WO2021152471A1

WO2021152471A1 - Visualization of wound protease levels

Info

Publication number: WO2021152471A1
Application number: PCT/IB2021/050617
Authority: WO
Inventors: Diwi ALLEN; Prathamesh Madhav KHARKAR
Original assignee: Kci Licensing, Inc.
Priority date: 2020-01-29
Filing date: 2021-01-27
Publication date: 2021-08-05

Abstract

The present disclosure relates generally to wound dressing compositions that include at least one type of microcapsule comprising one or more biocompatible markers. Such wound dressing compositions may be used to detect protease levels in a wound bed. Methods of using the wound dressing compositions for treating wounds and for selecting patients for a therapeutic regimen, as well as kits for use in practicing the methods are also provided.

Description

VISUALIZATION OF WOUND PROTEASE LEVELS

CROSS-REFERENCE TO RELATED APPLICATIONS [001] This application claims the benefit of priority to U.S. Provisional Application No. 62/967,144, filed on January 29, 2020, which is incorporated herein by reference in its entirety.

TECHNICAL FIELD

[002] The present technology relates generally to wound dressing compositions that include at least one type of microcapsule comprising a biocompatible marker. Such wound dressing compositions may be used to detect protease levels in a wound bed. Methods of using the wound dressing compositions for treating wounds and for selecting patients for a therapeutic regimen, as well as kits for use in practicing the methods are also provided.

BACKGROUND

[003] 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.

[004] A wide variety of materials and devices, generally characterized as “dressings,” are known in the art for use in treating an injury or other tissue disruptions. Such wounds may be the result of trauma, surgery, or disease, and may affect skin or other tissues. Dressings may control bleeding, absorb wound exudate, ease pain, assist in debriding the wound, protect wound tissue from infection, or otherwise promote healing and protect the wound from further damage.

SUMMARY OF THE PRESENT TECHNOLOGY [005] In one aspect, the present disclosure provides a wound dressing composition comprising a substrate sheet; and a plurality of microcapsules comprising a biocompatible marker encapsulated by an encapsulating material, wherein the encapsulating material comprises a first collagen. Additionally or alternatively, in some embodiments, the wound dressing composition further comprises a wicking layer. Additionally or alternatively, in some embodiments, the wound dressing composition further comprises a sealing layer comprising a window for viewing the wicking layer and/or the substrate sheet.

[006] In one aspect, the present disclosure provides a wound dressing composition comprising a substrate sheet; and a plurality of microcapsules comprising a biocompatible marker encapsulated by a first encapsulating material, wherein the first encapsulating material comprises a gelatin and an oxidized regenerated cellulose (ORC), and a second encapsulating material, wherein the second encapsulating material comprises a collagen, wherein the second encapsulating material encapsulates the first encapsulating material. Additionally or alternatively, in some embodiments, the wound dressing composition further comprises a wicking layer. Additionally or alternatively, in some embodiments, the wound dressing composition further comprises a sealing layer comprising a window for viewing the wicking layer and/or the substrate sheet.

[007] In one aspect, the present disclosure provides a wound dressing composition comprising a substrate sheet; a plurality of first microcapsules comprising a first biocompatible marker encapsulated by a first encapsulating material, wherein the first encapsulating material comprises a first collagen; and a plurality of second microcapsules comprising a second biocompatible marker encapsulated by a second encapsulating material, wherein the second encapsulating material comprises a gelatin and an oxidized regenerated cellulose (ORC), and a third encapsulating material, wherein the third encapsulating material comprises a second collagen, wherein the third encapsulating material encapsulates the second encapsulating material. Additionally or alternatively, in some embodiments, the wound dressing composition further comprises a wicking layer. Additionally or alternatively, in some embodiments, the wound dressing composition further comprises a sealing layer comprising a window for viewing the wicking layer and/or the substrate sheet.

[008] Additionally, or alternatively, in some embodiments, the plurality of first microcapsules and the plurality of second microcapsules have a diameter of from about 0.01 pm to about 1000 pm. In some embodiments, the plurality of first microcapsules and the plurality of second microcapsules have a diameter in a range independently selected from the group consisting of from about 0.01 pm to about 0.1 pm, from about 0.05 pm to about 0.25 pm, from about 0.1 pm to about 1 pm, from about 0.5 pm to about 2.5 pm, 1 pm to about 10 pm, from about 5 pm to about 25 pm, from about 10 pm to about 100 pm, from about 50 pm to about 250 pm, from about 100 pm to about 1000 pm, and any range including and/or in between any two of these values. In some embodiments, the plurality of first microcapsules and the plurality of second microcapsules have a diameter of about 5 pm to about 200 pm.

