Patent Application
20220347340
The present technology relates generally to wound dressing compositions including a mixture of a collagen and/or chitosan and oxidized regenerated cellulose (ORC) and including a layer disposed within the composition and adjacent to an external surface that includes citric acid. Methods of manufacture as well as kits regarding the same are also provided.
The following description of the background of the present technology is provided simply as an aid in understanding the present technology and is not admitted to describe or constitute prior art to the present technology.
Infections can retard wound healing and, if untreated, can result in tissue loss, systemic infections, septic shock, and death. Moreover, in addition to vegetative or free-floating bacteria present in a wound, bacterial biofilms may also form in a wound presenting further challenges in wound therapy, particularly chronic wounds. A biofilm is an association of microorganisms that can adhere to a surface forming three-dimensional microbial communities. The ability of bacteria to form these complex biofilms can impede a host's defense mechanisms against pathogens. Antimicrobials have been used to attempt to prevent, reduce, inhibit, and/or disrupt biofilm formation in wounds.
In an aspect, a wound dressing composition that includes a first external surface and a second external surface is provided. The wound dressing composition includes about 30 wt. % to about 95 wt. % of one or more of collagen and chitosan; and about 30 wt. % to about 70 wt. % oxidized regenerated cellulose (ORC). A first layer disposed within the wound dressing composition adjacent to the first external surface further includes citric acid.
In a related aspect, a method of manufacturing a wound dressing composition of any embodiment of the present technology from a dressing composition that includes a first exterior surface and a second exterior surface is provided, where the method includes applying a first citric acid solution to the first exterior surface of the dressing composition. The first citric acidic solution includes a first solvent and about 50 mM to about 400 mM citric acid. In another aspect, a wound dressing composition prepared according to the methods described herein in any embodiment of the present technology is provided.
In a further related aspect, a kit is provided that includes a wound dressing composition of any embodiment described herein as well as instructions for use.
In another related aspect, a method for providing therapy to a tissue site that includes positioning a wound dressing composition of any embodiment of the present technology adjacent to the tissue site is provided. In any embodiment herein, the method for providing therapy may include providing negative pressure therapy to the tissue site.
In yet another related aspect, a method of reducing or preventing the formation of a biofilm in a tissue site that includes applying a wound dressing composition of any embodiment of the present technology to the tissue site is provided.
It is to be appreciated that certain aspects, modes, embodiments, variations and features of the present methods are described below in various levels of detail in order to provide a substantial understanding of the present technology.
The definitions of certain terms as used in this specification are provided below. Unless defined otherwise, all technical and scientific terms used herein generally have the same meaning as commonly understood by one of ordinary skill in the art to which this present technology belongs.
The following terms are used throughout as defined below.
As used herein and in the appended claims, singular articles such as “a”, “an”, and “the” and similar referents in the context of describing the elements (especially in the context of the following claims) are to be construed to cover both the singular and the plural, unless otherwise indicated herein or clearly contradicted by context. Recitation of ranges of values herein are merely intended to serve as a shorthand method of referring individually to each separate value falling within the range, unless otherwise indicated herein, and each separate value is incorporated into the specification as if it were individually recited herein. All methods described herein can be performed in any suitable order unless otherwise indicated herein or otherwise clearly contradicted by context. The use of any and all examples, or exemplary language (e.g., “such as”) provided herein, is intended merely to better illuminate the embodiments and does not pose a limitation on the scope of the claims unless otherwise stated. No language in the specification should be construed as indicating any non-claimed element as essential.
As used herein, “about” will be understood by persons of ordinary skill in the art and will vary to some extent depending upon the context in which it is used. If there are uses of the term which are not clear to persons of ordinary skill in the art, given the context in which it is used, “about” will mean up to plus or minus 10% of the particular term—for example, “about 10 wt. %” would mean “9 wt. % to 11 wt. %.” It is to be understood that when “about” precedes a term, the term is to be construed as disclosing “about” the term as well as the term without modification by “about”—for example, “about 10 wt. %” discloses “9 wt. % to 11 wt. %” as well as discloses “10 wt. %.”
As understood by one of ordinary skill in the art, “molecular weight” (also known as “relative molar mass”) is a dimensionless quantity but is converted to molar mass by multiplying by 1 gram/mole—for example, collagen with a weight-average molecular weight of 5,000 has a weight-average molar mass of 5,000 g/mol.
The term “substantially free” as used herein means the indicated component is present in an amount less than about 0.001 wt. % weight percent (wt. %) of the composition.
As used herein, the term “biofilm” refers to an association of microorganisms, e.g., single or multiple species, that can be encased or embedded in a matrix material, which may be self-produced by resident microorganisms. The biofilm may be present or adhere to living and/or non-living surfaces, e.g., tissue, a wound, medical implants, such as but not limited to orthopedic implants, dental implants, catheters, stents and so on. Exemplary microorganisms include, but are not limited to bacteria, e.g., Gram-negative bacteria, such as Pseudomonas aeruginosa, Gram-positive bacteria, such as Staphylococcus aureus and Streptococcus mutans, and fungi, such as yeasts, e.g., Candida albicans. The term “matrix material” is intended to encompass extracellular polymeric substances. Exemplary matrix materials include, but are not limited to polysaccharides, glycoproteins and/or nucleic acids. The term “biofilm” is further intended to include biological films that develop and persist at interfaces in aqueous environments. The language “biofilm development” or “biofilm formation” is intended to include the formation, growth, and modification of the bacterial colonies contained with biofilm structures, as well as the synthesis and maintenance of the exopolysaccharide of the biofilm structures. “Reducing” or “disrupting” a biofilm includes reducing the number of total viable microorganisms making up at least part of the biofilm, for example, as measured by total viable counts (TVC) of microorganisms (e.g., bacteria, yeast).
As used herein, the “administration” of a wound dressing composition to a subject includes any route of introducing or delivering to a subject a diagnostic wound dressing composition to perform its intended function. Administration can be carried out by any suitable route, including but not limited to, topical administration. Administration includes self-administration and the administration by another.
As used herein, the terms “individual”, “patient”, or “subject” can be an individual organism, a vertebrate, a mammal, or a human. In some embodiments, the individual, patient, or subject is a human.
“Treating” or “treatment” as used herein covers the treatment of a wound described herein, in a subject, such as a human, and includes: (i) inhibiting a wound, i.e., arresting its development; (ii) relieving a wound, i.e., causing regression of the wound; (iii) slowing progression of the wound; and/or (iv) inhibiting, relieving, or slowing progression of one or more symptoms of the wound. In some embodiments, treatment means that the symptoms associated with the wound are alleviated, reduced, cured, or placed in a state of remission.
