WO2021240271A1 - Encapsulation of antimicrobial agents for advanced wound dressings - Google Patents

Abstract

The present disclosure relates generally to wound dressing compositions that include microcapsules, where the microcapsules include one or more agents are described herein. The wound dressing compositions includes a mixture of a collagen, an oxidized regenerated cellulose (ORC), and a microcapsule that includes at least one agent. Also disclosed herein are kits including the wound dressing compositions of the present technology, and instructions for use.

Description

ENCAPSULATION OF ANTIMICROBIAL AGENTS FOR ADVANCED WOUND DRESSINGS

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of priority to U S. Provisional Application No.

63/031,718, filed on May 29, 2020, which is incorporated herein by reference in its entirety.

TECHNICAL FIELD

[0002] The present technology relates generally to wound dressing compositions that include a mixture of a collagen, an oxidized regenerated cellulose (ORC), and a microcapsule that includes an agent. Such wound dressing compositions may be used to treat and/or prevent an infection in a wound upon application.

BACKGROUND

[0003] The following description of the background of the present technology is provided simply as an aid in understanding the present technology and is not admited to describe or constitute prior art to the present technology.

[0004] A wide variety of materials and devices, generally characterized as “dressings,” are generally known in the art for use in treating an injury or other disruption of tissue. Such wounds may be the result of trauma, surgery, or disease, and may affect skin or other tissues. In general, dressings may control bleeding, absorb wound exudate, ease pain, assist in debriding the wound, protect wound tissue from infection, or otherwise promote healing and protect the wound from further damage. Infections can retard wound healing and, if untreated, can result in tissue loss, systemic infections, septic shock, amputation, 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. Currently, there is an unmet need for wound dressing products that can, upon application to a wound, deliver antimicrobial agents over a period of time in order to prevent or reduce infections in a wound upon application, and over time.

SUMMARY

[0005] In an aspect, a wound dressing composition is provided that includes about 30 wt.% to about 95 wt.% of a collagen with a weight-average molecular weight of about 5,000 to about 100,000, about 30 wt.% to about 70 wt.% oxidized regenerated cellulose (ORC) with a weight-average molecular weight of about 50,000 to about 1,000,000, and a microcapsule that includes an agent encapsulated in a polymer.

[0006] In a related aspect, a method for treating a subject in need thereof is provided, where the method includes administering to the wound a wound dressing composition of any embodiment disclosed herein. [0007] In a further related aspect, a kit is provided that includes a wound dressing composition of any embodiment disclosed herein as well as instructions for use.

BRIEF DESCRIPTION OF THE DRAWINGS [0008] FIG. 1 provides a non-limiting image of an embodiment of a microcapsule of the present technology, where the particular amount of agent in the microcapsule was about 47 wt.%.

[0009] FIG. 2 provides a non-limiting image of an embodiment of a microcapsule of the present technology, synthesized using a co-axial nozzle technique with a 750 pm nozzle and using an alginate based solution, according to the working examples.

[0010] FIG. 3 provides a non-limiting image of an embodiment of a microcapsule of the present technology, synthesized using a co-axial nozzle technique with a 300 pm nozzle and using an alginate based solution, according to the working examples.

[0011] FIG. 4 provides a non-limiting image of an embodiment of a microcapsule of the present technology, synthesized using a co-axial nozzle technique with a 200 pm nozzle and using an alginate based solution, according to the working examples.

[0012] FIG. 5 provides a non-limiting image of an embodiment of a microcapsule of the present technology, synthesized using a co-axial nozzle technique with a 450 pm nozzle and using an alginate based solution, according to the working examples.

DETAILED DESCRIPTION

[0013] 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.

Definitions

[0014] 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. [0015] The following terms are used throughout as defined below.

[0016] 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.

[0017] 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 be understood to 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 disclosing “10 wt.%.”

[0018] 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.

[0019] The phrase “and/or” as used in the present disclosure will be understood to mean any one of the recited members individually or a combination of any two or more thereof - for example, “A, B, and/or C” would mean “A, B, C, A and B, A and C, or B and C.”

[0020] 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).

[0021] As used herein, the term “effective amount” refers to a quantity sufficient to achieve a desired therapeutic and/or prophylactic effect, e.g. , an amount which results in the decrease in a wound described herein or one or more signs or symptoms associated with a wound described herein. In the context of therapeutic or prophylactic applications, the amount of a composition administered to the subject will vary depending on the composition, the degree, type, and severity of the wound and on the characteristics of the individual. The compositions can also be administered in combination with one or more additional therapeutic compounds. In the methods described herein, the therapeutic compositions may be administered to a subject having one or more wounds.

[0022] The term “mammalian recombinant collagen” refers to collagen manufactured by culturing a non-human organism or mammalian or non-mammalian cells to express at least one exogenous gene encoding a collagen in the culturing system. The term “human recombinant collagen” refers to collagen manufactured by culturing a non-human organism or mammalian or non-mammalian cells to express at least one human gene encoding a collagen. The human recombinant collagen may be selected from the group consisting of collagen type I, type II, type III, type IV, type V, type VI, type VII, type VIII, type IX, type X, type XI, type XII, type XIII, type XIV, type XV, type XVI, type XVII, type XVIII, type XIX, type XX, type XXI, type XXII, type XXIII, type XXIV, type XXV, type XXVI, and type XXVII. The human recombinant collagen can be collagen of one type free of any other type, or can be a mixture of collagen types. Suitably, the human recombinant collagen comprises collagens selected from the group consisting of collagen type I, collagen type III, and mixtures thereof. The term "bovine recombinant collagen" refers to collagen manufactured by culturing a non-human organism or mammalian or non-mammalian cells to express at least one bovine gene encoding a collagen. The bovine recombinant collagen may be selected from the group consisting of collagen type I, type II, type III, and type IV. The bovine recombinant collagen can be collagen of one type free of any other type, or can be a mixture of collagen types. Suitably, the bovine recombinant collagen comprises collagens selected from the group consisting of collagen type I, collagen type III, and mixtures thereof.

[0023] 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.

[0024] 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.

[0025] “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, e.g., alleviated, reduced, cured, or placed in a state of remission. [0026] 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, or few time administrations for the treatment of an acute wound.

The Wound Dressing Composition of the Present Technology

[0027] A wound can become infected by microbes. An infected wound is a wound in which bacteria or other microorganisms have significantly colonized to the extent of causing a deterioration and delay in the healing of the wound. Thus, a reduction in bacterial colonization is vital in wound therapy.

[0028] The present disclosure is directed, in part, to wound dressing compositions that include a microcapsule that includes an agent (such as an antimicrobial agent), that may treat and/or prevent infections in a wound upon application, and overtime. Certain embodiments of the wound dressing compositions of the present technology advantageously exhibit improved stability by encapsulating the antimicrobial agents in a polymer. Certain embodiments of the microcapsules of the wound dressing compositions of the present technology are further be configured to provide a staged release of the wound modulating components.