[009] Additionally, or alternatively, in some embodiments, the plurality of first microcapsules and the plurality of second microcapsules have different diameters. In some embodiments, the plurality of first microcapsules are smaller than the plurality of second microcapsules.

[0010] Additionally, or alternatively, in some embodiments, the plurality of first microcapsules are digested faster by a matrix metalloproteinase (MMP) present in wound exudate compared to the plurality of second microcapsules.

[0011] Additionally, or alternatively, in some embodiments, the first collagen and the second collagen are independently recombinant collagen or naturally occurring collagen. Additionally, or alternatively, in some embodiments, the first collagen and the second collagen are mammalian collagens. In some embodiments, the first collagen and the second collagen independently comprise a bovine collagen, a human collagen, or a combination thereof. In some embodiments, the first collagen and/or second collagen comprises one or more of type I human recombinant collagen, type III human recombinant collagen, type I bovine collagen, type III bovine collagen, or a combination of any two or more thereof. [0012] Additionally, or alternatively, in some embodiments, the first biocompatible marker and the second biocompatible marker independently comprise a dye or a fluorochrome. Additionally, or alternatively, in some embodiments, the first biocompatible marker and the second biocompatible marker are water soluble. Additionally, or alternatively, in some embodiments, the first biocompatible marker and/or the second biocompatible marker is substantially invisible when viewed using a light having wavelength of from about 380 nm to 740 nm. Additionally, or alternatively, in some embodiments, the first biocompatible marker and/or the second biocompatible marker comprises a pH indicator.

[0013] Additionally, or alternatively, in some embodiments, the substrate sheet comprises comprise a wound-facing side and an environmental -facing side.

[0014] Additionally, or alternatively, in some embodiments, the plurality of first microcapsules and/or the plurality of second microcapsules are embedded in the substrate sheet. Additionally, or alternatively, in some embodiments, the plurality of first microcapsules and/or the plurality of second microcapsules are coupled to the wound-facing side of the substrate sheet. In some embodiments, the plurality of first microcapsules and/or the plurality of second microcapsules are coupled to the entire wound-facing side of the substrate sheet. Additionally, or alternatively, in some embodiments, the plurality of first microcapsules and/or the plurality of second microcapsules are coupled to only a part of the wound-facing side of the substrate sheet.

[0015] Additionally, or alternatively, in some embodiments, the wound dressing comprises about 0.01 wt.% to about 65 wt.%, more preferably about 0.01 wt % to about 10 wt%, of the plurality of first microcapsules. Additionally, or alternatively, in some embodiments, the wound dressing comprises about 0.01 wt.% to about 65 wt.%, more preferably about 0.01 wt % to about 10 wt%, of the plurality of second microcapsules.

[0016] Additionally, or alternatively, in some embodiments, the plurality of first microcapsules, the plurality of second microcapsules and/or the substrate sheet comprises a silver compound. In some embodiments, the plurality of first microcapsules, the plurality of second microcapsules and/or the substrate sheet comprises about 0.1 wt.% to about 3 wt.% of the silver compound. In some embodiments, the silver compound comprises one or more pharmaceutically acceptable silver salts.

In some embodiments, the one or more pharmaceutically acceptable silver salts is selected from the group consisting of silver oxide, silver oxysalts, silver chromate, silver allantoinate, silver borate, silver glycerolate, silver nitrate, silver acetate, silver chloride, silver sulfate, silver lactate, silver bromide, silver iodide, silver carbonate, silver citrate, silver laurate, silver deoxycholate, silver salicylate, silver p-aminobenzoate, silver p-aminosalicylate, nanocrystalline silver, any pharmaceutically acceptable salt thereof, and a combination any two or more thereof.

[0017] Additionally, or alternatively, in some embodiments, the substrate sheet further comprises one or more additional additives selected from the group consisting of an antimicrobial agent, an antioxidant, a signaling protein, and a combination any two or more thereof. [0018] Additionally, or alternatively, in some embodiments, the plurality of first microcapsules, the plurality of second microcapsules and/or the substrate sheet comprises about 0.001 wt.% to about 5 wt.% of the antimicrobial agent. In some embodiments, the antimicrobial agent comprises one or more of tetracycline, penicillins, terramycins, erythromycin, bacitracin, neomycin, polymycin B, mupirocin, clindamycin, colloidal silver, silver sulfadiazine, chlorhexidine, povidone iodine, triclosan, sucralfate, quaternary ammonium salts, pharmaceutically acceptable silver salts, or a combination any two or more thereof.