“Tissue site” as used herein refers to a wound, defect, or other treatment target located on or within tissue, including but not limited to, bone tissue, adipose tissue, muscle tissue, neural tissue, dermal tissue, vascular tissue, connective tissue, cartilage, tendons, or ligaments. A wound may include chronic, acute, traumatic, subacute, and dehisced wounds, partial-thickness burns, ulcers (such as diabetic, pressure, or venous insufficiency ulcers), flaps, grafts, or a combination of any two or more thereof. The term “tissue site” may also refer to areas of any tissue that are not necessarily wounded or defective, but are instead areas in which it may be desirable to add or promote the growth of additional tissue.
It is also to be appreciated that the various modes of treatment of wounds as described herein are intended to mean “substantial,” which includes total but also less than total treatment, and wherein some biologically or medically relevant result is achieved. The treatment may be a continuous prolonged treatment for a chronic wound or a single administration, or a few administrations for the treatment of an acute wound.
A wound can become infected by microbes. An infected wound is a wound in which bacteria or other microorganisms have colonized, causing a deterioration and delay in the healing of the wound. Thus, a reduction in bacterial colonization is vital in wound therapy.
A biofilm is an association of microorganisms (e.g., single or multiple species) that can adhere to a surface forming three-dimensional microbial communities, which can have coordinated multi-cellular behavior. Typically, biofilms can produce extracellular polysaccharides thereby forming an extracellular matrix in which the bacteria are embedded. The ability of bacteria to form these complex biofilms can impede a host's defense mechanisms against pathogens. As such, biofilms often display a heightened tolerance to antimicrobial treatment.
The present disclosure is directed to wound dressing compositions that include at least one layer of citric acid disposed within the wound dressing composition, which may prevent, reduce, inhibit, or disrupt biofilm levels in a wound upon application, and over time.
Thus, in an aspect, a wound dressing composition is provided that includes a first external surface and a second external surface. The wound dressing composition includes about 30 wt. % to about 95 wt. % of one or more of collagen and chitosan, and about 30 wt. % to about 70 wt. % oxidized regenerated cellulose (ORC). In any embodiment herein, the wound dressing composition may include a weight ratio of the one or more of collagen and chitosan to ORC from about 70:30 to about 30:70. The weight ratio of the one or more of collagen and chitosan to ORC may be about 70:30, about 69:31, about 68:32, about 67:33, about 66:34, about 65:35, about 64:36, about 63:37, about 62:38, about 61:39, about 60:40, about 59:41, about 58:42, about 57:43, about 56:44, about 55:45, about 54:46, about 53:47, about 52:48, about 51:49, about 50:50, about 49:51, about 48:52, about 47:53, about 46:54, about 45:55, about 44:56, about 43:57, about 42:58, about 41:59, about 40:60, about 39:61, about 38:62, about 37:63, about 36:64, about 35:65, about 34:66, about 32:68, about 31:69, about 30:70, or any range including and/or in between any two of these values.
A first layer that includes citric acid is disposed within the wound dressing composition adjacent to the first external surface of the wound dressing composition. In any embodiment disclosed herein, the first layer that includes citric acid may extend inward towards the interior of the wound dressing composition. In any embodiment disclosed herein, the wound dressing composition may also include a second layer disposed within the wound dressing composition adjacent to the second external surface, wherein the second layer independently includes citric acid. The first layer and (when present) second layer may each independently have an average thickness of about 10 μm to about 3,500 μm; thus, the first layer and (when present) second layer may each independently have an average thickness of about 10 μm, about 20 μm, about 30 μm, about 40 μm, about 50 μm, about 60 μm, about 70 μm, about 80 μm, about 90 μm, about 100 μm, about 150 μm, about 200 μm, about 250 μm, about 300 μm, about 350 μm, about 400 μm, about 450 μm, about 500 μm, about 550 μm, about 600 μm, about 650 μm, about 700 μm, about 750 μm, about 800 μm, about 850 μm, about 900 μm, about 950 μm, about 1,000 μm, about 1,250 μm, about 1,500 μm, about 1,750 μm, about 2,000 μm, about 2,250 μm, about 2,500 μm, about 2,750 μm, about 3,000 μm, about 3,250 μm, about 3,500 μm, or any range including an/or in between any two of these values. The average thickness may be determined by using a calibrated optical microscope to measure the citric acid-containing band of a cross-section of the wound dressing composition. While a variety of methods of determining the average thickness of a layer may be utilized, one approach includes measuring the layer thickness of ten (10) equidistant points of a cross-section of the wound dressing composition and calculating the average from the ten measurements. Citric acid may be included in the first layer and (when present) the second layer at the same or differing amounts. The amount of citric acid included in the first layer and (when present) the second layer may independently for each layer be about 0.001 wt. % to about 20 wt. % based on the total weight of the wound dressing composition; thus, the amount of citric acid included in the first layer and (when present) the second layer may independently for each layer be (by total weight of the wound dressing composition) about 0.001 wt. %, about 0.002 wt. %, about 0.003 wt. %, about 0.004 wt. %, about 0.005 wt. %, about 0.006 wt. %, about 0.007 wt. %, about 0.008 wt. %, about 0.009 wt. %, about 0.01 wt. %, about 0.02 wt. %, about 0.03 wt. %, about 0.04 wt. %, about 0.05 wt. %, about 0.06 wt. %, about 0.07 wt. %, about 0.08 wt. %, about 0.09 wt. %, about 0.1 wt. %, about 0.2 wt. %, about 0.3 wt. %, about 0.4 wt. %, about 0.5 wt. %, about 0.6 wt. %, about 0.7 wt. %, about 0.8 wt. %, about 0.9 wt. %, about 1 wt. %, about 2 wt. %, about 3 wt. %, about 4 wt. %, about 5 wt. %, about 0.6 wt. %, about 0.7 wt. %, about 0.8 wt. %, about 0.9 wt. %, about 1 wt. %, about 2 wt. %, about 3 wt. %, about 4 wt. %, about 5 wt. %, about 6 wt. %, about 7 wt. %, about 8 wt. %, about 9 wt. %, about 10 wt. %, about 11 wt. %, about 12 wt. %, about 13 wt. %, about 14 wt. %, about 15 wt. %, about 16 wt. %, about 17 wt. %, about 18 wt. %, about 19 wt. %, about 20 wt. %, or any range including and/or in between any two of these values.