[0029] Thus, in an aspect, a wound dressing composition is provided that includes about 30 wt.% to about 95 wt.% of a collagen with a weight-average molecular weight of about 5,000 to about 100,000, about 30 wt.% to about 70 wt.% oxidized regenerated cellulose (ORC) with a weight-average molecular weight of about 50,000 to about 1,000,000, and a microcapsule comprising an agent encapsulated in a polymer. In any embodiment herein, the wound dressing composition may be in the form of a foam or may be in the form of a casting film. FIG. 1 provides a non-limiting image of an embodiment of a microcapsule of the present technology, where the particular amount of agent in the microcapsule was about 47 wt.%.

[0030] In any embodiment disclosed herein, the wound dressing composition may include a plurality of microcapsules. In any embodiment disclosed herein, the microcapsules may have a diameter from about 0.02 pm to about 2,000 pm. Thus, the microcapsules may have a diameter from about 0.02 pm, about 0.03 pm, about 0.04 pm, about 0.05 pm, about 0.06 pm, about 0.07 pm, about 0.08 pm, about 0.09 pm, about 0.1 pm, about 0.11 pm, about 0.12 pm, about 0.13 pm, about 0.14 pm, about 0.15 pm, about 0.16 pm, about 0.17 pm, about 0.18 pm, about 0.19 pm, about 0.2 pm, about 0.22 pm, about 0.24 pm, about 0.26 pm, about 0.28 pm, about 0.3 pm, about 0.32 pm, about 0.34 pm, about 0.36 pm, about 0.38 pm, about 0.4 pm, about 0.42 pm, about 0.44 pm, about 0.46 pm, about 0.48 pm, about 0.5 pm, about 0.55 pm, about 0.6 pm, about 0.65 pm, about 0.7 pm, about 0.75 pm, about 0.8 pm, about 0.85 pm, about 0.9 pm, about 0.95 pm, about 1 pm, about 2 pm, about 3 pm, about 4 pm, about 5 pm, about 6 pm, about 7 pm, about 8 pm, about 9 pm, about 10 pm, about 11 pm, about 12 pm, about 14 pm, about 16 pm, about 18 pm, about 20 pm, about 22 pm, about 24 mih, about 26 mih, about 28 mhi, about 30 mih, about 32 mhi, about 34 mih, about 36 mhi, about 38 mhi, about 40 mhi, about 42 mhi, about 44 mhi, about 46 mhi, about 48 mhi, about 50 mhi, about 55 mhi, about 60 mhi, about 65 mhi, about 70 mhi, about 75 mih, about 80 mhi, about 85 mhi, about 90 mhi, about 95 pm, about 100 pm, about 150 mhi, about 200 mhi, about 250 mhi, about 300 mih, about 350 mhi, about 400 mhi, about 450 mih, about 500 ₍am, about 550 mhi, about 600 mhi, about 650 mm, about 700 mih, about 750 pm, about 800 mih, about 850 ₍am, about 900 ₍am, about 1,000 mih, about 1,100 mih, about 1,200 mpi, about 1,300 mih, about 1,400 mm, about 1,500 mhi, about 1,600 mm, about 1,700 mih, about 1,800 pm. about 1,900 mm, about 2,000 mm, or any range including and/or in between any two of these values. For example, the microcapsules may have a diameter from about 100 pm to about 2,000 pm. The diameter of the microcapsules of any embodiment disclosed herein may be a weight-average diameter.

[0031] In any embodiment disclosed herein, the wound dressing composition may include about 0.5 wt.% to about 50 wt.% microcapsules. Additionally or alternatively, in some embodiments, the wound dressing composition may include about 1 wt.% to about 10 wt.% microcapsules. Thus, the wound dressing composition may include microcapsules in an amount of 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 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 22 wt.%, about 24 wt.%, about 26 wt.%, about 28 wt.%, 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.%, or any range including and/or in between any two of these values.

[0032] In any embodiment disclosed herein, the microcapsule may include about 0.01 wt.% to about 40 wt.% of the agent based on the total weight of the microcapsule. Thus, the microcapsule may include the agent in an amount (based on the total weight of the microcapsule) of about 0.01 wt.%, about 0.05 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 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 22 wt.%, about 24 wt.%, about 26 wt.%, about 28 wt.%, about 30 wt.%, about 32 wt.%, about 34 wt.%, about 36 wt.%, about 38 wt.%, about 40 wt.%, or any range including and/or in between any two of these values.

[0033] In any embodiment disclosed herein, the microcapsules may be configured to release the agent(s) over a finite period of time, e g ., from about 2 days to about 14 days. Additionally or alternatively, in some embodiments, the microcapsules may be configured to release the agent(s) over a finite period of time that is greater than 10 days. Thus, the microcapsules may be configured to release the agent(s) over a period of time from about 2 days, about 3 days, about 4 days, about 5 days, about 6 days, about 7 days, about 8 days, about 9 days, about 10 days, about 11 days, about 12 days, about 13 days, about 14 days, or any range including and/or in between any two of these values.

[0034] In any embodiment disclosed herein, the polymer may include polyethylene glycol, polyvinyl alcohol, polyhydroxyethylmethacrylate, polylactic acid, chitosan, alginate, cellulose acetate, carrageenan, casein, gelatin, hyaluronic acid, silk, fibrin, dextrans, cyclodextrins, polycaprolactone, polyhydroxybutarate, polyhydroxyvalerate, poly(lactic-co-glycolic acid), or a combination of any two or more thereof.

[0035] In any embodiment disclosed herein, the agent may include an antimicrobial agent, a signaling protein, an antioxidant, or a combination of any two or more thereof.

[0036] In any embodiment disclosed herein, the antimicrobial agent may include acetic acid, benzoic acid, natamycin, nisin, citric acid, sorbic acid, polyhexamethylene biguanide, povidone iodine, propionic acid, honey, sulfites or any acid thereof, or a combination of any two or more thereof.

[0037] In any embodiment disclosed herein, the signaling protein may include a platelet-derived growth factor (PDGF), a hepatocyte growth factor (HGF), a transforming growth factor beta (TGFP). a fibroblast growth factor (FGF), an epidermal growth factor (EGF), keratinocyte growth factor (KGF), Caveolin-1 (Cavl), or a combination of any two or more thereof. Additionally or alternatively, in some embodiments, the signaling protein may include fibroblast growth factor 1 (FGF1), fibroblast growth factor 2 (FGF2), fibroblast growth factor 3 (FGF3), fibroblast growth factor 4 (FGF4), fibroblast growth factor 5 (FGF5), fibroblast growth factor 6 (FGF6), fibroblast growth factor 7/keratinocyte growth factor (FGF7/KGF), fibroblast growth factor 8 (FGF8), fibroblast growth factor 9 (FGF9), fibroblast growth factor 10/keratinocyte growth factor 2 (FGF10/KGF2), fibroblast growth factor 11 (FGF11), fibroblast growth factor 12 (FGF12), fibroblast growth factor 13 (FGF13), fibroblast growth factor 14 (FGF14), fibroblast growth factor 15 (FGF15), fibroblast growth factor 16 (FGF16), fibroblast growth factor 17 (FGF17), fibroblast growth factor 18 (FGF18), fibroblast growth factor 19 (FGF19), fibroblast growth factor 20 (FGF20), fibroblast growth factor 21 (FGF21), fibroblast growth factor 22 (FGF22), fibroblast growth factor 23 (FGF23), or a combination of any two or more thereof.

