saline solution) were administered twice per 24 hour period during dressing change to the animals, per Tables 1 and 2. Animal 151 received no treatment and served as a species negative control. Animals 251 and 252 were treated with commercial saline solution as follows: wound number 1 received no treatment and served as an in-subject negative control. Wound numbers 2, 3, and 4 were treated by soaking a thin gauze with 1 ml commercial saline solution and contacting the wound area momentarily to transfer the liquid to the wound, allowing the wound area to air dry, and then covering the wound area with a dry bandage. Wound numbers 5, 6, and 7 were treated by cutting a gauze bandage to fit the wound area, soaking this bandage in 1 ml commercial saline solution, applying the wet bandage to the wound, and covering the wet bandage with a dry bandage until the next dressing change and application. Animals 351 and 352 were treated with HYPOTHERMOSOL® as follows: wound number 1 received no treatment and served as an in-subject negative control. Wound numbers 2, 3, and 4 were treated by soaking a thin gauze with 1 ml HYPOTHERMOSOL® and contacting the wound area momentarily to transfer the liquid to the wound, allowing the wound area to air dry, and then covering the wound area with a dry bandage. Wound numbers 5, 6, and 7 were treated by cutting a gauze bandage to fit the wound area, soaking this bandage with 1 ml HYPOTHERMOSOL®, applying the wet bandage to the wound, and covering the wet bandage with a dry bandage until the next dressing change and application. HYPOTHERMOSOL® was applied at a temperature below room temperature, and saline solution control was applied at room temperature. Table 2. Wound Schematic and Drug Administration.

Observations: Animals were observed for 21 days and were then euthanized on day 22. Clinical observations and body weights were taken before experimentation and before euthanization. In addition, gross characterization included photographs, wound measurements, and Draize scoring (erythema), taken every two to three days to evaluate wound healing and recovery. Erythema, scabbing, and wound size are exemplary symptoms of burn wounds. Therefore, a reduction in and/or a rate of reduction in erythema, scabbing, and/or wound size are indicative of wound healing.

FIGS. 1A and B show exemplary images of burn wounds after 10 total treatments. FIG. 1A is a photograph of an exemplary wound from animal 251, wound site 3, which was treated with saline control solution as described above. FIG. IB is a photograph of an exemplary wound from animal 351, wound site 7, which was treated with a HYPOTHERMOSOL®-soaked bandage as described above.

It is expected that over time wounds treated with HYPOTHERMOSOL® will heal faster than wounds treated with saline, which is a common burn treatment. A visual difference is expected in the amount of redness and tissue damage between saline treated and HYPOTHERMOSOL® treated wounds. More specifically, with HYPOTHERMOSOL® treatment, more rapid reductions in wound size, inflammation, redness, edema, scabbing, and/or ulceration are expected.

In addition, after the last timepoint, gross necropsy is performed and wound and wound sites are harvested for histological evaluation. Tissues will be dehydrated, embedded in paraffin, sectioned at 3- to 5-μιη thicknesses, and stained, including with hematoxylin and eosin. Slides will be evaluated via light microscopy by a board-certified veterinary pathologist, including scoring of healing, inflammation, and fibrosis.

It also is expected that treatment with HYPOTHERMOSOL® will reduce scarring and/or scar tissue formation as compared to treatment with saline. Histologically, with HYPOTHERMOSOL® treatment, it is expected that the epithelial surface will be more intact with less blistering and loss of the epithelium. Surface ulceration and eschar also are expected to be reduced. In the subcutis, a reduction is expected in the amount of edema and vascular leakage. As will be appreciated, wounds can be treated with HYPOTHERMOSOL® for periods longer than 21 days, e.g., for as long as the patient is in need of such treatment. In the description, the invention is discussed in the context of burn wounds induced via contact with a heated aluminum surface; however, this embodiment is not intended to be limiting and those skilled in the art will appreciate that the invention also can be used for all types of wounds and burns.

The use of headings and sections in the application is not meant to limit the invention; each section can apply to any aspect, embodiment, or feature of the invention.

Throughout the application, where compositions are described as having, including, or comprising specific components, or where processes are described as having, including or comprising specific process steps, it is contemplated that compositions of the present invention also consist essentially of, or consist of, the recited components, and that the processes of the present invention also consist essentially of, or consist of, the recited process steps.

In the application, where an element or component is said to be included in and/or selected from a list of recited elements or components, it should be understood that the element or component can be any one of the recited elements or components and can be selected from a group consisting of two or more of the recited elements or components. Further, it should be understood that elements and/or features of a composition, an apparatus, or a method described herein can be combined in a variety of ways without departing from the spirit and scope of the present invention, whether explicit or implicit herein.