While the first layer and (when present) the second layer each include citric acid, the remaining wound dressing composition may be substantially free of citric acid. In any embodiment herein, the amount of citric acid in the remaining wound dressing composition (i.e., not including the first layer and (when present) the second layer) may be less than about 0.0001 wt. %. However, in any embodiment herein, it may be that the first layer extends the entire thickness of the wound dressing composition (where there would be no second layer) and there is a gradient of concentration of the citric acid between the first external surface and the second external surface. The wound dressing composition may have an average thickness of about 0.01 cm to about 0.5 cm; thus, the thickness of the wound dressing composition may be about 0.01 cm, about 0.02 cm, about 0.03 cm, about 0.04 cm, about 0.05 cm, about 0.06 cm, about 0.07 cm, about 0.08 cm, about 0.09 cm, about 0.10 cm, about 0.15 cm, about 0.20 cm, about 0.25 cm, about 0.30 cm, about 0.35 cm, about 0.40 cm, about 0.45 cm, about 0.50 cm, or any range including and/or in between any two of these values
The first layer and (when present) the second layer may each independently include a dye marker. The dye marker of the first and/or second layer may independently for each layer be E124, fluorescein isothiocyanate (FITC), 4′,6-diamidino-2-phenylindole (DAPI), methylene blue, erythrosine B, ponceau S, alura red, SYBR green, alcian blue, brilliant blue G, calcein blue, cardio green, crystal violet, nile blue, india ink, brilliant blue, indigo carmine, sudan III, methyl green, oil red, pyronin Y, tattoo ink, purpurin, phloxine B, picric acid, carbon nanotubes, a fuchsin, resazurin, a trichrome, or a combination of any two or more thereof. For example, the first and/or second layer may independently include a mixture of FITC, erythrosine B, ponceau S, brilliant blue and cardio green. As another example, the first and/or second layer may independently include a mixture of methylene blue, SYBR green, and purpurin. Yet another example is where the first and/or second layer independently include a mixture of oil red, nile blue and sudan III. In any embodiment herein, it may be that the first external surface is a wound-facing external surface. In any embodiment herein, it may be the second external surface is an environmental-facing external surface.
In any embodiment disclosed herein, the collagen of the wound dressing composition may include a mammalian collagen, such as a bovine collagen, a human collagen, or a combination thereof. The bovine collagen may include one or both of bovine collagen type I and bovine collagen type III. Exemplary types of human collagen include, but are not limited to, human collagen type I, human collagen type II, human collagen type III, human collagen type IV, human collagen type V, human collagen type VI, human collagen type VII, human collagen type VIII, human collagen type IX, human collagen type X, human collagen type XI, human collagen type XII, human collagen type XIII, human collagen type XIV, human collagen type XV, human collagen type XVI, human collagen type XVII, human collagen type XVIII, human collagen type XIX, human collagen type XX, human collagen type XXI, human collagen type XXII, human collagen type XXIII, human collagen type XXIV, human collagen type XXV, human collagen type XXVI, human collagen type XXVII, as well as a combination of any two or more thereof. The collagen of any embodiment herein may be a Type I collagen, a Type II collagen, a Type III collagen, may be obtained from any natural source, may be chemically-modified collagen (e.g., an atelocollagen obtained by removing the immunogenic telopeptides from natural collagen), or may be a combination of any two or more thereof. For example, the collagen may include collagen obtained from bovine corium that has been rendered largely free of non-collagenous components, for example, including fat, non-collagenous proteins, polysaccharides, and other carbohydrates, such as by procedures described in U.S. Pat. Nos. 4,614,794 and 4,320,201, each of which is incorporated herein by reference. In any embodiment disclosed herein, the wound dressing composition may include a weight ratio of human collagen type I to human collagen type III of about 100:0, about 90:10, about 80:20, about 70:30, about 60:40, about 50:50, about 40:60, about 30:70, about 20:80, about 10:90, about 0:100, or any range including and/or in between any two of these values. The ratio by weight of human collagen type I to human collagen type III may be greater than about 50:50, or greater than about 70:30. The collagen of any embodiment herein may include a weight ratio of type I bovine collagen to type III bovine collagen of about 85:15.
As discussed above, the wound dressing composition includes about 30 wt. % to about 95 wt. % of one or more of collagen and chitosan. Thus, the amount of collagen and/or chitosan in the wound dressing composition may be about 30 wt. %, about 32 wt. %, about 34 wt. %, about 36 wt. %, about 38 wt. %, about 40 wt. %, about 42 wt. %, about 44 wt. %, about 46 wt. %, about 48 wt. %, about 50 wt. %, about 52 wt. %, about 54 wt. %, about 56 wt. %, about 58 wt. %, about 60 wt. %, about 62 wt. %, about 64 wt. %, about 66 wt. %, about 68 wt. %, about 70 wt. %, about 72 wt. %, about 74 wt. %, about 76 wt. %, about 78 wt. %, about 80 wt. %, about 82 wt. %, about 84 wt. %, about 86 wt. %, about 88 wt. %, about 90 wt. %, about 92 wt. %, about 94 wt. %, about 95 wt. %, or any range including and/or in between any two of these values. The collagen included in the wound dressing composition may have a weight-average molecular weight of about 5,000 to about 100,000. For example, in any embodiment disclosed herein, the collagen may have a weight-average molecular weight of about 5,000, about 6,000, about 7,000, about 8,000, about 9,000, about 10,000, about 12,000, about 14,000, about 16,000, about 18,000, about 20,000, about 22,000, about 24,000, about 26,000, about 28,000, about 30,000, about 32,000, about 34,000, about 36,000, about 38,000, about 40,000, about 45,000, about 50,000, about 55,000, about 60,000, about 65,000, about 70,000, about 75,000, about 80,000, about 85,000, about 90,000, about 95,000, about 100,000, or any range including and/or in between any two of these values. The chitosan of the wound dressing composition may have a weight-average molecular weight of about 100 to about 220,000; thus, in any embodiment herein, the chitosan of the wound dressing may have a weight-average molecular weight of about 100, about 200, about 300, about 400, about 500, about 600, about 700, about 800, about 900, about 1,000, about 2,000, about 3,000, about 4,000, about 5,000, about 6,000, about 7,000, about 8,000, about 9,000, about 10,000, about 12,000, about 14,000, about 16,000, about 18,000, about 20,000, about 22,000, about 24,000, about 26,000, about 28,000, about 30,000, about 32,000, about 34,000, about 36,000, about 38,000, about 40,000, about 45,000, about 50,000, about 55,000, about 60,000, about 65,000, about 70,000, about 75,000, about 80,000, about 85,000, about 90,000, about 95,000, about 100,000, about 120,000, about 140,000, about 160,000, about 180,000, about 200,000, about 220,000, or any range including and/or in between any two of these values.