[0038] In any embodiment disclosed herein, the antioxidant may include anthocyanins, astaxanthin, bilirubin, canthaxanthin, capsaicin, citric acid, curcumin, coenzyme Q10, eugenol, flavanol, flavonolignans, flavanone, flavone, flavonol, iodide, isoflavone phytoestrogens, lutein, lycopene, manganese, melatonin, N-acetylcysteine, oxalic acid, phenolic acids, phytic acid, A-a-lipoic acid, stilbenoids, tocopherol, tocotrienol, vitamin A, vitamin C, vitamin E, xanthones, zeaxanthin, a- carotene, b-carotene, or a combination of any two or more thereof.

[0039] Additionally or alternatively, in some embodiments, the anthocyanins may include, but are not limited to, cyanidin, delphinidin, malvidin, pelargonidin, peonidin, petunidin, or a combination of any two or more thereof. [0040] Additionally or alternatively, in some embodiments, the flavanols may include, but are not limited to, catechin, epicatechin, theaflavin, thearubigins, gallocatechin, epigallocatechin, a gallate ester of any thereof, or a combination of any two or more thereof.

[0041] Additionally or alternatively, in some embodiments, the flavanones may include, but are not limited to, eriodictyol, hesperetin, naringenin, or a combination of any two or more thereof.

[0042] Additionally or alternatively, in some embodiments, the flavones may include, but are not limited to, apigenin, luteolin, tangeritin, or a combination of any two or more thereof.

[0043] Additionally or alternatively, in some embodiments, the flavonols may include, but are not limited to, isorhamnetin, kaempferol, myricetin, proanthocyanidins, quercetin, rutin, or a combination of any two or more thereof.

[0044] Additionally or alternatively, in some embodiments, the isoflavone phytoestrogens may include, but are not limited to, daidzein, genistein, glycitein, or a combination of any two or more thereof.

[0045] Additionally or alternatively, in some embodiments, the phenolic acids may 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, or a combination of any two or more thereof.

[0046] Additionally or alternatively, in some embodiments, the stillbenoids may include, but are not limited to, resveratrol, pterostilbene, or a combination thereof.

[0047] In any embodiment disclosed herein, the collagen may be recombinant or naturally occurring. Additionally or alternatively, in some embodiments, the collagen may be a mammalian collagen, an avian collagen, a reptilian collagen, an amphibian collagen, fish collagen, or a combination of any two or more thereof. Additionally or alternatively, in some embodiments, the collagen may be mammalian collagen. Additionally or alternatively, in some embodiments, the mammalian collagen may be a bovine collagen, an ovine collagen, a porcine collagen, a human collagen, or a combination of any two or more thereof. Additionally or alternatively, in some embodiments, the collagen may be a human collagen. Additionally or alternatively, in some embodiments, the human collagen may be human collagen type I, human collagen type III, or any combination thereof. Additionally or alternatively, in some embodiments, the collagen may be a bovine collagen. Additionally or alternatively, in some embodiments, the collagen may be bovine collagen type I, bovine collagen type II, bovine collagen type III, bovine collagen type IV, or a combination of any two or more thereof. Additionally or alternatively, in some embodiments, the collagen may be ovine collagen type I, ovine collagen type II, ovine collagen type III, ovine collagen type IV, or a combination of any two or more thereof. Additionally or alternatively, in some embodiments, the collagen may be porcine collagen type I, porcine collagen type II, porcine collagen type III, porcine collagen type IV, or a combination of any two or more thereof. Additionally or alternatively, in some embodiments, the collagen may be a combination of bovine collagen type I and bovine collagen type III. Additionally or alternatively, in some embodiments, the collagen may be type I human recombinant collagen, type III human recombinant collagen, type I bovine collagen, type II bovine collagen, or a combination of any two or more thereof.

[0048] In any embodiment disclosed herein, the collagen may be provided by any manner known in the art. Additionally or alternatively, in some embodiments, the collagen may be provided by a tissue sample or recombinantly manufactured. Additionally or alternatively, in some embodiments, mammalian recombinant collagen may be provided by any suitable method known in the art. Additionally or alternatively, in some embodiments, human recombinant collagen may be provided by any suitable method known in the art. For example, the step of providing human recombinant collagen may comprise following the protocol described in U.S. Pat. No. 5,962,648, the entire content of which is incorporated herein by reference. Further recombinant processes are set forth in U.S. Pat. No. 5,593,859 and W02004/078120, which are also incorporated herein by reference. Additionally or alternatively, in some embodiments, collagen will be recombinantly manufactured by culturing a cell which has been transfected with at least one gene encoding a polypeptide comprising collagen and genes encoding oxidized cellulose and subunits of the post-translational enzyme prolyl 4- hydroxylase, and purifying the resultant collagen monomer therefrom. Additionally or alternatively, in some embodiments, collagen will be recombinantly manufactured by a plant (e.g., CollPlant, CollPlant Holdings Ltd., Ness Ziona, Israel) such as tobacco, or in yeast. The human recombinant collagen solution may be subsequently subjected to polymerization or cross-linking conditions to produce an insoluble fibrous collagen.

[0049] In any embodiment disclosed herein, the collagen may be a type I collagen, a type II collagen, or a type III collagen. Additionally or alternatively, in some embodiments, the 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 any combination 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, the entire contents of which are incorporated by reference. [0050] As discussed above, the wound dressing composition includes about 30 wt.% to about 95 wt.% of a collagen. Thus, the collagen may be included in the wound dressing composition in an amount of about 1 wt.%, about 1.1 wt.%, about 1.2 wt.%, about 1.3 wt.%, about 1.4 wt.%, about 1.5 wt.%, about 1.6 wt.%, about 1.7 wt.%, about 1.8 wt.%, about 1.9 wt.%, about 2 wt.%, about 2.2 wt.%, about 2.4 wt.%, about 2.6 wt.%, about 2.8 wt.%, about 3 wt.%, about 3.2 wt.%, about 3.4 wt.%, about 3.6 wt.%, about 3.8 wt.%, about 4 wt.%, about 4.2 wt.%, about 4.4 wt.%, about 4.6 wt.%, about 4.8 wt.%, about 5 wt.%, about 5.2 wt.%, about 5.4 wt.%, about 5.6 wt.%, about 5.8 wt.%, about 6 wt.%, about 6.2 wt.%, about 6.4 wt.%, about 6.6 wt.%, about 6.8 wt.%, about 7 wt.%, about 7.2 wt.%, about 7.4 wt.%, about 7.6 wt.%, about 7.8 wt.%, about 8 wt.%, about 8.2 wt.%, about 8.4 wt.%, about 8.6 wt.%, about 8.8 wt.%, about 9 wt.%, about 9.2 wt.%, about 9.4 wt.%, about 9.6 wt.%, about 9.8 wt.%, about 10 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 22 wt.%, about 24 wt.%, about 26 wt.%, about 28 wt.%, 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 the preceding values.

