PROCESS OF INCORPORATION OF FUNCTIONAU MOUECUUES WITHIN FREEZE-

DRIED DRESSING

CROSS-REFERENCE TO REUATED APPUICATIONS

[0001] This application claims the benefit of priority to U.S. Provisional Application No. 62/875,670, filed on July 18, 2019, which is incorporated herein by reference in its entirety.

TECHNICAU FIEUD

[0002] The present technology relates generally to wound dressing compositions including a mixture of a collagen and an oxidized cellulose and including a coating comprising functional molecules that is disposed adjacent to a wound-facing surface. Methods of manufacture and use as well as kits regarding the same are also provided.

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 admitted to describe or constitute prior art to the present technology.

[0004] The intricate process of wound healing involves the activation of numerous biological pathways that work in concert to regenerate a tissue microenvironment consisting of cells and extracellular matrix (ECM) with enzymes, cytokines, and growth factors. The wound healing process may be divided into the following sequential phases: hemostasis (clotting to stop bleeding), inflammatory response (removal of bacteria and tissue debris from the site of damage), proliferation (cell division to regenerate the tissue, angiogenesis, and matrix deposition), and remodeling (cellular apoptosis and matrix realignment in the newly generated tissue). These responses can be enhanced or accelerated by the delivery of functional molecules to the wound site, for example by coating a wound dressing with the desired functional molecules.

SUMMARY OF THE PRESENT TECHNOUOGY

[0005] In one aspect, the present disclosure provides a wound dressing composition comprising a freeze-dried layer with a wound-facing surface and an environmental-facing surface; wherein the freeze-dried layer comprises a first layer comprising a combination of collagen and an oxidized cellulose; and a coating forming a coated layer on the first layer, the coated layer comprising a formulation comprising an effective amount of at least one biologically active agent wherein the wound dressing composition does not visually appear to be wrinkled. Additionally or alternatively, in some embodiments, the formulation is applied to the first layer before freeze drying the first layer. [0006] Additionally or alternatively, in some embodiments, the coated layer has an average thickness of about 0.01 mih, about 0.03 mih, about 0.1 mih, about 0.3 mih, about 0.6 mih, about 1 mih, about 2 mih, about 5 mih, about 10 mih, about 15 mih, about 30 mih, about 50 mih, about 75 mm, about 100 mih, about 150 mih, about 200 mm, about 300 mih, about 400 mm, about 500 mih, about 600 mih, about 800 mm, or about 1,000 mih, or any range including and/or in between any two of these values. Additionally or alternatively, in some embodiments, the coated layer has an average thickness of about 0.1 mih to about 1 mih, about 0.3 mih to about 3 mih, about 1 mih to about 10 mih, about 3 mih to about 30 mih, about 10 mih to about 100 mih, about 30 mih to about 300 mih, or about 100 mih to about 1000 mih. Additionally or alternatively, in some embodiments, the wound dressing is resistant to structural changes compared to a control dressing that includes a collagen and an oxidized cellulose that has been freeze-dried and has been coated with the same formulation. In some embodiments, the control dressing is coated with the formulation via spray coating or coating with a brush.

[0007] In some embodiments, the oxidized cellulose may be oxidized regenerated cellulose (ORC).

[0008] Additionally or alternatively, in some embodiments, the formulation is applied at a temperature of about 40 °C, about 45 °C, about 50 °C, about 55 °C, about 60 °C, about 65 °C, about 70 °C, about 75 °C, about 80 °C, or any range including and/or in between any two of these values. Additionally or alternatively, in some embodiments, the formulation is applied at a temperature of about 40 °C to about 50 °C, about 45 °C to about 55 °C, about 50 °C to about 60 °C, about 55 °C to about 65 °C, about 60 °C to about 70 °C, about 65 °C to about 75 °C, about 70 °C to about 80 °C, or any range including and/or in between any two of these values.

[0009] Additionally or alternatively, in certain embodiments, the first layer is frozen at a temperature between about 0 °C to about -80 °C before application of the formula. In some embodiments, the first layer is frozen at a temperature of about 0 °C, about -5 °C, about -10 °C, about -15 °C, about -20 °C, about -30 °C, about -40 °C, about -50 °C, about -60 °C, about -70 °C, about -80 °C, or any range including and/or in between any two of these values. Additionally or alternatively, in some embodiments, the first layer is frozen at a temperature between about 0 °C to about -20 °C, about -10 °C to about -30 °C, about -20 °C to about -40 °C, about -30 °C to about -50 °C, about -40 °C to about -60 °C, about -50 °C to about -70 °C, about -60 °C to about -80 °C, or any range including and/or in between any two of these values.

[0010] Additionally or alternatively, in some embodiments, the first layer may include about 30 wt.% to about 70 wt.% oxidized cellulose. In some embodiments, the first layer may include oxidized cellulose at 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.%, or any range including and/or in between any two of the preceding values. Additionally or alternatively, in some embodiments, the first layer may include about 30 wt.% to about 40 wt.%, about 35 wt.% to about 45 wt.%, about 40 wt.% to about 50 wt.%, about 45 wt.% to about 55 wt.%, about 50 wt.% to about 60 wt.%, about 55 wt.% to about 65 wt.%, about 60 wt.% to about 70 wt.% oxidized cellulose, or any range including and/or in between any two of these values.

[0011] In certain embodiments, the oxidized cellulose may have a weight-average molecular weight of about 50,000 to about 1,000,000. Additionally or alternatively, in some embodiments, the oxidized cellulose may have a weight-average molecular weight of about 50,000, about 60,000, about 75,000, about 100,000, about 150,000, about 200,000, about 250,000, about 300,000, about 400,000, about 500,000, about 600,000, about 700,000, about 800,000, about 900,000, about 1,000,000, or any range including and/or in between any two of the preceding values. Additionally or alternatively, in some embodiments, the oxidized cellulose may have a weight-average molecular weight of about 50,000 to about 200,000, about 100,000 to about 250,000, about 200,000 to about 400,000, about 300,000 to about 500,000, about 400,000 to about 800,000, about 600,000 to about 1,000,000, or any range including and/or in between any two of these values.

[0012] In some embodiments, the collagen may be recombinant or naturally occurring. Additionally or alternatively, in some embodiments, the collagen may be a mammalian collagen. Additionally or alternatively, in some embodiments, the mammalian collagen may be a bovine collagen, a human collagen, or any combination thereof. Additionally or alternatively, in some embodiments, the collagen may comprise one or more of type I human recombinant collagen, type III human recombinant collagen, type I bovine collagen, type III bovine collagen, or any combination thereof.

[0013] In some embodiments, the first layer may include about 30 wt.% to about 95 wt.% collagen. Additionally or alternatively, in some embodiments, the first layer may include collagen at 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.% collagen, or any range including and/or in between any two of the preceding values. Additionally or alternatively, in some embodiments, the first layer may include about 30 wt.% to about 45 wt.%, about 40 wt.% to about 55 wt.%, about 50 wt.% to about 65 wt.%, about 60 wt.% to about 75 wt.%, about 70 wt.% to about 85 wt.%, about 80 wt.% to about 95 wt.% collagen, or any range including and/or in between any two of the preceding values.

[0014] In some embodiments, 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 7,500, about 10,000, about 15,000, about 20,000, about 25,000, about 30,000, about 40,000, about 50,000, about 60,000, about 70,000, about 80,000, about 90,000, about 100,000, or any range including and/or in between any two of the preceding values. Additionally or alternatively, in some embodiments, the collagen may have a weight- average molecular weight of about 5,000 to about 20,000, about 10,000 to about 25,000, about 20,000 to about 40,000, about 30,000 to about 50,000, about 40,000 to about 80,000, about 60,000 to about 100,000, or any range including and/or in between any two of these values. [0015] In some embodiments, the formulation may include a solvent and an effective amount of at least one agent selected from the group consisting of a growth factor, an antimicrobial agent, a peptide, a biopolymer, and any combination thereof. Additionally or alternatively, in some embodiments, the solvent may be an aqueous solvent or an organic solvent (e.g., water, citric acid, acetic acid etc.).

[0016] In some embodiments, the at least one agent may be included at about 0.01 wt.% to about 40 wt.%, or about 0.5 wt.% to about 5 wt.% in the formulation. Additionally or alternatively, in some embodiments, the at least one agent may be included at about 0.01 wt.%, about 0.025 wt.%, about 0.05 wt.%, about 0.1 wt.%, about 0.25 wt.%, about 0.5 wt.%, about 1 wt.%, about 5 wt.%, about 10 wt.%, about 15 wt.%, about 20 wt.%, about 25 wt.%, about 30 wt.%, about 35 wt.%, about 40 wt.% of the formulation, or any range including and/or in between any two of these values. Additionally or alternatively, in some embodiments, the at least one agent may be included about 0.01 wt.% to about 0.1 wt.%, about 0.05 wt.% to about 0.5 wt.%, about 0.1 wt.% to about 1 wt.%, about 0.5 wt.% to about 5 wt.%, about 1 wt.% to about 10 wt.%, about 5 wt.% to about 25 wt.%, about 10 wt.% to about 40 wt.% of the formulation, or any range including and/or in between any two of these values.