Oxidized regenerated cellulose (ORC) may be produced by the oxidation of cellulose, for example with dinitrogen tetroxide and/or as described in U.S. Pat. No. 3,122,479 (incorporated herein by reference). Not intending to be bound by theory, it is believed that this process may convert primary alcohol groups on the saccharide residues of the cellulose to carboxylic acid groups, for example, forming uronic acid residues within the cellulose chain. The oxidation may not proceed with complete selectivity, and as a result hydroxyl groups on carbons 2 and 3 of the saccharide residue may be converted to the keto form. These ketone units may introduce an alkali labile link, which at pH 7 or higher initiates the decomposition of the polymer via formation of a lactone and sugar ring cleavage. As a result, oxidized regenerated cellulose is biodegradable and bioresorbable under physiological conditions. ORC is available with a variety of degrees of oxidation and hence rates of degradation. The ORC may include particles, fibers, or both; in any embodiment disclosed herein, the ORC may be in the form of particles, such as fiber particles or powder particles. In embodiments that include ORC fibers, the ORC fibers may have a volume fraction such that at least 80% of the fibers have lengths in the range from about 5 μm to about 1000 μm, or from about 250 μm to about 450 μm.
As discussed above, the ORC of the wound dressing composition may include about 30 wt. % to about 70 wt. % based on the weight of the wound dressing composition of the ORC. In any embodiment disclosed herein, the ORC may have a weight-average molecular weight of about 50,000 to about 1,000,000. Thus, the wound dressing composition may include an ORC of any embodiment disclosed herein in an amount of about 30 wt. %, about 32 wt. %, about 34 wt. %, about 36 wt. %, about 38 wt. %, about 40 wt. %, about 42 wt. %, about 44 wt. %, about 46 wt. %, about 48 wt. %, about 50 wt. %, about 52 wt. %, about 54 wt. %, about 56 wt. %, about 58 wt. %, about 60 wt. %, about 62 wt. %, about 64 wt. %, about 66 wt. %, about 68 wt. %, about 70 wt. %, or any range including and/or in between any two of these values based on the weight of the wound dressing composition. The ORC may have a weight-average molecular weight of about 50,000, about 100,000, about 150,000, about 200,000, about 250,000, about 300,000, about 350,000, about 400,000, about 450,000, about 500,000, about 550,000, about 600,000, about 650,000, about 700,000, about 750,000, about 800,000, about 850,000, about 900,000, about 950,000, about 1,000,000, or any range including and/or in between any two of these values.
In any embodiment disclosed herein, the wound dressing composition may include about 0.001 wt. % to about 5 wt. % of an antimicrobial agent. Thus, the total amount of antimicrobial agent(s) in the wound dressing composition may be about 0.001 wt. %, about 0.002 wt. %, about 0.003 wt. %, about 0.004 wt. %, about 0.005 wt. %, about 0.006 wt. %, about 0.007 wt. %, about 0.008 wt. %, about 0.009 wt. %, about 0.01 wt. %, about 0.02 wt. %, about 0.03 wt. %, about 0.04 wt. %, about 0.05 wt. %, about 0.06 wt. %, about 0.07 wt. %, about 0.08 wt. %, about 0.09 wt. %, about 0.1 wt. %, about 0.2 wt. %, about 0.3 wt. %, about 0.4 wt. %, about 0.5 wt. %, about 0.6 wt. %, about 0.7 wt. %, about 0.8 wt. %, about 0.9 wt. %, about 1 wt. %, about 2 wt. %, about 3 wt. %, about 4 wt. %, about 5 wt. %, or any range including and/or in between any two of these values. The antimicrobial agent may include a penicillin, a streptomycin, ionic silver, chlorhexidine, a poly(hexamethylene biguanide) (PHMB), iodine, or a combination of any two or more thereof. Ionic silver may be provided in a variety of forms including as pharmaceutically acceptable salts, where representative examples include but are not limited to silver oxide, silver oxysalts, silver chromate, silver allantoinate, silver borate, silver glycerolate, silver nitrate, silver acetate, silver chloride, silver sulfate, silver lactate, silver bromide, silver iodide, silver carbonate, silver citrate, silver laurate, silver deoxycholate, silver salicylate, silver p-aminobenzoate, silver p-aminosalicylate, or a combination of any two or more thereof. In addition, while ionic silver may initially be incorporated into the wound dressing composition as a silver salt, at least a portion (if not substantially all) of the ionic silver may be complexed to the ORC in the wound dressing composition.
In any embodiment disclosed herein, the wound dressing composition may include one or more additional biomaterials such as gelatin, fibronectin, hyaluronic acid, polysaccharides, and a combination of any two or more thereof. The total amount of the one or more additional biomaterials in the wound dressing composition may be about 1 wt. %, about 2 wt. %, about 3 wt. %, about 4 wt. %, about 5 wt. %, about 6 wt. %, about 7 wt. %, about 8 wt. %, about 9 wt. %, about 10 wt. %, about 11 wt. %, about 12 wt. %, about 13 wt. %, about 14 wt. %, about 15 wt. %, about 16 wt. %, about 17 wt. %, about 18 wt. %, about 19 wt. %, about 20 wt. %, about 21 wt. %, about 22 wt. %, about 23 wt. %, about 24 wt. %, about 25 wt. %, or any range including and/or in between any two of these values, based on the weight of the wound dressing composition.
In any embodiment disclosed herein, in addition to citric acid the wound dressing composition may include about 0.001 wt. % to about 5 wt. % of one or more antioxidants. Thus, the total amount of antioxidants may be about 0.001 wt. %, about 0.002 wt. %, about 0.003 wt. %, about 0.004 wt. %, about 0.005 wt. %, about 0.006 wt. %, about 0.007 wt. %, about 0.008 wt. %, about 0.009 wt. %, about 0.01 wt. %, about 0.02 wt. %, about 0.03 wt. %, about 0.04 wt. %, about 0.05 wt. %, about 0.06 wt. %, about 0.07 wt. %, about 0.08 wt. %, about 0.09 wt. %, about 0.1 wt. %, about 0.2 wt. %, about 0.3 wt. %, about 0.4 wt. %, about 0.5 wt. %, about 0.6 wt. %, about 0.7 wt. %, about 0.8 wt. %, about 0.9 wt. %, about 1 wt. %, about 2 wt. %, about 3 wt. %, about 4 wt. %, about 5 wt. %, or any range including and/or in between any two of these values. Exemplary antioxidants include, but are not limited to, anthocyanins, astaxanthin, bilirubin, canthaxanthin, capsaicin, curcumin, coenzyme Q10, eugenol, flavanol, flavonolignans, flavanone, flavone, flavonol, iodide, isoflavone phytoestrogen, lutein, lycopene, manganese, melatonin, N-acetylcysteine, oxalic acid, phenolic acid, phytic acid, R-α-lipoic acid, stilbenoid, tocopherol, tocotrienol, vitamin A, vitamin C, vitamin E, xanthones, zeaxanthin, α-carotene, β-carotene, as well as combinations of any two or more thereof.