[0051] In any embodiment disclosed herein, the collagen may have a weight-average molecular weight of about 5,000 to about 100,000. Additionally or alternatively, in some embodiments, 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 11,000, about 12,000, about 13,000, about 14,000, about 15,000, about 16,000, about 17,000, about 18,000, about 19,000, 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 42,000, about 44,000, about 46,000, about 48,000, about 50,000, about 52,000, about 54,000, about 56,000, about 58,000, about 60,000, about 62,000, about 64,000, about 66,000, about 68,000, about 70,000, about 72,000, about 74,000, about 76,000, about 78,000, about 80,000, about 82,000, about 84,000, about 86,000, about 88,000, about 90,000, about 92,000, about 94,000, about 96,000, about 98,000, about 100,000, or any range including and/or in between any two of the preceding values.

[0052] In any embodiment disclosed herein, the collagen of 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. Additionally or alternatively, in some embodiments, the ratio by weight of human collagen type I to human collagen type III is greater than about 50:50, or greater than about 70:30. Additionally or alternatively, in some embodiments, the collagen of the wound dressing composition may include a weight ratio of type I bovine collagen to type III bovine collagen of about 85:15.

[0053] In any embodiment disclosed herein, 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). Without wishing 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

[0054] In any embodiment disclosed herein, the wound dressing composition may include from about 30 wt.% to about 70 wt.% of ORC. Thus, ORC may be included in the wound dressing composition 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.

[0055] In any embodiment herein, the ORC may have a weight-average molecular weight of about 50,000 to about 1,000,000. Thus, the ORC may have a weight-average molecular weight of 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 110,000, about 120,000, about 130,000, about 140,000, about 150,000, about 160,000, about 170,000, about 180,000, about 190,000, about 200,000, about 210,000, about 220,000, about 230,000, about 240,000, about 250,000, about 260,000, about 270,000, about 280,000, about 290,000, about 300,000, about 310,000, about 320,000, about 330,000, about 340,000, about 350,000, about 360,000, about 370,000, about 380,000, about 390,000, about 400,000, about 410,000, about 420,000, about 430,000, about 440,000, about 450,000, about 460,000, about 470,000, about 480,000, about 490,000, about 500,000, about 510,000, about 520,000, about 530,000, about 540,000, about 550,000, about 560,000, about 570,000, about 580,000, about 590,000, about 600,000, about 610,000, about 620,000, about 630,000, about 640,000, about 650,000, about 660,000, about 670,000, about 680,000, about 690,000, about 700,000, about 710,000, about 720,000, about 730,000, about 740,000, about 750,000, about 760,000, about 770,000, about 780,000, about 790,000, about 800,000, about 810,000, about 820,000, about 830,000, about 840,000, about 850,000, about 860,000, about 870,000, about 880,000, about 890,000, about 900,000, about 910,000, about 920,000, about 930,000, about 940,000, about 950,000, about 960,000, about 970,000, about 980,000, about 990,000, about 1,000,000, or any range including and/or in between any two of the preceding values.

[0056] 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 pm to about 1,000 pm. In any embodiment herein, the ORC may include fiber lengths of about 5 pm, about 6 pm, about 7 pm, about 8 pm, about 9 pm, about 10 mih, about 11 mih, about 12 mhi, about 13 mih, about 14 mhi, about 15 mih, about 16 mhi, about 17 mhi, about 18 mhi, about 19 mhi, about 20 mhi, about 22 mhi, about 24 mhi, about 26 mhi, about 28 mhi, about 30 mhi, about 32 mhi, about 34 mhi, about 36 mih, about 38 mhi, about 40 mhi, about 42 mhi, about 44 pm, about 46 mih, about 48 mhi, about 50 mih, about 55 mhi, about 60 mhi, about 65 mih, about 70 pm, about 75 mih, about 80 mhi, about 85 mih, about 90 mhi, about 95 mhi, about 100 mhi, about 110 pm, about 120 pm, about 130 pm, about 140 mhi, about 150 mih, about 160 pm, about 170 mih, about 180 mih, about 190 mhi, about 200 mih, about 220 mhi, about 240 mpi, about 260 mpi, about 280 mhi, about 300 mih, about 320 mhi, about 340 mih, about 360 pm, about 380 mhi, about 400 mpi, about 420 mhi, about 440 mhi, about 460 mpi, about 480 mpi, about 500 mih, about 550 mpi, about 600 mhi, about 650 mhi, about 700 mhi, about 750 mhi, about 800 mhi, about 850 mhi, about 900 mpi, about 950 mhi, about 1,000 mhi, or any range including and/or in between any two of the preceding values. [0057] In any embodiment disclosed herein, the wound dressing composition may include a weight ratio of collagen to ORC of about 60:40 to about 40:60. Thus, the wound dressing composition may include a weight ratio of collagen to ORC of 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, or any range including and/or in between any two of these values. Additionally or alternatively, in some embodiments, in some embodiments, the weight ratio of the collagen to ORC may be about 55:45.

[0058] In any embodiment disclosed herein, the wound dressing composition may have a thickness of about 0.5 mm to about 5 mm. Thus, the wound dressing composition may have a thickness of about 0.5 cm, about 0.6 cm, about 0.7 cm, about 0.8 cm, about 0.9 cm, about 1 cm, about 1.1 cm, about 1.2 cm, about 1.3 cm, about 1.4 cm, about 1.5 cm, about 1.6 cm, about 1.7 cm, about 1.8 cm, about 1.9 cm, about 2 cm, about 2.1 cm, about 2.2 cm, about 2.3 cm, about 2.4 cm, about 2.5 cm, about 2.6 cm, about 2.7 cm, about 2.8 cm, about 2.9 cm, about 3 cm, about 3.1 cm, about 3.2 cm, about 3.3 cm, about 3.4 cm, about 3.5 cm, about 3.6 cm, about 3.7 cm, about 3.8 cm, about 3.9 cm, about 4 cm, about 4.1 cm, about 4.2 cm, about 4.3 cm, about 4.4 cm, about 4.5 cm, about 4.6 cm, about 4.7 cm, about 4.8 cm, about 4.9 cm, about 5 cm, or any range including and/or in between any two of these values.