The use of the terms "include," "includes," "including," "have," "has," or "having" should be generally understood as open-ended and non-limiting unless specifically stated otherwise.

The use of the singular herein includes the plural (and vice versa) unless specifically stated otherwise. Moreover, the singular forms "a," "an," and "the" include plural forms unless the context clearly dictates otherwise. In addition, where the use of the term "about" is before a quantitative value, the present invention also includes the specific quantitative value itself, unless specifically stated otherwise. As used herein, the term "about" refers to a ±10% variation from the nominal value.

It should be understood that the order of steps or order for performing certain actions is immaterial so long as the present invention remains operable. Moreover, two or more steps or actions may be conducted simultaneously.

Where a range or list of values is provided, each intervening value between the upper and lower limits of that range or list of values is individually contemplated and is encompassed within the invention as if each value were specifically enumerated herein. In addition, smaller ranges between and including the upper and lower limits of a given range are contemplated and encompassed within the invention. The listing of exemplary values or ranges is not a disclaimer of other values or ranges between and including the upper and lower limits of a given range.

The invention may be embodied in other specific forms without departing from the spirit or essential characteristics thereof. The present aspects, embodiments, and features are therefore to be considered illustrative and not restrictive, the scope of the invention being indicated by the appended claims rather than by the foregoing description, and all changes, which come within the meaning and range of equivalency of the claims are therefore intended to be embraced therein.

What is claimed is:

Claims

1. A method of treating a wound, the method comprising:

contacting the wound with a treatment solution; and

maintaining the treatment solution in the area of the wound for a period of time,

wherein the treatment solution comprises an ingredient selected from the group consisting of: an aqueous solution of electrolytes containing potassium ions at a concentration range of from about 35 to about 45 mM, sodium ions at a concentration range of from about 80 to about 120 mM, magnesium ions at a concentration range of from about 2 to about 10 mM, chloride ions at a concentration range of from about 15 to about 20 mM, and calcium ions at a concentration range of from about 0.01 to about 0.1 mM; an impermeant anion; mannitol; a macromolecular oncotic agent; at least one simple sugar; a substrate for the regeneration of ATP; a biological pH buffer effective under physiological hypothermic conditions; and combinations thereof.

2. The method of claim 1, wherein the treatment solution is HYPOTHERMOSOL®.

3. The method of claim 1, wherein the treatment solution further comprises glutathione, a vitamin E derivative, or combinations thereof.

4. The method of claim 1 or 3, wherein the treatment solution is concentrated from about 1.1X to about 20X, such as, for example, about 1.1X, 1.5X, 2X, 3X, 4X,

5X, 10X, 15X, and 20X.

5. The method of claim 1 or 3, wherein the treatment solution further comprises an antimicrobial agent, an antibacterial agent, an antifungal agent, an antiviral agent, or combinations thereof.

6. The method of claim 1, wherein the treatment solution is poured or otherwise directly contacted with the area of injury, so as to provide at least intermittent or transient contact with the wound.

7. The method of claim 1, wherein the treatment solution is retained in the area of the wound.

8. The method of claim 1, comprising the step of irrigating the wound with treatment solution to provide prolonged contact with the wound.

9. The method of claim 1, wherein the treatment solution comprises an aerosol or mist that is sprayed on the wound.

10. The method of claim 1, wherein the temperature of the treatment solution is from about -5 degrees Celsius to about 25 degrees Celsius when applied to the wound.

11. The method of claim 1 , wherein the temperature of the treatment solution is an ambient temperature.

12. The method of claim 1, wherein the treatment solution is in a frozen, partially frozen, wet ice, and/or liquid state when applied to the wound.

13. The method of claim 1, comprising the step of providing a wound dressing.

14. The method of claim 1, comprising the step of treating a wound dressing with treatment solution, before, during, or after application of the wound dressing to the wound.

15. The method of claim 12 or 13, comprising the step of changing the wound dressing at least daily.

16. The method of claim 1, comprising the step of providing the treatment solution.

17. The method of claim 1, wherein the treatment solution is combined with a gelling agent or additive to form a gelled or viscous treatment material to reduce migration of the treatment solution from the wound.

18. The method of claim 1 , wherein the wound comprises a burn.

19. The method of claim 1, wherein the wound comprises a traumatic injury.

20. The method of claim 1, wherein the wound comprises a medical intervention.

21. The method of claim 1, wherein the treatment solution reduces scar formation.

22. The method of claim 1, wherein the treatment solution reduces wound healing time.