Examples of anthocyanins include, but are not limited to cyanidin, delphinidin, malvidin, pelargonidin, peonidin, petunidin, as well as combinations of any two or more combination thereof.
Examples of flavanols include, but are not limited to catechin, epicatechin, theaflavin, thearubigins, gallocatechin, epigallocatechin, or any gallate ester thereof, as well as combinations of any two or more thereof.
Examples of flavanones include, but are not limited to eriodictyol, hesperetin, naringenin, and combinations of any two or more thereof.
Examples of flavones include, but are not limited to apigenin, luteolin, tangeritin, and combinations of any two or more thereof.
Examples of flavonols include, but are not limited to isorhamnetin, kaempferol, myricetin, proanthocyanidins, quercetin, rutin, and combinations of any two or more thereof.
Examples of isoflavone phytoestrogens include, but are not limited to daidzein, genistein, glycitein, and combinations of any two or more thereof.
Examples of phenolic acids include, but are not limited to chicoric acid, chlorogenic acid, cinnamic acid, ellagic acid, ellagitannins, gallic acid, gallotannins, rosmarinic acid, salicylic acid, or any ester thereof, and combinations of any two or more thereof.
Examples of stillbenoids includes, but is not limited to resveratrol, pterostilbene, and mixtures thereof.
In any embodiment disclosed herein, the first layer and (when present) the second layer that includes citric acid may further include one or more binders.
In any embodiment disclosed herein, the wound dressing composition of the present technology may be capable of preventing, reducing, inhibiting, or disrupting biofilm formation in a wound. Reducing a biofilm includes reducing the number of total viable microorganisms making up at least part of the biofilm, for example, as measured by total viable counts (TVC) of microorganisms (e.g., bacteria, yeast). The biofilm may include bacteria such as Pseudomonas aeruginosa, Staphylococcus aureus and Streptococcus mutans. The biofilm may also include fungi including, but not limited to, yeasts (such as Candida albicans). The wound dressing composition of the present technology may be capable of preventing, reducing, inhibiting, and/or disrupting a biofilm in a wound by ≥about 10% to about 100%, after 24 hours in vitro or in vivo exposure. For example, the wound dressing composition of the present technology may be capable of preventing, reducing, inhibiting, and/or disrupting a biofilm in a wound by about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 99%, about 100%, or any range including and/or in between any two of these values.
The therapeutic efficacy of the wound dressing composition of the present technology can be assayed using any method known to those in the art. An exemplary method to test the therapeutic efficacy of the wound dressing composition of the present technology is the colony drip flow reactor (C-DFR) assay (see Lipp, C., et al., J. Wound Care, 19:220-226(2010), which is incorporated herein by reference).
In any embodiment disclosed herein, the wound dressing composition of the present disclosure may be sterile. In any embodiment disclosed herein, the wound dressing composition of the present disclosure may be packaged in a microorganism-impermeable container.
In a related aspect, a method of manufacturing a wound dressing composition of any embodiment of the present technology from a dressing composition that includes a first exterior surface and a second exterior surface is provided, where the method includes applying a first citric acid solution to the first exterior surface of the dressing composition. The first citric acidic solution includes a first solvent and about 50 mM to about 400 mM citric acid. The first citric acid solution may be applied at a volume (determined at 20° C.) of about 1 microliter (μL) to about 50 μL per square centimeter of the first exterior surface of the dressing composition. Thus, the amount of first citric acid solution applied may be about 1 μL, about 2 μL, about 3 μL, about 4 μL, about 5 μL, about 6 μL, about 7 μL, about 8 μL, about 9 μL, about 10 μL, about 11 μL, about 12 μL, about 13 μL, about 14 μL, about 15 μL, about 16 μL, about 17 μL, about 18 μL, about 19 μL, about 20 μL, about 21 μL, about 22 μL, about 23 μL, about 24 μL, about 25 μL, about 26 μL, about 27 μL, about 28 μL, about 29 μL, about 30 μL, about 32 μL, about 34 μL, about 36 μL, about 38 μL, about 40 μL, about 42 μL, about 44 μL, about 46 μL, about 48 μL, about 50 μL, or any range including and/or in between any two of these values. Such application may (but is not required to) occur at a temperature of about 20° C. to about 25° C.
In any embodiment herein, the method may or may not further include applying a second citric acid solution (including a second solvent and about 50 mM to about 400 mM citric acid) to the second exterior surface of the dressing composition. The second citric acid solution may be applied at a volume (determined at 20° C.) of about 1 μL to about 50 μL per square centimeter of the second exterior surface of the dressing composition. Thus, the amount of second citric acid solution applied may be about 1 μL, about 2 μL, about 3 μL, about 4 μL, about 5 μL, about 6 μL, about 7 μL, about 8 μL, about 9 μL, about 10 μL, about 11 μL, about 12 μL, about 13 μL, about 14 μL, about 15 μL, about 16 μL, about 17 μL, about 18 μL, about 19 μL, about 20 μL, about 21 μL, about 22 μL, about 23 μL, about 24 μL, about 25 μL, about 26 μL, about 27 μL, about 28 μL, about 29 μL, about 30 μL, about 32 μL, about 34 μL, about 36 μL, about 38 μL, about 40 μL, about 42 μL, about 44 μL, about 46 μL, about 48 μL, about 50 μL, or any range including and/or in between any two of these values.
Applying the first citric acid solution and/or second citric acid solution (when present) may include spraying and/or coating of the first and/or second citric acid solution on a surface of a dressing composition (such as by use of an air brush and/or an ultrasonic sprayer).
The dressing composition may be according to any embodiment of a wound dressing composition of the present technology with the exception that the dressing composition does not include the first layer and does not include the second layer—thus, the method provides the first layer and (when present and as discussed herein) the second layer. Thus, the dressing composition includes about 30 wt. % to about 95 wt. % of one or more of collagen and chitosan, where the collagen may have a weight-average molecular weight of about 5,000 to about 100,000 and chitosan, and about 30 wt. % to about 70 wt. % ORC (where the ORC may have a weight-average molecular weight of about 50,000 to about 1,000,000), where a weight ratio of the one or more of collagen and chitosan to ORC is about 70:30 to about 30:70. It is to be understood that any component, value, and range/sub-range discussed in regard to any embodiment the wound dressing composition (absent, of course the first layer and when present the second layer) may be included in the dressing composition. Purely by way of example, U.S. Pat. Publ. 2001/0045177 provides procedures and protocols for producing dressing compositions.