[0017] Additionally or alternatively, in some embodiments, the formulation may include one or more of a binder, a colorant, a deodorant agent, a fdler, a plasticizer, a viscosity modifier, an anti microbial, or any combination thereof. The binder may include one or more of hyaluronic acid, chitosan, heparin, alginate, cellulose, fibrin, gelatin, chondroitin sulfate, agarose, dextran, carrageenan, silk, polyethylene glycol), poly(vinyl alcohol), polycaprolactone, polyphophazene, polyglycolic acid, rosin, lactose, sucrose, tapioca starch, polyvinylpyrrolidone, or any combination thereof. In some embodiments, the colorant comprises one or more of direct red 80, bromophenol blue, toluidine blue, a food coloring, Allura Red AC, Erythrosine, Tartrazine 5, Anthocyanins, Curcuma, Annatto, or any combination thereof. Additionally or alternatively, in some embodiments, the deodorant agent comprises one or more of zinc ricinoleate, triethyl citrate, Saccharomyces ferment, Arctium lappa root extract, Humulus lupulus extract, Thymus vulgaris, or any combination thereof. Additionally or alternatively, in certain embodiments, the filler comprises one or more of amorphous silica, mica, kaolin, titanium dioxide, or any combination thereof.

[0018] Additionally or alternatively, in some embodiments, the plasticizer may include one or more of an acetylated monoglyceride, an alkyl citrate, methyl ricinoleate, glycerol, or any combination thereof. In some embodiments, the alkyl citrate may be one or more of triethyl citrate, acetyl triethyl citrate, tributyl citrate, acetyl tributyl citrate, trioctyl citrate, acetyl trioctyl citrate, trihexyl citrate, acetyl trihexyl citrate, butyryl trihexyl citrate, trimethyl citrate, or any combination thereof.

[0019] In some embodiments, the viscosity modifier may include one or more of sodium polyacrylate, acrylates/alkyl methacrylate block copolymers, polyacrylic acid, cellulose gum, hydroxyethylcellulose, stearyl alcohol, carbomer, or any combination thereof.

[0020] In some embodiments, the growth factor may include one or more of epidermal growth factor (EGF), hepatocyte growth factor (HGF), fibroblast growth factors (FGFs), platelet-derived growth factor (PDGF), or any combination thereof. Additionally or alternatively, in some embodiments, the fibroblast growth factors comprise one or more of 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 any combination thereof.

[0021] In some embodiments, the antimicrobial agent may include one or more of polyhexamethylene biguanide, acetic acid, benzoic acid, povidone iodine, natamycin, nisin, citric acid, sorbic acid, propionic acid, honey, sulfites, or any combination thereof.

[0022] In some embodiments, the peptide may include one or more of defensins, histatins, cathebcidin LL-37, or any combination thereof.

[0023] In some embodiments, the biopolymer may include one or more of hyaluronic acid, fibrinogen, a chitosan, a polysaccharide, a gelatin, a collagen, an oxidized cellulose, an oxidized regenerated cellulose (ORC), or any combination thereof.

[0024] In another aspect, the present disclosure provides a method for making a wound dressing composition of any embodiment disclosed herein, the method comprising, heating a formulation to about 40 °C to about 80 °C, depositing the formulation onto a layer comprising a collagen and an oxidized cellulose to form a coated layer, and freezing the coated layer, wherein the coated layer has an average thickness of about 0.1 pm to about 1,000 pm. Additionally or alternatively, in some embodiments, the method further comprises drying the coated layer.

[0025] Additionally or alternatively, in some embodiments, the coated layer has an average thickness of about 0.1 pm, about 0.3 pm, about 0.6 pm, about 1 pm, about 2 pm, about 5 pm, about 10 pm, about 15 pm, about 30 pm, about 50 pm, about 75 pm, about 100 pm, about 150 pm, about 200 pm, about 300 pm, about 400 pm, about 500 pm, about 600 pm, about 800 pm, or about 1,000 pm, or any range including and/or in between any two of these values. Additionally or alternatively, in some embodiments, the coated layer has an average thickness of about 0.1 pm to about 1 pm, about 0.3 pm to about 3 pm, about 1 pm to about 10 pm, about 3 pm to about 30 pm, about 10 pm to about 100 pm, about 30 pm to about 300 pm, or about 100 pm to about 1000 pm.

[0026] In some embodiments, the method comprises heating the formulation to about 40 °C, about 45 °C, about 50 °C, about 55 °C, about 60 °C, about 65 °C, about 70 °C, about 75 °C, about 80 °C, or any range including and/or in between any two of these values. Additionally or alternatively, in some embodiments, the method comprises heating the formulation to about 40 °C to about 50 °C, about 45 °C to about 55 °C, about 50 °C to about 60 °C, about 55 °C to about 65 °C, about 60 °C to about 70 °C, about 65 °C to about 75 °C, about 70 °C to about 80 °C, or any range including and/or in between any two of these values.

[0027] In some embodiments, the formulation is heated for a period between about 0.5 min to about 240 min. Additionally or alternatively, in some embodiments, the formulation is heated for a period between about 0.5 min, about 1 min, about 2.5 min, about 5 min, about 7.5 min, about 10 min, about 15 min, about 30 min, about 45 min, about 60 min, about 90 min, about 120 min, about 150 min, about 180 min, about 210 min, about 240 min, or any range including and/or in between any two of these values. Additionally or alternatively, in some embodiments, the formulation is heated for a period between about 0.5 min to about 5 min, about 2 min to about 20 min, about 5 min to about 50 min, about 20 min to about 90 min, about 45 min to about 180 min, about 60 min to about 240 min, or any range including and/or in between any two of these values.

[0028] In some embodiments, the first layer may be frozen to form a frozen layer prior to deposition of the formulation, wherein the frozen layer optionally comprises a first surface.

[0029] In some embodiments, the first surface of the frozen layer may be melted prior to deposition of the formulation.

[0030] In some embodiments, the formulation may be deposited by a spray process or a thin layer application. Additionally or alternatively, in some embodiments, the thin layer application may be carried out by means of a brush.

[0031] In some embodiments, the coated layer is frozen at a temperature between about 0 °C to about -80 °C. Additionally or alternatively, in some embodiments, the coated layer is frozen at a temperature of about 0 °C, about -5 °C, about -10 °C, about -15 °C, about -20 °C, about -30 °C, about -40 °C, about -50 °C, about -60 °C, about -70 °C, about -80 °C, or any range including and/or in between any two of these values. Additionally or alternatively, in some embodiments, the coated layer is frozen at a temperature between about 0 °C to about -20 °C, about -10 °C to about -30 °C, about -20 °C to about -40 °C, about -30 °C to about -50 °C, about -40 °C to about -60 °C, about -50 °C to about -70 °C, about -60 °C to about -80 °C, or any range including and/or in between any two of these values.

[0032] In some embodiments, the coated layer may be frozen and/or freeze-dried for a period between about 0.1 min to about 10,000 min. Additionally or alternatively, in some embodiments, the coated layer may be frozen and/or freeze-dried for a period between about 0.1 min, about 0.5 min, about 1 min, about 5 min, about 15 min, about 30 min, about 60 min, about 90 min, about 120 min, about 200 min, about 500 min, about 750 min, about 1000 min, about 2000 min, about 4000 min, about 6000 min, about 8000 min, about 10,000 min, or any range including and/or in between any two of these values. Additionally or alternatively, in some embodiments, the coated layer may be frozen and/or freeze-dried for a period between about 0.1 min to about 1 min, about 0.5 min to about 5 min, about 2 min to about 20 min, about 5 min to about 50 min, about 20 min to about 200 min, about 50 min to about 500 min, about 200 min to about 2000 min, about 500 min to about 5000 min, about 1000 min to about 10000 min, or any range including and/or in between any two of these values. Additionally or alternatively, in some embodiments, the coated layer may be frozen and/or freeze-dried for at least 30 minutes.

[0033] In some embodiments, the present disclosure provides for a wound dressing composition generated by any method disclosed herein.

[0034] In one aspect, the present disclosure provides a method for treating a wound in a subject in need thereof, the method including administering to the wound a wound dressing composition of any embodiment disclosed herein. Additionally or alternatively in some embodiments, the wound dressing composition induces, enhances, or promotes healing in the subject. Additionally or alternatively, in some embodiments, the subject is human.

[0035] In one aspect, the present disclosure provides a method for preventing or reducing the risk of wound infection in a subject in need thereof, the method including administering to the wound a wound dressing composition of any embodiment disclosed herein. Additionally or alternatively in some embodiments, the wound dressing composition induces, enhances, or promotes healing in the subject. Additionally or alternatively, in some embodiments, the subject is human. Additionally or alternatively, in some embodiments, the method further comprises performing negative pressure wound therapy to the wound prior to, concurrently with or after applying the wound dressing.

[0036] Also provided herein are kits comprising the wound dressing compositions of any embodiment described herein and instructions for use. Additionally or alternatively, in some embodiments, the kit, further comprises negative pressure wound therapy device.