[0059] In any embodiment disclosed herein, the wound dressing composition may have a solid content of about 0.1 wt.% to about 100 wt.%, such as about 0.1 wt.% to about 10 wt.%. Thus, the wound dressing composition may have a solid content of about 0.1 wt.%, about 0.11 wt.%, about 0.12 wt.%, about 0.13 wt.%, about 0.14 wt.%, about 0.15 wt.%, about 0.16 wt.%, about 0.17 wt.%, about 0.18 wt.%, about 0.19 wt.%, about 0.2 wt.%, about 0.22 wt.%, about 0.24 wt.%, about 0.26 wt.%, about 0.28 wt.%, about 0.3 wt.%, about 0.32 wt.%, about 0.34 wt.%, about 0.36 wt.%, about 0.38 wt.%, about 0.4 wt.%, about 0.42 wt.%, about 0.44 wt.%, about 0.46 wt.%, about 0.48 wt.%, about 0.5 wt.%, about 0.55 wt.%, about 0.6 wt.%, about 0.65 wt.%, about 0.7 wt.%, about 0.75 wt.%, about 0.8 wt.%, about 0.85 wt.%, about 0.9 wt.%, about 0.95 wt.%, about 1 wt.%, about 1.1 wt.%, about 1.2 wt.%, about 1.3 wt.%, about 1.4 wt.%, about 1.5 wt.%, about 1.6 wt.%, about 1.7 wt.%, about 1.8 wt.%, about 1.9 wt.%, about 2 wt.%, about 2.1 wt.%, about 2.2 wt.%, about 2.3 wt.%, about 2.4 wt.%, about 2.5 wt.%, about 2.6 wt.%, about 2.7 wt.%, about 2.8 wt.%, about 2.9 wt.%, about 3 wt.%, about

3.1 wt.%, about 3.2 wt.%, about 3.3 wt.%, about 3.4 wt.%, about 3.5 wt.%, about 3.6 wt.%, about 3.7 wt.%, about 3.8 wt.%, about 3.9 wt.%, about 4 wt.%, about 4.1 wt.%, about 4.2 wt.%, about 4.3 wt.%, about 4.4 wt.%, about 4.5 wt.%, about 4.6 wt.%, about 4.7 wt.%, about 4.8 wt.%, about 4.9 wt.%, about 5 wt.%, about 5.2 wt.%, about 5.4 wt.%, about 5.6 wt.%, about 5.8 wt.%, about 6 wt.%, about

6.2 wt.%, about 6.4 wt.%, about 6.6 wt.%, about 6.8 wt.%, about 7 wt.%, about 7.2 wt.%, about 7.4 wt.%, about 7.6 wt.%, about 7.8 wt.%, about 8 wt.%, about 8.2 wt.%, about 8.4 wt.%, about 8.6 wt.%, about 8.8 wt.%, about 9 wt.%, about 9.2 wt.%, about 9.4 wt.%, about 9.6 wt.%, about 9.8 wt.%, about 10 wt.%, about 20 wt.%, about 30 wt.%, about 40 wt.%, about 50 wt.%, about 60 wt.%, about 70 wt.%, about 80 wt.%, about 90 wt.%, about 95 wt.%, about 99 wt.%, about 100 wt.%, or any range including and/or in between any two of the preceding values.

[0060] In any embodiment disclosed herein, the wound dressing composition may include a silver compound. Additionally or alternatively, in some embodiments, the wound dressing composition may include about 0.1 wt.% to about 3 wt.% of the silver compound. Thus, the silver compound may be included in the wound dressing composition in an amount of about 0.1 wt.%, about 0.11 wt.%, about 0.12 wt.%, about 0.13 wt.%, about 0.14 wt.%, about 0.15 wt.%, about 0.16 wt.%, about 0.17 wt.%, about 0.18 wt.%, about 0.19 wt.%, about 0.2 wt.%, about 0.22 wt.%, about 0.24 wt.%, about 0.26 wt.%, about 0.28 wt.%, about 0.3 wt.%, about 0.32 wt.%, about 0.34 wt.%, about 0.36 wt.%, about 0.38 wt.%, about 0.4 wt.%, about 0.42 wt.%, about 0.44 wt.%, about 0.46 wt.%, about 0.48 wt.%, about 0.50 wt.%, about 0.52 wt.%, about 0.54 wt.%, about 0.56 wt.%, about 0.58 wt.%, about 0.6 wt.%, about 0.62 wt.%, about 0.64 wt.%, about 0.66 wt.%, about 0.68 wt.%, about 0.7 wt.%, about 0.72 wt.%, about 0.74 wt.%, about 0.76 wt.%, about 0.78 wt.%, about 0.8 wt.%, about 0.82 wt.%, about 0.84 wt.%, about 0.86 wt.%, about 0.88 wt.%, about 0.9 wt.%, about 0.92 wt.%, about 0.94 wt.%, about 0.96 wt.%, about 0.98 wt.%, about 1 wt.%, about 1.1 wt.%, about 1.15 wt.%, about

1.2 wt.%, about 1.25 wt.%, about 1.3 wt.%, about 1.35 wt.%, about 1.4 wt.%, about 1.45 wt.%, about

1.5 wt.%, about 1.55 wt.%, about 1.6 wt.%, about 1.65 wt.%, about 1.7 wt.%, about 1.75 wt.%, about

1.8 wt.%, about 1.85 wt.%, about 1.9 wt.%, about 1.95 wt.%, about 2 wt.%, about 2.05 wt.%, about 2.1 wt.%, about 2.15 wt.%, about 2.2 wt.%, about 2.25 wt.%, about 2.3 wt.%, about 2.35 wt.%, about

2.4 wt.%, about 2.45 wt.%, about 2.5 wt.%, about 2.55 wt.%, about 2.6 wt.%, about 2.65 wt.%, about

2.7 wt.%, about 2.75 wt.%, about 2.8 wt.%, about 2.85 wt.%, about 2.9 wt.%, about 2.95 wt.%, about

3 wt.%, or any range including and/or in between any two of the preceding values.

[0061] In any embodiment disclosed herein, the silver compound may be one or more pharmaceutically acceptable salts. Additionally or alternatively, in some embodiments, the one or more pharmaceutically acceptable silver salts may include, but are not limited to, silver oxide, 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 -aminobenzoate, silver / -aminosalicylatc. nanocrystalline silver, any pharmaceutically acceptable salt thereof, a silver oxysalt (e.g., Ag7NOn), or a combination of any two or more thereof

[0062] In any embodiment disclosed herein, the wound dressing composition of the present technology may be capable of preventing, reducing, inhibiting, or dismpting 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 comprise bacteria including, but not limited to, Pseudomonas aeruginosa, Staphylococcus aureus and Streptococcus mutans. The biofilm may also include fungi including but not limited to yeasts, such as Candida albicans. Additionally or alternatively, in some embodiments, the wound dressing composition of the present technology may be capable of preventing, reducing, inhibiting, or dismpting a biofilm in a wound by > about 10% to about 100%, after 24 hours in vitro exposure. Additionally or alternatively, in some embodiments, the wound dressing composition of the present technology may be capable of preventing, reducing, inhibiting, or dismpting 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.

[0063] Certain embodiments of the therapeutic efficacy of the wound dressing composition of the present technology are 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., etal., J. Wound Care, 19:220-226(2010), which is incorporated herein by reference).

[0064] In any embodiment disclosed herein, the wound dressing composition of the present disclosure may be sterile and packaged in a microorganism-impermeable container.

Methods of Making the Wound Dressing of the Present Technoloev

[0065] Based on the description of the wound dressing composition of the present technology, a person of ordinary skill in the art will understand how to generate compositions of the present technology.