As discussed above, the citric acidic solution includes a solvent and about 50 mM to about 400 mM citric acid. Thus, the concentration of citric acid in the citric acid solution may be about 50 mM, about 60 mM, about 70 mM, about 80 mM, about 90 mM, about 100 mM, about 110 mM, about 120 mM, about 130 mM, about 140 mM, about 150 mM, about 160 mM, about 170 mM, about 180 mM, about 190 mM, about 200 mM, about 220 mM, about 240 mM, about 260 mM, about 280 mM, about 300 mM, about 320 mM, about 340 mM, about 360 mM, about 380 mM, about 400 mM, or any range including and/or in between any two of the these values.
The first solvent and second solvent may each independently include one or more of water, ethanol, acetone, acetic acid, or a mixture of any two or more thereof. For example, in any embodiment herein, the first solvent and the second solvent may each independently include water. The amount of water (based on the total weight of the respective citric acid solution not considering the weight of citric acid) may independently be for the first and second citric acid solution about 10 wt. %, about 15 wt. %, about 20 wt. %, about 25 wt. %, about 30 wt. %, about 35 wt. %, about 40 wt. %, about 45 wt. %, about 50 wt. %, about 55 wt. %, about 60 wt. %, about 65 wt. %, about 70 wt. %, about 75 wt. %, about 80 wt. %, about 85 wt. %, about 90 wt. %, about 95 wt. %, about 100 wt. %, or any range including or in between any two of these values.
In any embodiment disclosed herein, the first solvent and second solvent may each independently include from about 10% to about 95% ethanol. For example, in any embodiment herein, the first solvent and the second solvent may each independently include ethanol present in an amount of about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 95%, or any range including and/or in between any two of the preceding values. In any embodiment disclosed herein, the first solvent and second solvent may each independently include about 70% ethanol.
In any embodiment disclosed herein, the first solvent and second solvent may each independently include about 0.01 M to about 0.1 M acetic acid. Thus, in any embodiment herein, the concentration of acetic acid may be about 0.01 M, about 0.02 M, about 0.03 M, about 0.04 M, about 0.05 M, about 0.06 M, about 0.07 M, about 0.08 M, about 0.09 M, about 0.1 M, or any range including and/or in between any two of the preceding values. For example, in any embodiment herein, the first solvent and second solvent may each independently include 0.05 M acetic acid.
The first citric acid solution and second citric acid solution may further each independently include a dye marker, such as E124, fluorescein isothiocyanate (FITC), 4′,6-diamidino-2-phenylindole (DAPI), methylene blue, erythrosine B, ponceau S, alura red, SYBR green, alcian blue, brilliant blue G, calcein blue, cardio green, crystal violet, nile blue, india ink, brilliant blue, indigo carmine, sudan III, methyl green, oil red, pyronin Y, tattoo ink, purpurin, phloxine B, picric acid, carbon nanotubes, a fuchsin, resazurin, a trichrome, or a combination of any two or more thereof. In any embodiment herein, each dye marker may be independently included at a concentration of about 1 millimolar (mM) to about 100 mM. Thus, the concentration of each dye marker in each solution may independently be about 1 mM, about 2 mM, about 3 mM, about 4 mM, about 5 mM, about 6 mM, about 7 mM, about 8 mM, about 9 mM, about 10 mM, about 11 mM, about 12 mM, about 13 mM, about 14 mM, about 15 mM, about 16 mM, about 17 mM, about 18 mM, about 19 mM, about 20 mM, about 22 mM, about 24 mM, about 26 mM, about 28 mM, about 30 mM, about 32 mM, about 34 mM, about 36 mM, about 38 mM, about 40 mM, about 42 mM, about 44 mM, about 46 mM, about 48 mM, about 50 mM, about 55 mM, about 60 mM, about 65 mM, about 70 mM, about 75 mM, about 80 mM, about 85 mM, about 90 mM, about 95 mM, about 100 mM, or any range including and in between any two of these values.
The first citric acid solution and second citric acid solution may further each independently include an additive, such as acetic acid. In any embodiment herein, each additive may be independently included in the first citric acid solution at a concentration of about 0.01M to about 1 M. In any embodiment herein, each additive may be independently included in the second citric acid solution at a concentration of about 0.01M to about 1 M.
In any embodiment herein, the method may further include (subsequent to applying the first and (when present) second citric acid solution) a drying step to obtain the wound dressing composition. Such a drying step may be performed at a temperature of about 16° C. to about 40° C. In any embodiment disclosed herein, the method may include drying the citric acid while the dressing composition is under tension. Suitable temperatures in any embodiment herein may include, but are not limited to, about 16° C., about 17° C., about 18° C., about 19° C., about 20° C., about 21° C., about 22° C., about 23° C., about 24° C., about 25° C., about 26° C., about 27° C., about 28° C., about 29° C., about 30° C., about 31° C., about 32° C., about 33° C., about 34° C., about 35° C., about 36° C., about 37° C., about 38° C., about 39° C., about 40° C., or any range including and/or in between any two of the preceding values. For example, in any embodiment herein, the drying step may be performed at a temperature of about 20° C. to about 25° C.
In a related aspect, the present technology provides a wound dressing composition prepared according to the methods described herein in any embodiment.
In a further related aspect, the present disclosure provides kits that include a wound dressing composition of any embodiment described herein and instructions for use. The kit may optionally comprise components such as antiseptic wipes, ointment, adhesive tape, tweezers, scissors, etc. The instructions of any embodiment of a kit of the present technology may provide for a method of treating a wound in a subject.
The examples herein are provided to illustrate advantages of the present technology and to further assist a person of ordinary skill in the art with preparing or using the compositions and systems of the present technology. The examples should in no way be construed as limiting the scope of the present technology, as defined by the appended claims. The examples can include or incorporate any of the variations, aspects, or embodiments of the present technology described above. The variations, aspects, or embodiments described above may also further each include or incorporate the variations of any or all other variations, aspects or embodiments of the present technology.
The dressing of any embodiment described herein may be employed in therapy in which a tissue site is treated with reduced pressure. Treatment of tissue with reduced pressure may be commonly referred to as “negative-pressure therapy,” but is also known by other names, including “negative-pressure wound therapy,” “reduced-pressure therapy,” “vacuum therapy,” “vacuum-assisted closure,” and “topical negative-pressure,” for example. Negative-pressure therapy may provide a number of benefits, including migration of epithelial and subcutaneous tissues, improved blood flow, and/or micro-deformation of tissue at a wound site. Together, these benefits may increase development of granulation tissue and reduce healing times.