[0037] In one aspect, the present disclosure provides an apparatus comprising a layer with a wound facing surface and an environmental-facing surface, wherein the freeze-dried layer comprises a first layer comprising a combination of collagen and an oxidized cellulose; and a coating forming a coated layer on the first layer, the coated layer comprising a formulation comprising an effective amount of at least one biologically active agent wherein the wound dressing composition does not visually appear to be wrinkled. Additionally or alternatively, in some embodiments, the formulation is applied to the first layer before freeze drying the first layer. Additionally or alternatively, in some embodiments, the coated layer has an average thickness of about 0.1 pm to about 1,000 pm. Additionally or alternatively, in some embodiments, the formulation is applied at a temperature of about 40 °C to about 80 °C. Additionally or alternatively, in some embodiments, the apparatus is resistant to structural changes compared to a control apparatus that includes a collagen and an oxidized cellulose that has been freeze- dried before coating with the same formulation.

[0038] In one aspect, the present disclosure provides a device for the treatment of wound comprising a layer a layer with a wound-facing surface and an environmental-facing surface, wherein the freeze-dried layer comprises a first layer comprising a combination of collagen and an oxidized cellulose; and a coating forming a coated layer on the first layer, the coated layer comprising a formulation comprising an effective amount of at least one biologically active agent wherein the wound dressing composition does not visually appear to be wrinkled. Additionally or alternatively, in some embodiments, the formulation is applied to the first layer before freeze drying the first layer. Additionally or alternatively, in some embodiments, the coated layer has an average thickness of about 0.1 pm to about 1,000 pm. Additionally or alternatively, in some embodiments, the formulation is applied at a temperature of about 40 °C to about 80 °C. Additionally or alternatively, in some embodiments, the device is resistant to structural changes compared to a device dressing that includes a collagen and an oxidized cellulose that has been freeze-dried before coating with the same formulation.

[0039] In one aspect, the present disclosure provides a method of the treatment of a wound in a subject in need thereof, the method comprising administering to the subject the apparatus disclosed herein or the device for the treatment of wound of any embodiment. Additionally or alternatively, in some embodiments, the method further comprises performing negative pressure wound therapy to the wound prior to, concurrently with or after applying the wound dressing.

[0040] In one aspect, the present disclosure provides a method for preventing or reducing the risk of wound infection in a subject in need thereof, the method comprising administering to the subject the apparatus disclosed herein or the device for the treatment of wound of any embodiment.

BRIEF DESCRIPTION OF THE DRAWINGS

[0041] FIG. l shows a flow diagram of exemplary methods to manufacture the wound dressing compositions of the present technology.

[0042] FIG. 2 shows non-limiting representations of specific embodiments of a wound dressing composition of the present technology.

[0043] FIG. 3 shows decreased wrinkle formation in the wound dressing composition of the present technology (right panel), compared to a wound dressing composition that has been initially freeze-dried prior to applying the citric acid coating (left panel).

DETAILED DESCRIPTION

[0044] 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.

[0045] The present disclosure is directed to wound dressing compositions which have been coated with a formulation to incorporate functional molecules onto the surface of the wound dressing compositions, and methods for making the same. In some instances, the wound dressing compositions of the present technology do not comprise any wrinkles after coating with the formulation, which can be beneficial for ease of use, product packaging and handling, and can decrease the rate of degradation of the wound dressing upon application to a wound (see Kharkar, P.M. et al. Chemical Society Reviews 42:7335-7372 (2013)). [0046] In one aspect, the present disclosure provides a wound dressing composition comprising a freeze-dried layer with a wound-facing surface and an environmental-facing surface; wherein the freeze-dried layer comprises a first layer comprising a combination of collagen and an oxidized cellulose; and a coating forming a coated layer on the first layer, the coated layer comprising a formulation comprising an effective amount of at least one biologically active agent wherein the wound dressing composition does not visually appear to be wrinkled. Additionally or alternatively, in some embodiments, the formulation the formulation is applied at a temperature of about 40 °C to about 80 °C. Additionally or alternatively, in some embodiments, the formulation is applied to the first layer before freeze drying the first layer. Additionally or alternatively, in some embodiments, the formulation the coated layer has an average thickness of about 0.1 pm to about 1,000 pm. Additionally or alternatively, in some embodiments, the formulation the wound dressing is resistant to structural changes compared to a control dressing that includes a collagen and an oxidized cellulose that has been freeze-dried and has been coated with the same formulation. In some embodiments, the control dressing is coated with the formulation via spray coating or continuous coating with a brush.

[0047] In another aspect, the present disclosure is directed to methods for making a wound dressing composition, the method comprising, heating a formulation to about 40 °C to about 80 °C, depositing the formulation onto a layer comprising a collagen and an oxidized cellulose to form a coated layer, and freezing the coated layer; wherein the coated layer has an average thickness of about 0.1 pm to about 1,000 pm.

[0048] FIG. 1 provides a representative, non-limiting flow diagram of an exemplary method for making the wound dressing compositions of the present technology. FIG. 2 provides representative, non-limiting illustrations of embodiments of the wound dressing compositions of the present technology. FIG. 3 demonstrates that the wound dressing compositions of the present technology feature decreased wrinkle formation compared to the wound dressing compositions which undergo freeze-drying prior to the application of the coating.

Definitions

[0049] 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.

[0050] The following terms are used throughout as defined below.

[0051] 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.

[0052] 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.

[0053] As used herein, the“administration” of a wound dressing composition to a subject includes any route of introducing or delivering to a subject a 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.

[0054] As used herein, the terms“contain”,“contains”, or“containing” in the context of describing the elements (especially in the context of the following claims) are to be construed as comprising or including the elements being described herein.

[0055] As used herein, the term“effective amount” refers to a quantity sufficient to achieve a desired therapeutic effect 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 wound dressing 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.

[0056] 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.

[0057] As understood by one of ordinary skill in the art,“molecular weight” (also known as “relative molar mass”) is a dimensionless quantity that can be 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.

[0058] As used herein, the term“solid content” refers to the density of a material and/or fdm of the wound dressing composition of the present technology, which is its mass per unit volume.

[0059] “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.

[0060] As used herein, the term“% w/v” refers to the percent of weight of the solution in the total volume of the solution, i.e., the number of grams of solute in 100 mL of solution.

[0061] 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 several administrations for the treatment of an acute wound.

The Wound Dressing Compositions of the Present Technology

[0062] The present disclosure provides a wound dressing composition comprising a layer with a wound-facing surface and an environmental-facing surface, wherein the first layer comprises a collagen and an oxidized cellulose, and wherein the first layer is coated with a formulation on the wound-facing surface to form a coated layer on the first layer; wherein the coated layer has an average thickness of about 0.1 pm to about 1,000 pm, the formulation is applied at a temperature of about 40 °C to about 80 °C, and the wound dressing is resistant to structural changes compared to a control dressing that includes a collagen and an oxidized cellulose, and has been freeze-dried and then coated with the same formulation. In any embodiment disclosed herein, the method further comprises drying the coated layer.

[0063] Additionally or alternatively, in some embodiments, the coated layer has an average thickness of about 0.1 pm, about 0.3 pm, about 0.6 pm, about 1 pm, about 2 pm, about 5 pm, about 10 pm, about 15 pm, about 30 pm, about 50 pm, about 75 pm, about 100 pm, about 150 pm, about 200 pm, about 300 pm, about 400 pm, about 500 pm, about 600 pm, about 800 pm, or about 1,000 pm, or any range including and/or in between any two of these values. Additionally or alternatively, in some embodiments, the coated layer has an average thickness of about 0.1 pm to about 1 pm, about 0.3 pm to about 3 pm, about 1 pm to about 10 pm, about 3 pm to about 30 pm, about 10 pm to about 100 pm, about 30 pm to about 300 pm, or about 100 pm to about 1000 pm.

[0064] In any embodiment disclosed herein, the formulation is applied at a temperature of about 40 °C to about 80 °C. In any embodiment disclosed herein, the formulation is applied at a temperature of about 40 °C, about 45 °C, about 50 °C, about 55 °C, about 60 °C, about 65 °C, about 70 °C, about 75 °C, about 80 °C, or any range including and/or in between any two of these values. Additionally or alternatively, in some embodiments, the formulation is applied at a temperature of about 40 °C to about 50 °C, about 45 °C to about 55 °C, about 50 °C to about 60 °C, about 55 °C to about 65 °C, about 60 °C to about 70 °C, about 65 °C to about 75 °C, about 70 °C to about 80 °C, or any range including and/or in between any two of these values. [0065] 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, or any combination thereof. Additionally or alternatively, in some embodiments, the collagen may be mammalian collagen. 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 any combination thereof. Additionally or alternatively, in some embodiments, the collagen may be bovine collagen type I and bovine collagen type III.

[0066] 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.

[0067] 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. [0068] In any embodiment disclosed herein, the first layer may include about 1 wt.% to about 95 wt.% collagen. Additionally or alternatively, in some embodiments, the first layer may include about 1 wt.%, about 2 wt.%, about 5 wt.%, 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.%, or any range including and/or in between any two of the preceding values, of collagen. Additionally or alternatively, in some embodiments, the first layer may include about 1 wt.% to about 10 wt.%, about 5 wt.% to about 20 wt.%, 10 wt.% to about 40 wt.%, about 30 wt.% to about 45 wt.%, about 40 wt.% to about 55 wt.%, about 50 wt.% to about 65 wt.%, about 60 wt.% to about 75 wt.%, about 70 wt.% to about 85 wt.%, about 80 wt.% to about 95 wt.% collagen, or any range including and/or in between any two of the preceding values.