[0066] For example, a 2 weight % slurry of collagen (55 % by weight) and ORC (45 % by weight) is prepared in an acetic acid solution (0.5 M). The suspension was mixed for at least 15 minutes to ensure uniform mixing. Then microcapsules containing desired cargo is added to this mixture. Briefly, the capsules are synthesized using coaxial nozzle technique using a 100 to 1000 micron diameter nozzle with oscillating frequency ranging from 100 to 2000 Hz at a inlet pressure ranging from 10 to 400 mbar. The mixture containing collagen, ORC, and encapsulated cago was briefly stirred for uniform mixing and subsequently degassed. The mixture was then poured into freeze drying trays.

The trays were maintained at -80 °C for at least 15 hours. Then the mixture was freeze dried at a reduced pressure (primary drying: shelf -10 °C, condenser -80 °C, 254 mtorr, 24 hours; secondary drying: shelf 30 °C, condenser -80 °C, 254 mtorr, 4 hours) to obtain the wound dressing. Negative-Pressure Therapy

[0067] The wound dressing composition of any embodiment described herein may be employed in therapy in which a wound 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.

[0068] Generally, the system may be configured to provide negative-pressure to a wound 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 wound.

[0069] In any embodiment herein, the wound dressing composition may be configured to distribute negative pressure. The wound dressing composition 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 wound 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 wound. 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).

[0070] 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.

[0071] 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.

[0072] “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 wound dressing composition. In many cases, the local ambient pressure may also be the atmospheric pressure proximate to or about a wound. Alternatively or additionally, the pressure may be less than a hydrostatic pressure associated with the tissue at the wound. While the amount and nature of negative pressure applied to a wound 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.

[0073] 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.

[0074] 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 wound dressing composition via a dressing interface. Treatment Methods of the Present Technology

[0075] In an aspect, methods for treating a wound in a subject in need thereof are provided, wherein the method includes administering to the wound a wound dressing composition of any embodiment disclosed herein. The wound may be an acute wound or a chronic wound. Exemplary chronic wounds include, but are not limited to, infectious wounds, venous ulcers, decubitis ulcers, and/or diabetic ulcers; exemplary acute wounds include, but are not limited to, surgical wounds, trauma wounds, bum wounds, and/or donor sites. In any embodiment disclosed herein, the wound dressing composition may protect the wound from infection. The infection may be a bacterial infection or a fungal infection. In any embodiment disclosed herein, the bacterial infection may be caused by gramnegative or gram-positive bacteria. In any embodiment disclosed herein, the subject may be a mammal, such as a human.

[0076] Examples of gram-positive bacteria include, but are not limited to, Actinomyces sp., Arcanobacterium sp., Bacillus sp., Bavariicoccus sp., Brachybacterium sp., Clostridium sp., Cnuibacter sp., Corynebacterium sp., Enterococcus sp., Desulfitobacterium sp., Fervidobacterium sp., Georgenia sp., Janibacter sp., Lactobacillales sp.,Microbispora sp., Nocardia sp., Pasteuria sp., Pilibacter sp., Propionibacterium sp., Rathayibacter sp., Rhodococcus sp., Roseburia sp., Rothia sp., Sarcina sp., Solibacillus sp., Sporosarcina sp., Staphylococcus sp., Streptococcus sp.,

Syntrophomonas sp., or Tepidibacter sp.

[0077] Examples of gram-negative bacteria include, but are not limited to, Acetobacter sp., Acidaminococcus sp. , Acinetobacter sp., Agrobacterium sp., Akkermansia sp., Anaerobiospirillum sp., Anaerolinea sp., Arcobacter sp., Armatimonas sp., Azotobacter sp., Bacteroides sp., Bacteroidetes sp., Bartonella sp., Bdellovibriosp., Brachyspira sp., Bradyrhizobium sp., Caldilinea sp., Cardiobacterium sp., Christensenella sp., Chthonomonas sp., Coxiella sp., Cyanobacteria sp., Cytophaga sp.,

Dehalo genimonas sp., De sulfur obacterium sp., Devosia sp., Dialister sp., Dictyoglomus sp., Dinoroseobacter sp., Enterobacter sp., Escherichia sp., Fimbriimonas sp., Flavobacterium sp., Francisella sp., Fusobacterium sp., Gluconacetobacter sp., Haemophilus sp., Helicobacter sp., Kingella sp., Klebsiella sp., Kluyvera sp., Kozakia sp., Legionella sp. Leptonema sp. Leptotrichia sp., Levilinea sp. Luteimonas sp. Megamonas sp.,Megasphaera sp.,Meiothermus sp., Methylobacterium sp., Moraxella sp., Morganella sp., Mycoplasma sp., Neisseria sp., Nitrosomonas sp., Pectinatus sp., Pedobacter sp., Pelosinus sp., Propionispora sp., Proteus sp., Pseudomonas sp., Pseudoxanthomonas sp., Rickettsia sp., Salinibacter sp., Salmonella sp., Samsonia sp., Serratia sp., Shigella sp.,

Shimwellia sp., Sphingomonas sp., Stenotrophomonas sp., Thorselliaceae sp., V ampirococcus sp., Verminephrobacter sp., Vibrio sp., Victivallis sp., Vitreoscilla sp., Wolbachia sp.

[0078] In any embodiment disclosed herein, the infection may be caused by Aspergillus sp., Aureobasidium sp., Candida sp., Cladosporium sp., Curvularia sp., Engodontium sp., Epicoccum sp., Gibberella sp., Hypocreales sp., Leptosphaerulina sp.,Malessezia sp., Penicillium sp., Rhodosporidium sp., Trichosporon sp., Trichtophyton sp., and Ulocladium sp. [0079] In any embodiment disclosed herein, the wound may include a biofilm, and the wound dressing composition of the present technology prevents, reduces, inhibits, or disrupts the biofdm. [0080] Additionally or alternatively, in some embodiments, the wound dressing composition may be administered directly to the wound. Any method known to those in the art for administering a wound dressing composition to an acute or a chronic wound disclosed herein may be employed. Suitable methods include in vitro or in vivo methods. In vivo methods typically include the administration of one or more wound dressing compositions to a subject in need thereof, suitably a human. When used in vivo for therapy, the one or more wound dressing compositions described herein are administered to the subject in effective amounts (i.e., amounts that have desired therapeutic effect). The dose and dosage regimen will depend upon the state of the wound of the subject, and the characteristics of the particular wound dressing composition used.

[0081] The effective amount may be determined during pre-clinical trials and clinical trials by methods familiar to physicians and clinicians. An effective amount of one or more wound dressing compositions useful in the methods may be administered to a subject in need thereof by any number of well-known methods for administering wound dressing compositions.