Generally, the system may be configured to provide negative-pressure to a tissue site in accordance with this specification. In any embodiment herein, the system may generally include a negative-pressure supply, and may include or be configured to be coupled to a distribution component. In general, a distribution component may refer to any complementary or ancillary component configured to be fluidly coupled to a negative-pressure supply in a fluid path between a negative-pressure supply and a tissue site.
In any embodiment herein, the dressing may be configured to distribute negative pressure. The dressing may comprise or be configured as a manifold. A “manifold” in this context generally includes any composition or structure providing a plurality of pathways configured to collect or distribute fluid across a tissue site under pressure. For example, a manifold may be configured to receive negative pressure from the negative-pressure source and to distribute negative pressure through multiple apertures (e.g., pores), which may have the effect of collecting fluid and drawing the fluid toward the negative-pressure source. Additionally or alternatively, the fluid path(s) may be reversed or a secondary fluid path may be provided to facilitate movement of fluid across a tissue site. Additionally or alternatively, the fluid pathways of a manifold may be interconnected to improve distribution or collection of fluids. Additionally or alternatively, a manifold may be a porous material having a plurality of interconnected cells or pores. For example, in any embodiment herein, open-cell foams such as reticulated foams may generally include pores, edges, and/or walls that may form interconnected fluid pathways (such as channels).
The fluid mechanics associated with using a negative-pressure source to reduce pressure in another component or location, such as within a sealed therapeutic environment, can be mathematically complex. However, the basic principles of fluid mechanics applicable to negative-pressure therapy are generally well-known to those skilled in the art. The process of reducing pressure may be described generally and illustratively herein as “delivering,” “distributing,” or “generating” negative pressure, for example.
In general, a fluid, such as wound fluid (for example, wound exudates and other fluids), flows toward lower pressure along a fluid path. Thus, the term “downstream” typically implies something in a fluid path relatively closer to a source of negative pressure or further away from a source of positive pressure. Conversely, the term “upstream” implies something relatively further away from a source of negative pressure or closer to a source of positive pressure. This orientation is generally presumed for purposes of describing various features and components herein. However, the fluid path may also be reversed in some applications (such as by substituting a positive-pressure source for a negative-pressure source) and this descriptive convention should not be construed as a limiting convention.
“Negative pressure” may generally refer to a pressure less than a local ambient pressure, such as the ambient pressure in a local environment external to a sealed therapeutic environment provided by the dressing. In many cases, the local ambient pressure may also be the atmospheric pressure proximate to or about a tissue site. Alternatively or additionally, the pressure may be less than a hydrostatic pressure associated with the tissue at the tissue site. While the amount and nature of negative pressure applied to a tissue site may vary according to therapeutic requirements, the pressure is generally a low vacuum, also commonly referred to as a rough vacuum, between −5 mm Hg (−667 Pa) and −500 mm Hg (−66.7 kPa), gauge pressure. Common therapeutic ranges are between −50 mm Hg (−6.7 kPa) and −300 mm Hg (−39.9 kPa), gauge pressure.
Additionally or alternatively, in any embodiment herein, a negative-pressure supply may be a reservoir of air at a negative pressure, or may be a manual or electrically-powered device that can reduce the pressure in a sealed volume, such as a vacuum pump, a suction pump, a wall suction port available at many healthcare facilities, or a micro-pump, for example. A negative-pressure supply may be housed within or used in conjunction with other components, such as sensors, processing units, alarm indicators, memory, databases, software, display devices, or user interfaces that further facilitate therapy. A negative-pressure source may be combined with a controller and other components into a therapy unit. A negative-pressure supply may also have one or more supply ports configured to facilitate coupling and de-coupling of the negative-pressure supply to one or more distribution components.
In any embodiment herein, components may be fluidly coupled to each other to provide a path for transferring fluids (i.e., liquid and/or gas) between the components. For example, components may be fluidly coupled through a fluid conductor, such as a tube. As used herein, the term “fluid conductor” may include a tube, pipe, hose, conduit, or other structure with one or more lumina or open passages adapted to convey a fluid between two ends thereof. Typically, a fluid conductor may be an elongated, cylindrical structure with some flexibility, but the geometry and rigidity may vary. Additionally or alternatively, in any embodiment herein, the negative-pressure source may be operatively coupled to the dressing via a dressing interface.
The present technology also provides a therapy method, where the therapy method includes positioning a wound dressing composition of any embodiment herein of the present technology adjacent to the tissue site. For example, in any embodiment herein, the wound dressing may be positioned proximate to the wound. The wound dressing may be used with any of a variety of wounds, such as those occurring from trauma, surgery, or disease. For example, the wound dressing may be placed within, over, on, or otherwise proximate to the tissue site.
In any embodiment herein, the process may include employing the wound dressing in the context of a negative-pressure therapy, where the negative-pressure therapy may include positioning the wound dressing proximate to the tissue site (e.g., a wound). For example, the various components of the dressing may be positioned with respect to the tissue site sequentially or, alternatively, may be positioned with respect to each other and then positioned with respect to the tissue site. The negative-pressure therapy may further comprise sealing the wound dressing to tissue surrounding the tissue site to form a sealed space. For example, the wound dressing may be positioned proximate to the tissue site and sealed to an attachment surface near the tissue site, for example, to undamaged epidermis peripheral to a tissue site.
The negative-pressure therapy method in any embodiment herein may further include fluidly coupling a negative-pressure source to the sealed space and operating the negative-pressure source to generate a negative pressure in the sealed space. For example, the negative-pressure source may be coupled to the dressing such that the negative-pressure source may be used to reduce the pressure in the sealed space. For example, negative pressure applied across the tissue site, for example, via the dressing may be effective to induce macrostrain and microstrain at the tissue site, as well as remove exudates and other fluids from the tissue site.
The present technology also provides a method of preventing formation of a biofilm or reducing biofilm at a tissue site, where the method includes applying a wound dressing composition of any embodiment herein of the present technology adjacent to the tissue site.
The following example illustrates how the compositions and methods of the present technology are surprisingly superior to wound dressing compositions provided by a common method of coating—dip coating.
In particular, dressing compositions Promogran® (“PN”; including a mixture of about 55 wt. % collagen and about 45 wt. % ORC) and Promogran Prisma® (“PA”; including a mixture of about 55 wt. % collagen, about 44 wt. % ORC, and about 1 wt. % of a combination of silver in ORC where the combination includes about 25% w/w ionically bound silver) were utilized in the following studies, where the compositional details of each are provided in Table 1 below.