[0069] In some embodiments, 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 7,500, about 10,000, about 15,000, about 20,000, about 25,000, about 30,000, about 40,000, about 50,000, about 60,000, about 70,000, about 80,000, about 90,000, about 100,000, or any range including and/or in between any two of the preceding values. Additionally or alternatively, in some embodiments, the collagen may have a weight- average molecular weight of about 5,000 to about 20,000, about 10,000 to about 25,000, about 20,000 to about 40,000, about 30,000 to about 50,000, about 40,000 to about 80,000, about 60,000 to about 100,000, or any range including and/or in between any two of these values.

[0070] In any embodiment disclosed herein, the oxidized cellulose comprises cellulose or a modified form of cellulose such as oxidized regenerated cellulose (ORC). Additionally or alternatively in some embodiments, the oxidized cellulose comprises ORC. In any embodiment disclosed herein, 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 need 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.

[0071] In any embodiment disclosed herein, the first layer may include about 30 wt.% to about 70 wt.% oxidized cellulose. In any embodiment disclosed herein, the first layer may include 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.%, or any range including and/or in between any two of the preceding values, of oxidized cellulose. In any embodiment disclosed herein, the first layer may include about 30 wt.% to about 40 wt.%, about 35 wt.% to about 45 wt.%, about 40 wt.% to about 50 wt.%, about 45 wt.% to about 55 wt.%, about 50 wt.% to about 60 wt.%, about 55 wt.% to about 65 wt.%, about 60 wt.% to about 70 wt.% oxidized cellulose, or any range including and/or in between any two of these values.

[0072] Additionally or alternatively, in some embodiments, the oxidized cellulose and/or ORC may comprise a weight-average molecular weight of about 50,000 to about 1,000,000. Additionally or alternatively, in some embodiments, the oxidized cellulose may have a weight-average molecular weight of about 50,000, about 60,000, about 75,000, about 100,000, about 150,000, about 200,000, about 250,000, about 300,000, about 400,000, about 500,000, about 600,000, about 700,000, about 800,000, about 900,000, about 1,000,000, or any range including and/or in between any two of the preceding values. Additionally or alternatively, in some embodiments, the oxidized cellulose may have a weight-average molecular weight of about 50,000 to about 200,000, about 100,000 to about 250,000, about 200,000 to about 400,000, about 300,000 to about 500,000, about 400,000 to about 800,000, about 600,000 to about 1,000,000, or any range including and/or in between any two of these values.

[0073] In any embodiment disclosed herein, 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. Additionally or alternatively, in some embodiments, the ORC may include fiber lengths of about 5 pm, about 10 pm, about 20 pm, about 50 pm, about 75 pm, about 100 pm, about 150 pm, about 200 pm, about 300 pm, about 400 pm, about 500 pm, about 600 pm, about 800 pm, about 1,000 pm, or any range including and/or in between any two of the preceding values. Additionally or alternatively, in some embodiments, the ORC may include fiber lengths of about 5 pm to about 50 pm, about 10 pm to about 100 pm, about 50 pm to about 250 pm, about 100 pm to about 500 pm, about 200 pm to about 600 pm, about 500 pm to about 1000 pm, or any range including and/or in between any two of the preceding values.

[0074] In any embodiment disclosed herein, the wound dressing compositions of the present technology may comprise about 30 wt.% to about 90 wt.% of the mixture of collagen and oxidized cellulose. Additionally or alternatively, in some embodiments, the collagen and oxidized cellulose mixture may be comprised at about 30 wt.%, about 40 wt.%, about 50 wt.%, about 60 wt.%, about 70 wt.%, about 80 wt.%, about 90 wt.%, or any range including and/or in between any two of the preceding values. In any embodiment disclosed herein, the mixture of collagen and oxidized cellulose can be included within the wound dressing compositions of the present technology at about 30 wt.% to about 40 wt.%, about 35 wt.% to about 45 wt.%, about 40 wt.% to about 50 wt.%, about 45 wt.% to about 55 wt.%, about 50 wt.% to about 60 wt.%, about 55 wt.% to about 65 wt.%, about 60 wt.% to about 70 wt.% about 65 wt.% to about 75 wt.%, about 70 wt.% to about 80 wt.%, about 75 wt.% to about 85 wt.%, about 80 wt.% to about 90 wt.%, or any range including and/or in between any two of these values.

[0075] In any embodiment disclosed herein, the first layer may be frozen at a temperature between about 0 °C to about -80 °C. Additionally or alternatively, in some embodiments, the first layer is frozen at a temperature of about 0 °C, about -5 °C, about -10 °C, about -15 °C, about -20 °C, about -30 °C, about -40 °C, about -50 °C, about -60 °C, about -70 °C, about -80 °C, or any range including and/or in between any two of these values. Additionally or alternatively, in some embodiments, the first layer is frozen at a temperature between about 0 °C to about -20 °C, about -10 °C to about -30 °C, about -20 °C to about -40 °C, about -30 °C to about -50 °C, about -40 °C to about -60 °C, about -50 °C to about -70 °C, about -60 °C to about -80 °C, or any range including and/or in between any two of these values.

[0076] In any embodiment disclosed herein, the formulation may include a solvent and an effective amount of at least one agent selected from the group consisting of a growth factor, an antimicrobial agent, a peptide, a biopolymer, and any combination thereof. Additionally or alternatively, in some embodiments, the solvent may be an aqueous solvent or an organic solvent. Additionally or alternatively, in some embodiments, the aqueous solvent may include a dilute organic acid, for example, acetic acid, citric acid, or a combination thereof.

[0077] In any embodiment disclosed herein, the formulation may include one or more of a binder, a colorant, a deodorant agent, a filler, a plasticizer, a viscosity modifier, an anti-microbial, or any combination thereof.

[0078] In any embodiment disclosed herein, the binder may be one or more of hyaluronic acid, chitosan, heparin, alginate, cellulose, fibrin, gelatin, chondroitin sulfate, agarose, dextran, carrageenan, silk, poly(ethylene glycol), poly(vinyl alcohol), polycaprolactone, polyphosphazene, polyglycolic acid, rosin, lactose, sucrose, tapioca starch, polyvinylpyrrolidone, or any combination thereof. Additionally or alternatively, in some embodiments, the formulation may include about 1 % w/v to about 10 % w/v of the binder. Additionally or alternatively, in some embodiments, the formulation may include about 1.0 % w/v, about 2.0 % w/v, about 3.0 % w/v, about 4.0 % w/v, about 5.0 % w/v, about 6.0 % w/v, about 7.0 % w/v, about 8.0 % w/v, about 9.0 % w/v, about 10 % w/v, or any range including and/or in between any two of the preceding values, of the binder. Additionally or alternatively, in some embodiments, the formulation may include about 1.0 % w/v to about 2.5 % w/v, about 2.0 % w/v to about 4 % w/v, 3.0 % w/v to about 5.0 % w/v, about 4.0 % w/v to about 6 % w/v, 5.0 % w/v to about 7.0 % w/v, about 6.0 % w/v to about 8 % w/v, 7.0 % w/v to about 9.0 % w/v, about 8.0 % w/v to about 10 % w/v, or any range including and/or in between any two of the preceding values, of the binder.

[0079] In any embodiment disclosed herein, the colorant may be one or more of direct red 80, bromophenol blue, toluidine blue, a food coloring, Allura Red AC, Erythrosine, Tartrazine 5, Anthocyanins, Curcuma, Annatto, or any combination thereof. Additionally or alternatively, in some embodiments, the formulation may include about 0.1 % w/v to about 5 % w/v of the colorant. Additionally or alternatively, in some embodiments, the formulation may include about 1 % w/v to about 10 % w/v of the colorant. Additionally or alternatively, in some embodiments, the formulation may include about 0.1 % w/v, about 0.15 % w/v, about 0.20 % w/v, about 0.30 % w/v, about 0.50 % w/v, about 0.75 % w/v, about 1.0 % w/v, about 2.0 % w/v, about 2.5 % w/v, about 3.0 % w/v, about 4.0 % w/v, about 5.0 % w/v, or any range including and/or in between any two of the preceding values, of the colorant. Additionally or alternatively, in some embodiments, the formulation may include about 0.1 % w/v to about 0.5 % w/v, about 0.25 % w/v to about 1.25 % w/v, about 1 % w/v to about 3 % w/v, about 2.0 % w/v to about 4.0 % w/v, about 2.5 % w/v to about 5.0 % w/v, or any range including and/or in between any two of the preceding values, of the colorant.

[0080] In any embodiment disclosed herein, the deodorant agent may be one or more of zinc ricinoleate, triethyl citrate, Saccharomyces ferment, Arctium lappa root extract, Humulus lupulus extract, Thymus vulgaris, or any combination thereof. Additionally or alternatively, in some embodiments, the formulation may include about 0.01 % w/v to about 5 % w/v of the deodorant agent. Additionally or alternatively, in some embodiments, the formulation may include about 0.01 % w/v, about 0.025 % w/v, about 0.05 % w/v, about 0.075 % w/v, 0.1 % w/v, about 0.20 % w/v, about 0.30 % w/v, about 0.50 % w/v, about 0.75 % w/v, about 1.0 % w/v, about 2.0 % w/v, about 2.5 % w/v, about 3.0 % w/v, about 4.0 % w/v, about 5.0 % w/v, or any range including and/or in between any two of the preceding values, of the deodorant agent. Additionally or alternatively, in some embodiments, the formulation may include about 0.01 % w/v to about 0.05 % w/v, about 0.025 % w/v to about 0.125 % w/v, 0.1 % w/v to about 0.5 % w/v, about 0.25 % w/v to about 1.25 % w/v, about 1 % w/v to about 3 % w/v, about 2.0 % w/v to about 4.0 % w/v, about 2.5 % w/v to about 5.0 % w/v, or any range including and/or in between any two of the preceding values, of the deodorant agent.