[0082] In any embodiment disclosed herein, the wound dressing compositions may be administered daily for 1 hour or more, for 2 hours or more, for 3 hours or more, for 4 hours or more, for 5 hours or more, for 6 hours or more, for 12 hours or more, or for 24 hours or more. In any embodiment disclosed herein, the wound dressing compositions may be administered one, two, three, four, or five times per day. In any embodiment disclosed herein, the wound dressing compositions may be administered daily for one, two, three, four or five weeks. For example, a wound dressing composition of any embodiment disclosed herein may be administered daily, where each daily administrating includes administering the wound dressing composition for 24 hours. In any embodiment disclosed herein, the wound dressing compositions may be administered daily for less than 6 weeks. In any embodiment disclosed herein, the wound dressing compositions may be administered daily for 6 weeks or more. In any embodiment disclosed herein, the wound dressing compositions may be administered daily for 12 weeks or more. In any embodiment disclosed herein, the wound dressing compositions may be administered every day, every other day, every third day, every fourth day, every fifth day, or every sixth day. In any embodiment disclosed herein, the wound dressing compositions may be administered weekly, bi-weekly, tri-weekly, or monthly. In any embodiment disclosed herein, the wound dressing compositions may be administered for a period of one, two, three, four, or five weeks. In any embodiment disclosed herein, the wound dressing compositions may be administered for six weeks or more. In any embodiment disclosed herein, the wound dressing compositions may be administered for twelve weeks or more. In any embodiment disclosed herein, the wound dressing compositions may be administered for a period of less than one year. In any embodiment disclosed herein, the wound dressing compositions may be administered for a period of more than one year. In any embodiment disclosed herein, the wound dressing compositions may be administered for an acute or chronic wound as appropriate.

[0083] In any embodiment herein, the process may further include employing the wound dressing in the context of a negative-pressure therapy, where the negative-pressure therapy may include positioning the wound dressing composition proximate to the wound. For example, the various components of the wound dressing composition may be positioned with respect to the wound sequentially or, alternatively, may be positioned with respect to each other and then positioned with respect to the wound. The negative-pressure therapy may further comprise sealing the wound dressing composition to tissue surrounding the wound to form a sealed space. For example, the wound dressing composition may be positioned proximate to the wound and sealed to an attachment surface near the wound, for example, to undamaged epidermis peripheral to a wound.

[0084] 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 wound dressing composition such that the negative -pressure source may be used to reduce the pressure in the sealed space. For example, negative pressure applied across the wound, for example, via the wound dressing composition may be effective to induce macrostrain and microstrain at the wound, as well as remove exudates and other fluids from the wound.

[0085] The present technology also provides a method of preventing formation of a biofilm or reducing biofilm at a wound, where the method includes applying a wound dressing composition of any embodiment herein of the present technology adjacent to the wound.

Kits Comprising the Wound Dressing of the Present Technology

[0086] 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 kits of the present technology may also include instructions for methods for treating a wound in a subject in need thereof. The kit may optionally comprise components such as antiseptic wipes, ointment, adhesive tape, tweezers, scissors, etc.

EXPERIMENTAL EXAMPLES

[0087] The present technology is further illustrated by the following examples, which should not be construed as limiting in any way.

Example 1 : Making the Microcapsules of the Present Technology

[0088] Exemplary microcapsules were synthesized using a co-axial nozzle technique. Specifically, antimicrobial additive containing solution is passed through the inner nozzle and alginate based solution is passed through the outer nozzle with nozzle diameter ranging between 100 to 1000 micron, voltage ranging from 1000 to 2500 V, vibrating frequency ranging from 100 to 2000 Hz, and inlet fluid flow pressure ranging from 100 to 400 mbarto obtain a final capsule with diameters ranging from 100 to 2000 microns by means of alginate crosslinking using Ca⁺² ions. Such exemplary microcapsules are illustrated in FIGs. 2-5, illustrating exemplary microcapsules formed via a 750 pm nozzle (FIG. 2), a 300 pm nozzle (FIG. 3), a 200 pm nozzle (FIG. 4), and a 450 pm nozzle (FIG. 5). Example 2: Disrupting Bio films in a Wound

[0089] A wound dressing composition of any embodiment described herein will be administered directly to a chronic wound. A first biofilm level will be determined using the colony drip flow reactor (C-DFR), described herein. A second biofilm level will be determined 72 hours after administering the wound dressing composition to the wound. It is anticipated that administration of the wound dressing composition of the present technology to a chronic wound will result in the prevention, reduction, inhibition, or disruption of biofilm levels in the wound.

[0090] These results will demonstrate that the wound dressing compositions of the present technology are useful for preventing, reducing, inhibiting, or disrupting biofilm levels in a wound in a subject in need thereof.

Example 3: Improved Collasen Synthesis Upon Application of the Wound Dressing Compositions of the Present Technology

[0091] A collagen synthesis assay with dermal fibroblasts is performed. This is a standard assay which shows the amount of collagen synthesized by fibroblasts after stimulation with the active agents in the wound dressing compositions of the present technology. Briefly, 8.4x 10⁴ human fibroblasts (per well) are plated into 24-well plates, and then incubated at 37° C, 5% CO2, in 10% FBS-DMEM. Once the cells are confluent (within 24 hours of plating), the 10% FBS-DMEM is removed, and the cells are washed 3^c with serum -free DMEM (SF-DMEM), before the test dressing samples of the present technology or a collagen/ORC alone dressing is added to the cells. Cells are then incubated for 72 hours after which time the media is collected and analyzed for the levels of the C-terminal propeptide of Type- 1 Collagen (Cl CP) present in the cell culture media. The level of Cl CP in the media, which is released by the fibroblasts as a by-product of collagen synthesis, is proportional to the level of collagen synthesis and so its level will be used to determine the level of collagen synthesis. It is anticipated that the wound dressing compositions of the present technology will exhibit increased collagen synthesis and/or improved wound healing of chronic or acute wounds compared to a standard collagen/ORC dressing.

[0092] Accordingly, the dressings of the present technology are useful in methods for treating a wound in a subject in need thereof, wherein the method comprises administering to the wound a dressing of any embodiment disclosed herein.

EQUIVALENTS

[0093] The present technology is not to be limited in terms of the particular embodiments described in this application, 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 and apparatuses 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 present technology. It is to be understood that this present technology is not limited to particular methods, reagents, compounds compositions, 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 embodiments only, and is not intended to be limiting.

[0094] 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.

[0095] 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,” “greater than,” “less than,” and the like, include the number recited and 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. Thus, for example, a group having 1-3 cells refers to groups having 1, 2, or 3 cells. Similarly, a group having 1-5 cells refers to groups having 1, 2, 3, 4, or 5 cells, and so forth.

[0096] All patents, patent applications, provisional applications, and publications referred to or cited herein are incorporated by reference in their entirety, including all figures and tables, to the extent they are not inconsistent with the explicit teachings of this specification.

Claims

1. A wound dressing composition comprising: about 20 wt.% to about 95 wt.% of a collagen with a weight-average molecular weight of about 5,000 to about 100,000; about 20 wt.% to about 70 wt.% oxidized regenerated cellulose (ORC) with a weight-average molecular weight of about 50,000 to about 1,000,000; and a microcapsule comprising an agent encapsulated in a polymer.

2. The wound dressing composition of claim 1, wherein a weight ratio of collagen to ORC in the wound dressing composition is about 60:40 to about 40:60.