[0019] Additionally, or alternatively, in some embodiments, the plurality of first microcapsules, the plurality of second microcapsules and/or the substrate sheet comprises about 0.001 wt.% to about 5 wt.% of the antioxidant. In some embodiments, the antioxidant comprises one or more of anthocyanins, astaxanthin, bilirubin, canthaxanthin, capsaicin, citric acid, curcumin, coenzyme Q10, eugenol, flavanols, flavonolignans, flavanones, flavones, flavonols, iodide, isoflavone phytoestrogens, lutein, lycopene, manganese, melatonin, N-acetylcysteine, oxalic acid, phenolic acids, phytic acid, R- a-lipoic acid, stilbenoids, tocopherol, tocotrienol, vitamin A, vitamin C, vitamin E, xanthones, zeaxanthin, a-carotene, b-carotene, or any combination thereof. In some embodiments, the anthocyanins are selected from the group consisting of cyanidin, delphinidin, malvidin, pelargonidin, peonidin, petunidin, and any combination thereof. In some embodiments, the flavanols are selected from the group consisting of catechin, epicatechin, theaflavin, thearubigins, gallocatechin, epigallocatechin, or any gallate ester thereof, and any combination thereof. In some embodiments, the flavanones are selected from the group consisting of eriodictyol, hesperetin, naringenin, and any combination thereof. In some embodiment, the flavones are selected from the group consisting of apigenin, luteolin, tangeritin, and any combination thereof. In some embodiments, the flavonols are selected from the group consisting of isorhamnetin, kaempferol, myricetin, proanthocyanidins, quercetin, rutin, and any combination thereof. In some embodiments, the isoflavone phytoestrogens are selected from the group consisting of daidzein, genistein, glycitein, and any combination thereof. In some embodiments, 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. In some embodiments, the stilbenoids are selected from the group consisting of resveratrol, pterostilbene, and any combination thereof.

[0020] Additionally, or alternatively, in some embodiments, the plurality of first microcapsules, the plurality of second microcapsules and/or the substrate sheet comprises about 0.001 wt.% to about 5 wt.% of the signaling protein. In some embodiments, the signaling protein comprises one or more of platelet-derived growth factor (PDGF), transforming growth factor beta (TGF ), fibroblast growth factors (FGFs), epidermal growth factor (EGF), or any combination thereof. In some embodiments, the fibroblast growth factors (FGFs) 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 a combination any two or more thereof.

[0021] In one aspect, the present disclosure provides an apparatus comprising a substrate sheet; a plurality of first microcapsules comprising a first biocompatible marker encapsulated by a first encapsulating material, wherein the first encapsulating material comprises a first collagen; and a plurality of second microcapsules comprising a second biocompatible marker encapsulated by a second encapsulating material, wherein the second encapsulating material comprises a gelatin and an ORC, and a third encapsulating material, wherein the third encapsulating material comprises a second collagen, wherein the third encapsulating material encapsulates the second encapsulating material. Additionally or alternatively, in some embodiments, the apparatus further comprises a wicking layer. Additionally or alternatively, in some embodiments, the apparatus further comprises a sealing layer comprising a window for viewing the wicking layer and/or the substrate sheet.

[0022] In one aspect, the present disclosure provides a system comprising a negative pressure source; and a wound dressing of any embodiment disclosed herein, or an apparatus of any embodiment disclosed herein.

[0023] In one aspect, the present disclosure provides a wound dressing composition comprising a wound-facing sheet, a substrate sheet; and a plurality of microcapsules comprising a biocompatible marker encapsulated by an encapsulating material, wherein the encapsulating material comprises a collagen, wherein the plurality of microcapsules are embedded in or coupled to the substrate sheet. Additionally or alternatively, in some embodiments, the substrate sheet has a wound-facing side and an environmental facing side. Additionally or alternatively, in some embodiments, the microcapsules are coupled to the environmental facing side of the substrate sheet. Additionally or alternatively, in some embodiments, the wound dressing composition further comprises a wicking layer. Additionally or alternatively, in some embodiments, the wound dressing composition further comprises a sealing layer comprising a window for viewing the wicking layer and/or the substrate sheet.