[0081] In any embodiment disclosed herein, the fdler may be one or more of amorphous silica, mica, kaolin, titanium dioxide, or any combination thereof. Additionally or alternatively, in some embodiments, the formulation may include about 0.01 % w/v to about 10 % w/v of the fdler. Additionally or alternatively, in some embodiments, the formulation may include about 0.01 % w/v, about 0.025 % w/v, about 0.05 % w/v, about 0.075 % w/v, 0.1 % w/v, about 0.20 % w/v, about 0.30 % w/v, about 0.50 % w/v, about 0.75 % w/v, about 1.0 % w/v, about 2.0 % w/v, about 2.5 % w/v, about 3.0 % w/v, about 4.0 % w/v, about 5.0 % w/v, about 6.0 % w/v, about 7.0 % w/v, about 8.0 % w/v, about 9.0 % w/v, about 10.0 % w/v, or any range including and/or in between any two of the preceding values, of the fdler. Additionally or alternatively, in some embodiments, the formulation may include about 0.01 % w/v to about 0.05 % w/v, about 0.025 % w/v to about 0.125 % w/v, 0.1 % w/v to about 0.5 % w/v, about 0.25 % w/v to about 1.25 % w/v, about 1 % w/v to about 3 % w/v, about 2.0 % w/v to about 4.0 % w/v, about 2.5 % w/v to about 7.5 % w/v, about 3.0 % w/v to about 10.0 % w/v, or any range including and/or in between any two of the preceding values, of the fdler.

[0082] In any embodiment disclosed herein, the plasticizer may include at least one component selected from the group consisting of an acetylated monoglyceride, an alkyl citrate, methyl ricinoleate, and glycerol. Additionally or alternatively, in some embodiments, the alkyl citrate may be triethyl citrate, acetyl triethyl citrate, tributyl citrate, acetyl tributyl citrate, trioctyl citrate, acetyl trioctyl citrate, trihexyl citrate, acetyl trihexyl citrate, butyryl trihexyl citrate, trimethyl citrate, or any combination thereof. Additionally or alternatively, in some embodiments, the formulation may include about 1 % w/v to about 10 % w/v of the at least one plasticizer. Thus, the at least one plasticizer may be included in an amount of about 0.01 % w/v, about 0.025 % w/v, about 0.05 % w/v, about 0.075 % w/v, 0.1 % w/v, about 0.20 % w/v, about 0.30 % w/v, about 0.50 % w/v, about 0.75 % w/v, about 1.0 % w/v, about 2.0 % w/v, about 2.5 % w/v, about 3.0 % w/v, about 4.0 % w/v, about 5.0 % w/v, about 6.0 % w/v, about 7.0 % w/v, about 8.0 % w/v, about 9.0 % w/v, about 10.0 % w/v, or any range including and/or in between any two of the preceding values, of the at least one plasticizer. Additionally or alternatively, in some embodiments, the formulation may include about 0.01 % w/v to about 0.05 % w/v, about 0.025 % w/v to about 0.125 % w/v, 0.1 % w/v to about 0.5 % w/v, about 0.25 % w/v to about 1.25 % w/v, about 1 % w/v to about 3 % w/v, about 2.0 % w/v to about 4.0 % w/v, about 2.5 % w/v to about 7.5 % w/v, about 3.0 % w/v to about 10.0 % w/v, or any range including and/or in between any two of the preceding values, of at least one plasticizer.

[0083] In any embodiment disclosed herein, the viscosity modifier may be one or more of sodium polyacrylate, acrylates/alkyl methacrylate block copolymers, polyacrylic acid, cellulose gum, hydroxyethylcellulose, stearyl alcohol, carbomer, or any combination thereof. Additionally or alternatively, in some embodiments, the formulation may include about 0.1 % w/v to about 10 % w/v of the viscosity modifier. Additionally or alternatively, in some embodiments, the formulation may include an amount of about 0.01 % w/v, about 0.025 % w/v, about 0.05 % w/v, about 0.075 % w/v, 0.1 % w/v, about 0.20 % w/v, about 0.30 % w/v, about 0.50 % w/v, about 0.75 % w/v, about 1.0 % w/v, about 2.0 % w/v, about 2.5 % w/v, about 3.0 % w/v, about 4.0 % w/v, about 5.0 % w/v, about 6.0 % w/v, about 7.0 % w/v, about 8.0 % w/v, about 9.0 % w/v, about 10.0 % w/v, or any range including and/or in between any two of the preceding values, of the viscosity modifier. Additionally or alternatively, in some embodiments, the formulation may include about 0.01 % w/v to about 0.05 % w/v, about 0.025 % w/v to about 0.125 % w/v, 0.1 % w/v to about 0.5 % w/v, about 0.25 % w/v to about 1.25 % w/v, about 1 % w/v to about 3 % w/v, about 2.0 % w/v to about 4.0 % w/v, about 2.5 % w/v to about 7.5 % w/v, about 3.0 % w/v to about 10.0 % w/v, or any range including and/or in between any two of the preceding values, of the viscosity modifier.

[0084] In any embodiment disclosed herein, the formulation may include about 0.01 wt.% to about 40 wt.%, or about 0.5 wt.% to about 5 wt.% of the at least one agent. Additionally or alternatively, in some embodiments, the formulation may include at least one agent at about 0.01 wt.%, about 0.025 wt.%, about 0.05 wt.%, about 0.1 wt.%, about 0.25 wt.%, about 0.5 wt.%, about 1 wt.%, about 5 wt.%, about 10 wt.%, about 15 wt.%, about 20 wt.%, about 25 wt.%, about 30 wt.%, about 35 wt.%, about 40 wt.%, or any range including and/or in between any two of these values. Additionally or alternatively, in some embodiments, the at least one agent may be included at about 0.01 wt.% to about 0.1 wt.%, about 0.05 wt.% to about 0.5 wt.%, about 0.1 wt.% to about 1 wt.%, about 0.5 wt.% to about 5 wt.%, about 1 wt.% to about 10 wt.%, about 5 wt.% to about 25 wt.%, about 10 wt.% to about 40 wt.% of the formulation, or any range including and/or in between any two of these values.

[0085] In any embodiment disclosed herein, the growth factor may include one or more of epidermal growth factor (EGF), hepatocyte growth factor (HGF), fibroblast growth factors (FGFs), or platelet-derived growth factor (PDGF). Additionally or alternatively, in some embodiments, the fibroblast growth factors comprise one or more of 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 any combination thereof.

[0086] In any embodiment disclosed herein, the antimicrobial agent may include one or more of polyhexamethylene biguanide, povidone-iodine, acetic acid, benzoic acid, povidone iodine, natamycin, nisin, citric acid, sorbic acid, propionic acid, honey, a silver compound, sulfites, or any combination thereof.

[0087] In any embodiment disclosed herein, the peptide may be one or more of defensins, histatins, cathelicidin FF-37, or any combination thereof.

[0088] In any embodiment disclosed herein, the biopolymer may be one or more of hyaluronic acid, fibrinogen, a chitosan, a polysaccharide, a gelatin, an oxidized cellulose, an oxidized regenerated cellulose (ORC), or any combination thereof. Additionally or alternatively, in some embodiments, the wound dressing composition of the present technology may include one or more synthetic polymers including, but not limited to, polyethylene glycol), poly(lactic acid), poly(vinyl alcohol) and polycaprolactone .

[0089] In any embodiment disclosed herein, the first layer has a thickness of about 1 mm to about 15 mm, or about 3 mm to about 8 mm. Additionally or alternatively, in some embodiments of the wound dressing disclosed herein, the first layer may have a thickness of about 1 mm, about 1.5 mm, about 2 mm, about 2.5 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, or any range including and/or in between any two of the preceding values. Additionally or alternatively, in some embodiments of the wound dressing disclosed herein, the first layer may have a thickness of about 1 mm to about 3 mm, about 2 mm to about 4 mm, about 3 mm to about 5 mm, about 4 mm to about 6 mm, about 5 mm to about 7 mm, about 6 mm to about 8 mm, about 7 mm to about 9 mm, about 8 mm to about 10 mm, about 9 mm to about 11 mm, about 10 mm to about 13 mm, about 12 mm to about 15 mm, or any range including and/or in between any two of the preceding values.

[0090] In any embodiment disclosed herein, the first layer and/or coated layer may optionally include a silver compound. Additionally or alternatively, in some embodiments, the first layer and/or coated layer may comprise about 0.1 wt.% to about 3 wt.% of a silver compound. Additionally or alternatively, in some embodiments, the first layer and/or coated layer may comprise about 0.1 wt.%, about 0. 25 wt.%, about 0.5 wt.%, about 0.75 wt.%, about 1 wt.%, about 1.5 wt.%, about 2 wt.%, about 2.5 wt.%, about 3 wt.%, or any range including and/or in between any two of the preceding values, of the silver compound. Additionally or alternatively, in some embodiments, the first layer and/or coated layer may comprise 0.1 wt.%, to about 0.3 wt.%, about 0.25 wt.% to about 0.75 wt.%, about 0.5 wt.% to about 1.5 wt.%, about 1 wt.% to about 2 wt.%, about 1.5 wt.% to about 3 wt.%, or any range including and/or in between any two of the preceding values, of the silver compound.