The citric acid solutions employed in the study were
For the dip coating studies, the dry weight of each dressing was first determined, followed by dip coating the dressing composition for 2 minutes in one of various citric acid solutions or controls, and subsequently removing the dip-coated dressing composition from the solution/control and air drying at room temperature (about 23° C.) for about 24 hours. Once the dip-coated dressing composition was dry, it was re-weighed. The results of the dip coating studies are illustrated in Table 2.
For the spray coating studies, the dry weight of each dressing was first determined, followed by spray coating the one surface of the dressing (about 28 cm2 in surface area) with about 500 μL of one of the various citric acid solutions or controls, and subsequently allowing the spray-coated composition to air dry at room temperature (about 23° C.) for about 24 hours or oven dry at about 90° C. until dry (about 4-6 hours) under normal atmosphere. Once the spray-coated dressing composition was dry, it was re-weighed. The results of the spray coating studies are illustrated in Table 3 (for the air dried samples) and Table 4 (for the oven dried samples).
The results demonstrate that the dip-coated wound dressings increased substantially in weight after dip coating. The dressings also shrunk about 50% or more of the original size of the dressing after drying for about 24 hours at room temperature. Without wishing to be bound by theory, it is believed that the dip coating process with a citric acid solution is altering the characteristics of the wound dressing compositions after drying. The results of the spray coating experiments indicate that the weight of the wound dressings did not change much after drying, whether at room temperature or in an oven. Surprisingly, the wound dressing did not shrink as much in size after spray coating compared to the dip coating method. The dressing samples allowed to dry at room temperature only demonstrated about 10-15% shrinkage compared to the original size of the dressing. Dressing samples that were allowed to dry in the oven at about 90° C. demonstrated about 20-50% shrinkage along with curling of the wound dressing. These data suggest that that the spray coating technique was superior to dip coating regardless of whether the dressing is air-dried or oven dried, but that spray coating coupled with air drying is especially advantageous for incorporation of a layer of citric acid into a dressing composition.
The capability of wound dressing compositions disclosed herein in any embodiment (e.g., wound dressings prepared according to Example 1) may be assessed using a zone of inhibition assay in which the dressings are exposed to petri dishes having a lawn of bacteria already present. Such bacteria is allowed to grow and the antimicrobial activity of the respective wound dressing compositions may be assessed. In particular, the effect on biofilms of such wound dressing compositions may be evaluated to determine biofilm populations after 24 hours of exposure with each respective wound dressing composition. Overall reduction in biofilm populations (i.e., reduction in total viable count of the biofilm populations) following such exposure may be assessed and compared to untreated controls and comparative wound dressing compositions.
While certain embodiments have been illustrated and described, a person with ordinary skill in the art, after reading the foregoing specification, can effect changes, substitutions of equivalents and other types of alterations to the compositions, systems, and methods of the present technology. Each aspect and embodiment described above can also have included or incorporated therewith such variations or aspects as disclosed in regard to any or all of the other aspects and embodiments.
The present technology is also not to be limited in terms of the particular aspects and embodiments described herein, which are intended as single illustrations of individual aspects of the present technology. Many modifications and variations of this present technology can be made without departing from its spirit and scope, as will be apparent to those skilled in the art. Functionally equivalent methods within the scope of the present technology, in addition to those enumerated herein, will be apparent to those skilled in the art from the foregoing descriptions. Such modifications and variations are intended to fall within the scope of the appended claims. It is to be understood that this present technology is not limited to particular methods, reagents, compounds, compositions, labeled compounds or biological systems, which can, of course, vary. It is also to be understood that the terminology used herein is for the purpose of describing particular aspects only, and is not intended to be limiting. Thus, it is intended that the specification be considered as exemplary only with the breadth, scope and spirit of the present technology indicated only by the appended claims, definitions therein and any equivalents thereof.
The embodiments, illustratively described herein may suitably be practiced in the absence of any element or elements, limitation or limitations, not specifically disclosed herein. Thus, for example, the terms “comprising,” “including,” “containing,” etc. shall be read expansively and without limitation. Additionally, the terms and expressions employed herein have been used as terms of description and not of limitation, and there is no intention in the use of such terms and expressions of excluding any equivalents of the features shown and described or portions thereof, but it is recognized that various modifications are possible within the scope of the claimed technology. Additionally, the phrase “consisting essentially of” will be understood to include those elements specifically recited and those additional elements that do not materially affect the basic and novel characteristics of the claimed technology. The phrase “consisting of” excludes any element not specified.
In addition, where features or aspects of the disclosure are described in terms of Markush groups, those skilled in the art will recognize that the disclosure is also thereby described in terms of any individual member or subgroup of members of the Markush group. Each of the narrower species and subgeneric groupings falling within the generic disclosure also form part of the invention. This includes the generic description of the invention with a proviso or negative limitation removing any subject matter from the genus, regardless of whether or not the excised material is specifically recited herein.
As will be understood by one skilled in the art, for any and all purposes, particularly in terms of providing a written description, all ranges disclosed herein also encompass any and all possible subranges and combinations of subranges thereof. Any listed range can be easily recognized as sufficiently describing and enabling the same range being broken down into at least equal halves, thirds, quarters, fifths, tenths, etc. As a non-limiting example, each range discussed herein can be readily broken down into a lower third, middle third and upper third, etc. As will also be understood by one skilled in the art all language such as “up to,” “at least,” “down to,” and the like, include the number recited and refer to ranges which can be subsequently broken down into subranges as discussed above. As will also be understood by one skilled in the art all language such as “greater than,” “less than,” and the like, refer to ranges which can be subsequently broken down into subranges as discussed above. Finally, as will be understood by one skilled in the art, a range includes each individual member.
All publications, patent applications, issued patents, and other documents (for example, journals, articles and/or textbooks) referred to in this specification are herein incorporated by reference as if each individual publication, patent application, issued patent, or other document was specifically and individually indicated to be incorporated by reference in its entirety to the extent they are not inconsistent with the explicit teachings of this specification. Definitions that are contained in text incorporated by reference are excluded to the extent that they contradict definitions in this disclosure.
Other embodiments are set forth in the following claims, along with the full scope of equivalents to which such claims are entitled.
| Filing Document | Filing Date | Country | Kind |
|---|---|---|---|
| PCT/IB2020/056042 | 6/25/2020 | WO |
| Number | Date | Country | |
|---|---|---|---|
| 62868611 | Jun 2019 | US |