[0024] In one aspect, the present disclosure provides a method for treating a wound in a subject in need thereof, comprising administering the wound a dressing of any of the embodiments disclosed herein, the apparatus of any of the embodiments disclosed herein or the system of any of the embodiments disclosed herein to the wound. Additionally or alternatively, in some embodiments, the method further comprises applying a retainer layer and/or a foam over the wound dressing. Additionally or alternatively, in some embodiments, the method further comprises applying the drape over the wound dressing of any of the embodiments disclosed herein, the apparatus of any of the embodiments disclosed herein, or the system of any of the embodiments disclosed herein and/or the retainer layer, wherein the drape is configured to seal the wound dressing and/or the retainer layer and the wound site. Additionally or alternatively, in some embodiments, the method further comprises applying negative pressure to the wound, wherein the vacuum is configured to be fluidly connected to the drape through tubing.

[0025] In one aspect, the present disclosure provides a method for selecting a patient suspected of, or diagnosed as having an elevated level of inflammatory mediators in a wound for treatment with a therapeutic regimen comprising (a) administering a device to the wound, wherein the device comprises a wound dressing of any of the embodiments disclosed herein, a apparatus of any of the embodiments disclosed herein, or a system of any of the embodiments disclosed herein; (b) detecting change in color or fluorescence via a window for viewing the wicking layer and/or the substrate sheet; and (c) administering a therapeutic regimen when elevated levels of inflammatory mediators are detected compared to a predetermined threshold or a negative control sample. Additionally or alternatively, in some embodiments, the method further comprises applying the drape over the wound dressing and/or the retainer layer, wherein the drape is configured to seal the wound dressing of any of the embodiments disclosed herein, the apparatus of any of the embodiments disclosed herein or the system of any of the embodiments disclosed herein and/or the retainer layer and the wound site. Additionally or alternatively, in some embodiments, the method further comprises optionally a vacuum for applying negative pressure to the wound, wherein the vacuum is configured to be fluidly connected to the drape through tubing. In some embodiments, the therapeutic regimen is selected from the group consisting of change of dressing, administration of one or more antibiotics, hospitalization, or a combination thereof.

[0026] In one aspect, the present disclosure provides a method for making a wound dressing composition comprising: (a) providing (i) a substrate sheet; (ii) a plurality of first microcapsules comprising a first biocompatible marker encapsulated by a first encapsulating material, wherein the first encapsulating material comprises a first collagen; and (iii) a plurality of second microcapsules comprising a second biocompatible marker encapsulated by a second encapsulating material, wherein the second encapsulating material comprises a gelatin and an ORC, and a third encapsulating material, wherein the third encapsulating material comprises a second collagen, wherein the third encapsulating material encapsulates the second encapsulating material; (iv) optionally, a wicking layer; and (v) a sealing layer comprising a window for viewing the wicking layer and/or the substrate sheet; wherein the substrate sheet, the wicking layer, and the sealing layer independently comprise a wound-facing side and an environmental-facing side; (b) combining the substrate sheet, the wicking layer, and the sealing layer such that the wound-facing side of the sealing layer is coupled to the environmental- facing side of the wicking layer and/or the substrate sheet; and optionally, the wound-facing side of the wicking layer is coupled to the environmental-facing side of the substrate sheet; (c) coupling the plurality of first microcapsules with the wound-facing side of the substrate sheet; and (d) coupling the plurality of second microcapsules with the wound-facing side of the substrate sheet.

[0027] Additionally, or alternatively, in some embodiments, the plurality of first microcapsules and the plurality of second microcapsules have a diameter of from about 0.01 pm to about 1000 pm. In some embodiments, the plurality of first microcapsules and the plurality of second microcapsules have a diameter in a range independently selected from the group consisting of from about 0.01 pm to about 0.1 pm, from about 0.05 pm to about 0.25 pm, from about 0.1 pm to about 1 pm, from about 0.5 pm to about 2.5 pm, 1 pm to about 10 pm, from about 5 pm to about 25 pm, from about 10 pm to about 100 pm, from about 50 pm to about 250 pm, from about 100 pm to about 1000 pm, or any range including and/or in between any two of these values. In some embodiments, the plurality of first microcapsules and the plurality of second microcapsules have a diameter of about 5 pm to about 200 pm.

[0028] Additionally, or alternatively, in some embodiments, the plurality of first microcapsules and the plurality of second microcapsules have different sizes. In some embodiments, the plurality of first microcapsules are smaller than the plurality of second microcapsules.