[0091] In any embodiment disclosed herein, the silver compound of the first and/or coated layer may include one or more pharmaceutically acceptable silver salts. Exemplary sources of the one or more pharmaceutically acceptable silver salts 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 p-aminobenzoate, silver p-aminosalicylate, nanocrystalline silver, any pharmaceutically acceptable salt thereof, or any combination thereof. Additionally or alternatively, in some embodiments of the wound dressing disclosed herein, the silver compound comprises a silver (II) oxide, silver (III) oxide, a silver oxy-salt, or any combination thereof. Additionally or alternatively, in some embodiments of the wound dressing disclosed herein, the silver oxy-salt may comprise a general formula of Ag(Ag30a)X, wherein X can include, but is not limited to, one or more acid anions such as sulfates, chlorides, phosphates, carbonates, citrates, tartrates, or oxalates; and wherein a is at least two.

[0092] In any embodiment disclosed herein, the first layer may include about 1 wt.% to about 15 wt.% of at least one plasticizer. Additionally or alternatively, in some embodiments, the at least one plasticizer of the first layer may be include at about 1 wt.%, about 2 wt.%, about 3 wt.%, about 4 wt.%, about 5 wt.%, about 6 wt.%, about 8 wt.%, about 10 wt.%, about 12.5 wt.%, about 15 wt.%, or any range including and/or in between any two of the preceding values. Additionally or alternatively, in some embodiments, the at least one plasticizer of the first layer may be include at about 1 wt.% to about 3 wt.%, about 2 wt.% to about 4 wt.%, about 3 wt.% to about 5 wt.%, about 4 wt.% to about 6 wt.%, about 5 wt. % to about 10 wt. %, about 8 wt. % to about 15 wt. %, or any range including and/or in between any two of the preceding values. Additionally or alternatively, in some embodiments, the at least one plasticizer may be an acetylated monoglyceride, an alkyl citrate, methyl ricinoleate, glycerol, polyvinylpyrrolidone, and any combination thereof. Additionally or alternatively, in some embodiments, the alkyl citrate may be triethyl citrate, acetyl triethyl citrate, tributyl citrate, acetyl tributyl citrate, trioctyl citrate, acetyl trioctyl citrate, trihexyl citrate, acetyl trihexyl citrate, butyryl trihexyl citrate, trimethyl citrate, and any combination thereof.

[0093] In any embodiment of the wound dressing disclosed herein, the anthocyanins are selected from the group consisting of cyanidin, delphinidin, malvidin, pelargonidin, peonidin, petunidin, and mixtures thereof.

[0094] In any embodiment of the wound dressing disclosed herein, the flavanols are selected from the group consisting of catechin, epicatechin, theaflavin, thearubigins, gallocatechin, epigallocatechin, or any gallate ester thereof, and mixtures thereof.

[0095] In any embodiment of the wound dressing disclosed herein, the flavanones are selected from the group consisting of eriodictyol, hesperetin, naringenin, and mixtures thereof.

[0096] In any embodiment of the wound dressing disclosed herein, the flavones are selected from the group consisting of apigenin, luteolin, tangeritin, and mixtures thereof.

[0097] In any embodiment of the wound dressing disclosed herein, the flavonols are selected from the group consisting of isorhamnetin, kaempferol, myricetin, proanthocyanidins, quercetin, rutin, and mixtures thereof.

[0098] In any embodiment of the wound dressing disclosed herein, the isoflavone phytoestrogens are selected from the group consisting of daidzein, genistein, glycitein, and any combination thereof.

[0099] In any embodiment of the wound dressing disclosed herein, the phenolic acids are selected from the group consisting of chicoric acid, chlorogenic acid, cinnamic acid, ellagic acid, ellagitannins, gallic acid, gallotannins, rosmarinic acid, salicylic acid, or any ester thereof, and any combination thereof.

[00100] In any embodiment of the wound dressing disclosed herein, the stillbenoids are selected from the group consisting of resveratrol, pterostilbene, and any combination thereof.

[00101] In any embodiment disclosed herein, the solid content of the first layer may be comprised at about 0.1 wt.%to about 10 wt.%. Additionally or alternatively, in some embodiments, the solid content of the first layer may be comprised at about 0.1 wt.%, about 0. 25 wt.%, about 0.5 wt.%, about 0.75 wt.%, about 1 wt.%, about 1.5 wt.%, about 2 wt.%, about 2.5 wt.%, about 3 wt.%, about 5 wt.%, about 7.5 wt.%, about 10 wt.%, or any range including and/or in between any two of the preceding values. Additionally or alternatively, in some embodiments, the solid content of the first layer may be comprised at about 0.1 wt.%, to about 0.3 wt.%, about 0.25 wt.% to about 0.75 wt.%, about 0.5 wt.% to about 1.5 wt.%, about 1 wt.% to about 3 wt.%, about 2 wt.% to about 6 wt.%, about 3 wt.% to about 10 wt.%, or any range including and/or in between any two of the preceding values.

[00102] In one aspect, the present disclosure provides an apparatus comprising a layer with a wound facing surface and an environmental-facing surface, wherein the freeze-dried layer comprises a first layer comprising a combination of collagen and an oxidized cellulose; and a coating forming a coated layer on the first layer, the coated layer comprising a formulation comprising an effective amount of at least one biologically active agent wherein the wound dressing composition does not visually appear to be wrinkled. Additionally or alternatively, in some embodiments, the formulation is applied to the first layer before freeze drying the first layer. Additionally or alternatively, in some embodiments, the coated layer has an average thickness of about 0.1 pm to about 1,000 pm. Additionally or alternatively, in some embodiments, the formulation is applied at a temperature of about 40 °C to about 80 °C. Additionally or alternatively, in some embodiments, the apparatus is resistant to structural changes compared to a control apparatus dressing that includes a collagen and an oxidized cellulose that has been freeze-dried before coating with the same formulation.

[00103] In one aspect, the present disclosure provides a device for the treatment of wound comprising a layer a layer with a wound-facing surface and an environmental-facing surface, wherein the freeze-dried layer comprises a first layer comprising a combination of collagen and an oxidized cellulose; and a coating forming a coated layer on the first layer, the coated layer comprising a formulation comprising an effective amount of at least one biologically active agent wherein the wound dressing composition does not visually appear to be wrinkled. Additionally or alternatively, in some embodiments, the formulation is applied to the first layer before freeze drying the first layer. Additionally or alternatively, in some embodiments, the coated layer has an average thickness of about 0.1 pm to about 1,000 pm. Additionally or alternatively, in some embodiments, the formulation is applied at a temperature of about 40 °C to about 80 °C. Additionally or alternatively, in some embodiments, the device is resistant to structural changes compared to a control device that includes a collagen and an oxidized cellulose that has been freeze-dried before coating with the same formulation.

Therapeutic and Prophylactic Methods of the Present Technology

[00104] In one aspect, the present disclosure provides a method for treating a wound in a subject in need thereof, the method including administering to the wound a wound dressing composition, apparatus, device, and/or layer of any embodiment disclosed herein. Additionally or alternatively in some embodiments, the wound dressing composition, apparatus, device, and/or layer induces, enhances, or promotes healing in the subject. Additionally or alternatively, in some embodiments, the subject is human.

[00105] In another related aspect, the present disclosure provides a method for preventing or reducing the risk of wound infection in a subject in need thereof, the method including administering to the wound a wound dressing composition, apparatus, device, and/or layer of any embodiment disclosed herein. Additionally or alternatively in some embodiments, the wound dressing composition, apparatus, device, and/or layer induces, enhances, or promotes healing in the subject. Additionally or alternatively, in some embodiments, the subject is human.

[00106] In any embodiment disclosed herein, the wound is an acute wound selected from the group consisting of bums, skin grafts, and dehisced surgical wounds. Additionally or alternatively, in some embodiments of the methods disclosed herein, the wound is a chronic wound selected from the group consisting of infectious wounds, venous ulcers, arterial ulcers, decubitis ulcers and diabetic ulcers. [00107] Any method known to those in the art for administering a wound dressing composition, apparatus, device, and/or layer to an acute wound 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, apparatuses, devices, and/or layers to a subject in need thereof, suitably a human. In some embodiments, the wound dressing composition, apparatus, device, and/or layer is applied directly to the wound. When used in vivo for therapy, the one or more wound dressing compositions, apparatus, device, and/or layer 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, apparatus, device, and/or layer used.

[00108] 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, apparatus, device, and/or layer useful in the methods may be administered to a subject in need thereof by any number of well-known methods for administering wound dressings, apparatuses, devices, and/or layers.