3. The wound dressing composition of claim 1 or claim 2, wherein the wound dressing composition comprises a plurality of microcapsules.

4. The wound dressing composition of any one of claims 1-3, wherein the collagen is recombinant or naturally occurring.

5. The wound dressing composition of any one of claims 1-4, wherein the collagen is a mammalian collagen.

6. The wound dressing composition of claim 5, wherein the mammalian collagen comprises a bovine collagen, an ovine collagen, a porcine collagen, a human collagen, or a combination of any two or more thereof.

7. The wound dressing composition of any one of claims 1-6, wherein the collagen comprises type I human recombinant collagen, type III human recombinant collagen, type I bovine collagen, type III bovine collagen, type I ovine collagen, type III ovme collagen, type I porcine collagen, type III porcine collagen, or a combination of any two or more thereof.

8. The wound dressing composition of any one of claims 1-7, wherein the microcapsule has a diameter from about 0.02 pm to about 2,000 pm.

9. The wound dressing composition of any one of claims 1-8, wherein the wound dressing composition comprises about 0.5 wt.% to about 50 wt.% microcapsules.

10. The wound dressing composition of any one of claims 1-9, wherein the microcapsule comprises about 0.01 wt.% to about 40 wt.% of the agent based on the total weight of the microcapsule.

11. The wound dressing composition of any one of claims 1-10, wherein the polymer comprises polyethylene glycol, polyvinyl alcohol, polyhydroxyethylmethacrylate, polylactic acid, chitosan, alginate, cellulose acetate, carrageenan, casein, gelatin, hyaluronic acid, silk, fibrin, dextran, cyclodextrin, polycaprolactone, polyhydroxybutarate, polyhydroxyvalerate, poly(lactic-co-glycolic acid), or a combination of any two or more thereof.

12. The wound dressing composition of any one of claims 1-11, wherein the agent comprises an antimicrobial agent, a signaling protein, an antioxidant, or a combination of any two or more thereof.

13. The wound dressing composition of claim 12, wherein the antimicrobial agent comprises acetic acid, benzoic acid, natamycin, nisin, citric acid, sorbic acid, polyhexamethylene biguanide, povidone iodine, propionic acid, honey, sulfites or any acid thereof, a silver compound, or a combination of any two or more thereof.

14. The wound dressing composition of claim 12 or claim 13, wherein the signaling protein comprises a platelet-derived growth factor (PDGF), a hepatocyte growth factor (HGF), a transforming growth factor beta (TGFP). a fibroblast growth factor (FGF), an epidermal growth factor (EGF), keratinocyte growth factor (KGF), Caveolin-1 (Cavl), or a combination of any two or more thereof.

15. The wound dressing composition of any one of claims claim 12-14, wherein the signaling protein comprises fibroblast growth factor 1 (FGF1), fibroblast growth factor 2 (FGF2), fibroblast growth factor 3 (FGF3), fibroblast growth factor 4 (FGF4), fibroblast growth factor 5 (FGF5), fibroblast growth factor 6 (FGF6), fibroblast growth factor 7/keratinocyte growth factor (FGF7/KGF), fibroblast growth factor 8 (FGF8), fibroblast growth factor 9 (FGF9), fibroblast growth factor 10/keratinocyte growth factor 2 (FGF10/KGF2), fibroblast growth factor 11 (FGF11), fibroblast growth factor 12 (FGF12), fibroblast growth factor 13 (FGF13), fibroblast growth factor 14 (FGF14), fibroblast growth factor 15 (FGF15), fibroblast growth factor 16 (FGF 16), fibroblast growth factor 17 (FGF 17), fibroblast growth factor 18 (FGF 18), fibroblast growth factor 19 (FGF 19), fibroblast growth factor 20 (FGF20), fibroblast growth factor 21 (FGF21), fibroblast growth factor 22 (FGF22), fibroblast growth factor 23 (FGF23), or a combination of any two or more thereof.

16. The wound dressing composition of any one of claims 12-15, wherein the antioxidant comprises anthocyanin, astaxanthin, bilirubin, canthaxanthin, capsaicin, citric acid, curcumin, coenzyme Q10, eugenol, flavanol, flavonolignans, flavanone, flavone, flavonol, iodide, isoflavone phytoestrogens, lutein, lycopene, manganese, melatonin, N-acetylcysteine, oxalic acid, phenolic acids, phytic acid, /Z-a-lipoic acid, stilbenoids, tocopherol, tocotrienol, vitamin A, vitamin C, vitamin E, xanthones, zeaxanthin, a-carotene, b-carotene, or a combination of any two or more thereof.

17. The wound dressing composition of claim 16, wherein the anthocyanin comprises cyanidin, delphinidin, malvidin, pelargonidin, peonidin, petunidin, or a combination of any two or more thereof.

18. The wound dressing composition of claim 16 or claim 17, wherein the flavanol comprises catechin, epicatechin, theaflavin, thearubigins, gallocatechin, epigallocatechin, or any gallate ester thereof, or a combination of any two or more thereof.

19. The wound dressing composition of any one of claims 16-18, wherein the flavanone comprises eriodictyol, hesperetin, naringenin, or a combination of any two or more thereof.

20. The wound dressing composition of any one of claims 16-19, wherein the flavone comprises apigenin, luteolin, tangeritin, or a combination of any two or more thereof.

21. The wound dressing composition of any one of claims 16-20, wherein the flavonol comprises isorhamnetin, kaempferol, myricetin, proanthocyanidins, quercetin, rutin, or a combination of any two or more thereof.

22. The wound dressing composition of any one of claims 16-21, wherein the isoflavone phytoestrogen comprises daidzein, genistein, glycitein, or a combination of any two or more thereof.

23. The wound dressing composition of any one of claims 16-22, wherein the phenolic acid comprises chicoric acid, chlorogenic acid, cinnamic acid, ellagic acid, ellagitannins, gallic acid, gallotannins, rosmarinic acid, salicylic acid, or any ester thereof, or a combination of any two or more thereof.

24. The wound dressing composition of any one of claims 16-23, wherein the stilbenoid comprises resveratrol, pterostilbene, or a combination thereof.

25. The wound dressing composition of any one of claims 1-24, wherein the microcapsules are configured to release the agent over a period of time from about 2 days to about 14 days.

26. The wound dressing composition of any one of claims 1-25, wherein the wound dressing composition is in the form of a foam or in the form of a casting film.

27. A method for treating a wound in a subject in need thereof, the method comprising administering to the wound a wound dressing composition of any one of claims 1-26.

28. The method of claim 27, wherein the wound is an acute wound or a chronic wound.

29. The method of claim 27 or claim 28, wherein the wound dressing composition is administered directly to the wound.

30. The method of any one of claims 27-29, wherein the wound dressing composition prevents, reduces, inhibits, or disrupts biofilm formation in a wound.

31. The method of any one of claims 27-30, further comprising providing negative pressure therapy to the wound.

32. A kit comprising the wound dressing composition of any one of claims 1-26, and instructions for use.

33. The kit of claim 32, wherein the instructions for use comprise instructions for performing a method according to any one of claims 27 - 31.