[00109] In any embodiment disclosed herein, the wound dressing compositions, apparatuses, devices, and/or layers are 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. Additionally or alternatively, in some embodiments, the wound dressing compositions, apparatuses, devices, and/or layers are administered one, two, three, four, or five times per day. Additionally or alternatively, in some embodiments, the wound dressing compositions, apparatuses, devices, and/or layers are administered daily for one, two, three, four or five weeks. Additionally or alternatively, in some embodiments, the wound dressing compositions, apparatuses, devices, and/or layers are administered daily for less than 6 weeks. Additionally or alternatively, in some embodiments, the wound dressing compositions, apparatuses, devices, and/or layers are administered daily for 6 weeks or more. Additionally or alternatively, in some embodiments, the wound dressing compositions, apparatuses, devices, and/or layers are administered daily for 12 weeks or more. Additionally or alternatively, in some embodiments, the wound dressing compositions, apparatuses, devices, and/or layers are administered every day, every other day, every third day, every fourth day, every fifth day, or every sixth day. Additionally or alternatively, in some embodiments, the wound dressing compositions, apparatuses, devices, and/or layers are administered weekly, bi-weekly, tri-weekly, or monthly. Additionally or alternatively, in some embodiments, the wound dressing compositions, apparatuses, devices, and/or layers are administered for a period of one, two, three, four, or five weeks. Additionally or alternatively, in some embodiments, the wound dressing compositions, apparatuses, devices, and/or layers are administered for six weeks or more. Additionally or alternatively, in some embodiments, the wound dressing compositions, apparatuses, devices, and/or layers are administered for twelve weeks or more. Additionally or alternatively, in some embodiments, the wound dressing compositions, apparatuses, devices, and/or layers are administered for a period of less than one year. Additionally or alternatively, in some embodiments, the wound dressing compositions, apparatuses, devices, and/or layers are administered for a period of more than one year.

[00110] In some embodiments, the wound dressing compositions apparatuses, devices, and/or layers can be changed for a chronic wound as appropriate. Additionally or alternatively, in some embodiments, the wound is a chronic wound selected from the group consisting of infectious wounds, venous ulcers, arterial ulcers, decubitis ulcers and diabetic ulcers.

[00111] In one aspect, the present disclosure provides a method of the treatment of a wound in a subject in need thereof, the method comprising administering to the subject the apparatus, wound dressings, device, and/or layer disclosed herein of any embodiment disclosed herein.

[00112] In one aspect, the present disclosure provides a method for preventing or reducing the risk of wound infection in a subject in need thereof, the method comprising administering to the subject the apparatus any embodiment disclosed herein.

Methods of Making the Wound Dressings of the Present Technology

[00113] Also disclosed herein are methods for making the wound dressings, apparatuses, devices, and/or layers of the present technology. In one aspect, the present disclosure provides a method for making a wound dressing composition, apparatus, and/or device of any embodiment disclosed herein, the method comprising, heating a formulation to about 40 °C to about 80 °C, depositing the formulation onto a first layer comprising a collagen and an oxidized cellulose to form a coated layer on the first layer, and freezing the coated layer; wherein the coated layer has an average thickness of about 0.1 pm to about 1,000 pm. Additionally or alternatively, in some embodiments, the method further comprises drying the coated layer, wound dressing composition, apparatus, and/or device.

[00114] In any embodiment disclosed herein, the method comprises heating the formulation to about 40 °C, about 45 °C, about 50 °C, about 55 °C, about 60 °C, about 65 °C, about 70 °C, about 75 °C, about 80 °C, or any range including and/or in between any two of these values. Additionally or alternatively, in some embodiments, the method comprises heating the formulation to about 40 °C to about 50 °C, about 45 °C to about 55 °C, about 50 °C to about 60 °C, about 55 °C to about 65 °C, about 60 °C to about 70 °C, about 65 °C to about 75 °C, about 70 °C to about 80 °C, or any range including and/or in between any two of these values.

[00115] In any embodiment disclosed herein, the formulation is heated for a period of time between about 0.5 min to about 240 min. Additionally or alternatively, in some embodiments, the formulation is heated for a period between about 0.5 min, about 1 min, about 2.5 min, about 5 min, about 7.5 min, about 10 min, about 15 min, about 30 min, about 45 min, about 60 min, about 90 min, about 120 min, about 150 min, about 180 min, about 210 min, about 240 min, or any range including and/or in between any two of these values. Additionally or alternatively, in some embodiments, the formulation is heated for a period between about 0.5 min to about 5 min, about 2 min to about 20 min, about 5 min to about 50 min, about 20 min to about 90 min, about 45 min to about 180 min, about 60 min to about 240 min, or any range including and/or in between any two of these values.

[00116] In any embodiment disclosed herein, the first layer may be frozen to form a frozen layer prior to deposition of the formulation, wherein the frozen layer optionally comprises a first surface. Additionally or alternatively, in some embodiments, the first surface of the frozen layer may be melted prior to deposition of the formulation.

[00117] In any embodiment disclosed herein, the first layer may be frozen at a temperature between about 0 °C to about -80 °C. Additionally or alternatively, in some embodiments, the first layer is frozen at a temperature of about 0 °C, about -5 °C, about -10 °C, about -15 °C, about -20 °C, about -30 °C, about -40 °C, about -50 °C, about -60 °C, about -70 °C, about -80 °C, or any range including and/or in between any two of these values. Additionally or alternatively, in some embodiments, the first layer is frozen at a temperature between about 0 °C to about -20 °C, about -10 °C to about -30 °C, about -20 °C to about -40 °C, about -30 °C to about -50 °C, about -40 °C to about -60 °C, about -50 °C to about -70 °C, about -60 °C to about -80 °C, or any range including and/or in between any two of these values. Additionally or alternatively, in some embodiments, the first layer may be frozen at a temperature lower than -80 °C. Additionally or alternatively, in some embodiments, the first layer may be frozen at or below the freezing temp of the mixture.

[00118] In any embodiment disclosed herein, the formulation may be deposited by a spray process or a thin layer application. Additionally or alternatively, in some embodiments, the thin layer application may be carried out by means of a brush.

[00119] In any embodiment disclosed herein, the coated layer may be frozen and/or freeze-dried for a period between about 0.1 min to about 10,000 min. Additionally or alternatively, in some embodiments, the coated layer may be frozen and/or freeze-dried for a period between about 0.1 min, about 0.5 min, about 1 min, about 5 min, about 15 min, about 30 min, about 60 min, about 90 min, about 120 min, about 200 min, about 500 min, about 750 min, about 1000 min, about 2000 min, about 4000 min, about 6000 min, about 8000 min, about 10,000 min, or any range including and/or in between any two of these values. Additionally or alternatively, in some embodiments, the coated layer may be frozen and/or freeze-dried for a period between about 0.1 min to about 1 min, about 0.5 min to about 5 min, about 2 min to about 20 min, about 5 min to about 50 min, about 20 min to about 200 min, about 50 min to about 500 min, about 200 min to about 2000 min, about 500 min to about 5000 min, about 1000 min to about 10000 min, or any range including and/or in between any two of these values. Additionally or alternatively, in some embodiments, the coated layer may be frozen and/or freeze-dried for at least 30 minutes. Additionally or alternatively, in some embodiments, the coated layer may be frozen and/or freeze-dried for at least 30 minutes.

[00120] In any embodiment disclosed herein, the coated layer is further dried and/or freeze-dried at a temperature above the temperature of the first drying and/or freeze drying of the coated fertilizer to facilitate removal of water. [00121] In some embodiments, the coated layer is freeze-dried for at least 30 min., 1 hour, 4 hours, 6 hours, 8 hours, 10 hours, 12 hours, 20 hours, or 24 hours.

[00122] For example, in some embodiments, the first layer may include a combination of a collagen and an oxidized regenerated cellulose (ORC) in a ratio of about 70 wt.%:30 wt.%, about 65 wt.%:35 wt.%, about 60 wt.%:40 wt.%, about 55 wt.%:45 wt.%, about 50 wt.%:50 wt.%, about 45 wt.%:55 wt.%, about 40 wt.%:60 wt.%, about 35 wt.%:65 wt.%, about 30 wt.%:70 wt.%, or any range including and/or in between any two of these values. Additionally or alternatively, in some embodiments, the first layer may include a combination of a collagen and an oxidized regenerated cellulose (ORC) in a ratio of about 55 wt.%: 45 wt.%. The first layer may be produced by adding a collagen (either in paste or powder form) to an acidic solution e.g., in order to swell the collagen. Examples of suitable acid solutions include, but are not limited to acetic acid and/or ascorbic acid. For example, the collagen may be contacted with the acid solution prior to forming an intermediate slurry with the ORC or an ORC- silver complex. In some embodiments, a slurry material may be generated with a solid content of about 0.1%, about 0.5%, about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 8%, about 10%, or any range including and/or in between any two of these values. Additionally or alternatively, in some embodiments, a slurry material may be generated with a solid content of about 0.1% to about 0.5%, about 0.4% to about 2%, about 1% to about 4%, about 3% to about 6%, about 4% to about 10%, or any range including and/or in between any two of these values. The resulting slurry is then frozen. In some embodiments, the resulting slurry may be flash frozen in a freezer at a temperature between about 0 °C, about -1 °C, about -5 °C, about -10 °C, about -15 °C, about -20 °C, about -30 °C, about -40 °C, about -50 °C, about -60 °C, about -70 °C, about -80 °C, or any range including and/or in between any two of these values. Additionally or alternatively, in some embodiments, the resulting slurry may be flash frozen in a freezer at a temperature between about 0 °C to about -10 °C, about -5 °C to about - 20 °C, about -10 °C to about -30 °C, about -20 °C to about -40 °C, about -30 °C to about -60 °C, about -50 °C to about -80 °C, or any range including and/or in between any two of these values. The first layer can then be coated with a thin layer application of a formulation at a temperature of about 40 °C, about 45 °C, about 50 °C, about 55 °C, about 60 °C, about 65 °C, about 70 °C, about 75 °C, about 80 °C, or any range including a temperature and/or range in between any two of these values. Additionally or alternatively, in some embodiments, the formulation may be applied at a temperature of about 40 °C to about 60 °C, about 50 °C to about 70 °C, about 60 °C to about 80 °C, or any range including a temperature and/or range in between any two of these values. The coated layer will then be freeze- dried at a reduced pressure to produce a wound dressing of the present technology.

[00123] In one aspect, the present disclosure provides a method of the treatment of a wound in a subject in need thereof, the method comprising administering to the subject the device for the treatment of wound of any embodiment. In one aspect, the present disclosure provides a method for preventing or reducing the risk of wound infection in a subject in need thereof, the method comprising administering to the subject the device for the treatment of wound of any embodiment. Additionally or alternatively, in some embodiments, the method further comprises performing negative pressure wound therapy to the wound prior to, concurrently with or after applying the wound dressing.

Kits Comprising the Wound Dressings of the Present Technology

[00124] 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 treating a wound in a subject in need thereof. The kit may optionally comprise components such as antiseptic wipes, ointment, adhesive tape, tweezers, or scissors.

[00125] Additionally or alternatively, in some embodiments, the kit, further comprises negative pressure wound therapy device. Additionally or alternatively, in some embodiments, the kit includes a patch, the patch comprising: a patch layer configured to prevent leaks, the patch layer comprising an opening, a patch backing layer adhered to the patch layer, and a patch delivery layer adhered to the patch layer, wherein the patch delivery layer comprises a slit, wherein the slit extends between an opening in the patch delivery layer and an exterior edge of the patch delivery layer; wherein placing the patch comprises positioning the opening in the patch layer over a vacuum port in the dressing layer and securing the patch layer to the dressing layer; and removing the patch delivery layer from the patch layer, wherein the slit permits the patch delivery layer to be removed when a fluid conduit is coupled to the vacuum port.

EXAMPLES

[00126] The present technology is further illustrated by the following Example, which should not be construed as limiting in any way. 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.

Example 1: Making the Wound Pres sins Compositions of the Present Technology

[00127] FIG. 1 provides a flow chart for making a wound dressing composition, device, layer, and/or apparatus, of the present technology. Briefly, a wound dressing composition, device, layer, and/or apparatus, of the present technology is made by freezing a slurry comprising collagen and oxidized regenerated cellulose (ORC), coating with a functional molecule, followed by freeze-drying (FIG. 1). The wound dressing composition, device, layer, and/or apparatus of the present technology may comprise a layer with a wound-facing surface and an environmental-facing surface, wherein the first layer comprises a collagen and an oxidized cellulose, and is coated with a formulation on the wound facing surface to form a coated layer on the first layer; wherein the coated layer has an average thickness of about 0.1 pm to about 1,000 pm, and wherein the formulation is applied at a temperature of about 40 °C to about 80 °C (FIG. 1), and wherein the wound dressing, device, layer, and/or apparatus is resistant to structural changes compared to a control dressing, device, layer, and/or apparatus that includes a collagen and an oxidized cellulose, and has been freeze-dried and then coated with the same formulation (FIGs. 1 and 3).

[00128] In a non-limiting example, the first layer comprises a combination of a collagen and an oxidized regenerated cellulose (ORC) at a ratio of 55 wt.%:45 wt.%. The first layer was produced by adding and homogenising oxidized regenerated cellulose (ORC), and collagen (either in paste or powder form) to an acidic solution (e.g., in order to swell the collagen), resulting in a slurry with 0.55 g solid collagen weight per 100 mL acid solution and 0.45 g ORC per 100 mL slurry. The slurry was then degassed and flash frozen in at -80°C in a freezer to form a block. After freezing a thin layer of a formulation or droplets of a formulation were applied to the wound-facing surface of the block by means of a brush or spraying, respectively, at a temperature of about 40°C. The resulting coated block was subsequently freeze-dried. FIG. 2 shows two non-limiting examples of wound dressings, devices, layers, and/or apparatuses made by this method (left panel shows a uniform coating of citric acid, right panel shows povidone -iodine dots coated on the first layer). The panels also show that surprisingly there is no visually apparent wrinkling of the surface due to the application of the formulation when compared to the uncoated area.

Example 2: Decreased Wrinkle Formation in the Wound Dressing Compositions of the Present Technology

[00129] To understand the effect of the sequence of (1) coating a dressing with a formulation, and (2) freeze drying on the texture of a wound dressing composition, the following two wound dressing compositions were prepared and compared with each other. In the first wound dressing composition, which is shown in FIG. 3 (left panel), the first layer was initially freeze-dried prior to applying a coating of a citric acid formulation. In the second wound dressing composition, which is shown FIG. 3 (right panel), the first layer was first coated with the citric acid formulation and then subjected to freeze- drying. As shown in FIG. 3 (left panel), the first wound dressing composition exhibited wrinkle formation and distortion. Surprisingly, the second wound dressing composition exhibited no visually detectable wrinkle formation or distortion, compared to the first wound dressing composition (FIG. 3 (right panel)). The second wound dressing is desirable because it allows close contact of the formulation with skin and/ or wound surface, ease of handling, ease of packaging, and a uniform dissolution rate. These data demonstrate that the sequence of (1) coating a dressing with a formulation, and (2) freeze can be used to obtain a desirable texture for a wound dressing composition. [00130] Accordingly, the wound dressing compositions of the present technology show reduced wrinkling and distortion, and are thus useful in the methods disclosed herein.

[00131] As shown in FIG. 2 the methods of making the wound dressing compositions of the present technology allow either uniform coating of formulations, or dotted/ patterned coating of formulations. The methods of making the wound dressing compositions of the present technology are amenable to coating the dressing with a formulation using processes such as soaking, imbibition, layering using an applicator, spray coating or continuous coating with a brush prior to freeze drying the wound dressing composition. This versatility enables coating of the wound dressing compositions of the present technology with a wider variety of formulations, including liquid, powder, semisolid or paste formulations. Likewise, aqueous or non-aqueous formulations can be easily applied.

[00132] Accordingly, the wound dressing compositions of the present technology are more versatile to make, and are thus useful in the methods disclosed herein.

Example 3: Improved Collagen Synthesis Upon Application of the Wound Dressing _. Compositions of the Present Technology

[00133] 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, devices, layers, and/or apparatuses of the present technology. Briefly, 8.4* 104 human fibroblasts (per well) are plated into 24-well plates, and then incubated at 37° C, 5% C02, 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 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 (CICP) present in the cell culture media. The level of CICP 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, devices, layers, and/or apparatuses of the present technology will exhibit increased collagen synthesis and improved wound healing of chronic or acute wounds compared to a standard collagen/ORC dressing.

[00134] Further, the collagen synthesis assay with dermal fibroblasts is performed with the wound dressing compositions devices, layers, and/or apparatuses of the present technology as well as a conventional coated wound dressing (e.g., a wound dressing that includes a collagen and an oxidized cellulose that has been freeze-dried first and then has been coated with the same formulation). Following the incubation of dermal fibroblast cells with SF-DMEM extracts of the samples, it is anticipated that the wound dressing compositions devices, layers, and/or apparatuses of the present technology exhibit increased collagen synthesis and improved wound healing of chronic or acute wounds compared to a conventional coated wound dressing. [00135] Accordingly, the dressings, devices, layers, and/or apparatuses 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, device, layer, and/or apparatuse of any embodiment disclosed herein.

Example 4: Wound Dressing _. Compositions of the Present Technology Decrease Protease Activity

[00136] A fluorometric assay is utilized to determine residual protease activities after incubation with the test wound dressing, device, layer, and/or apparatuse samples. Briefly, samples of the present technology, a standard collagen/ORC dressing, or a conventional coated wound dressing are incubated either in a solution of simulated wound fluid (SWF) containing human neutrophil elastase (HNE, 273 mU/mL) for 24 hours at 37° C, or in a solution of SWF containing matrix metalloproteinase-9 (MMP- 9, 1 pg/mL) for 2 hours at 37° C.

[00137] Following incubation, samples of the protease containing SWF are then evaluated by fluorometric assay to quantify residual protease activities. It is anticipated that the wound dressing compositions, devices, layers, and/or apparatuses of the present technology exhibit reduced protease levels and improved wound healing of chronic or acute wounds compared to standard collagen/ORC dressings and/or conventional coated wound dressings (e.g., a wound dressing that includes a collagen and an oxidized cellulose that has been freeze-dried first and then has been coated with the same formulation).

[00138] Accordingly, the wound dressing compositions, devices, layers, and/or apparatuses 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, device, layer, and/or apparatus of any embodiment disclosed herein.

EQUIVALENTS

[00139] 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. [00140] 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.

[00141] 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,” 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. 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.

[00142] 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.