[0029] Additionally, or alternatively, in some embodiments, the plurality of first microcapsules are digested faster by a matrix metalloproteinase (MMP) present in wound exudate compared to the plurality of second microcapsules.

[0030] Additionally, or alternatively, in some embodiments, the first collagen and the second collagen are independently recombinant or naturally occurring. Additionally, or alternatively, in some embodiments, the first collagen and the second collagen are mammalian collagens. In some embodiments, the first collagen and the second collagen independently comprise a bovine collagen, a human collagen, or a combination thereof. In some embodiments, the first collagen and/or second collagen comprises one or more of type I human recombinant collagen, type III human recombinant collagen, type I bovine collagen, type III bovine collagen, or a combination of any two or more thereof.

[0031] Additionally, or alternatively, in some embodiments, the first biocompatible marker and the second biocompatible marker independently comprise a dye or a fluorochrome. Additionally, or alternatively, in some embodiments, the first biocompatible marker and the second biocompatible marker are water soluble. Additionally, or alternatively, in some embodiments, the first biocompatible marker and/or the second biocompatible marker is substantially invisible when viewed using a light having wavelength of from about 380 nm to 740 nm. Additionally, or alternatively, in some embodiments, the first biocompatible marker and/or the second biocompatible marker is a pH indicator.

[0032] Additionally, or alternatively, in some embodiments, the plurality of first microcapsules and/or the plurality of second microcapsules are embedded in the substrate sheet. Additionally, or alternatively, in some embodiments, the substrate sheet comprises comprise a wound-facing side and an environmental-facing side. Additionally, or alternatively, in some embodiments, the plurality of first microcapsules and/or the plurality of second microcapsules are coupled to the wound-facing side of the substrate sheet. In some embodiments, the plurality of first microcapsules and/or the plurality of second microcapsules are coupled to the entire wound-facing side of the substrate sheet. Additionally, or alternatively, in some embodiments, the plurality of first microcapsules and/or the plurality of second microcapsules are coupled to only a part of the wound-facing side of the substrate sheet.

[0033] Additionally, or alternatively, in some embodiments, the wound dressing comprises about 0.01 wt.% to about 65 wt.%, more preferably about 0.01 wt % to about 10 wt%, of the plurality of first microcapsules. Additionally, or alternatively, in some embodiments, the wound dressing comprises about 0.01 wt.% to about 65 wt.%, more preferably about 0.01 wt % to about 10 wt%, of the plurality of second microcapsules.

[0034] Additionally, or alternatively, in some embodiments, the plurality of first microcapsules, the plurality of second microcapsules and/or the substrate sheet comprises a silver compound. In some embodiments, the plurality of first microcapsules, the plurality of second microcapsules and/or the substrate sheet comprises about 0.1 wt.% to about 3 wt.% of the silver compound. In some embodiments, the silver compound comprises one or more pharmaceutically acceptable silver salts.

In some embodiments, the one or more pharmaceutically acceptable silver salts is selected from the group consisting of silver oxide, silver oxysalts, silver chromate, silver allantoinate, silver borate, silver glycerolate, silver nitrate, silver acetate, silver chloride, silver sulfate, silver lactate, silver bromide, silver iodide, silver carbonate, silver citrate, silver laurate, silver deoxycholate, silver salicylate, silver p-aminobenzoate, silver p-aminosalicylate, nanocrystalline silver, any pharmaceutically acceptable salt thereof, and any combination thereof.

[0035] Additionally, or alternatively, in some embodiments, the plurality of first microcapsules, the plurality of second microcapsules and/or the substrate sheet comprises further comprises one or more additional additives selected from the group consisting of an antimicrobial agent, an antioxidant, a signaling protein, or a combination any two or more thereof.

[0036] Additionally, or alternatively, in some embodiments, the plurality of first microcapsules, the plurality of second microcapsules and/or the substrate sheet comprises about 0.001 wt.% to about 5 wt.% of the antimicrobial agent. In some embodiments, the antimicrobial agent comprises one or more of tetracycline, penicillins, terramycins, erythromycin, bacitracin, neomycin, polymycin B, mupirocin, clindamycin, colloidal silver, silver sulfadiazine, chlorhexidine, povidone iodine, triclosan, sucralfate, quaternary ammonium salts, pharmaceutically acceptable silver salts, or any combination thereof.

[0037] Additionally, or alternatively, in some embodiments, the plurality of first microcapsules, the plurality of second microcapsules and/or the substrate sheet comprises about 0.001 wt.% to about 5 wt.% of the antioxidant. In some embodiments, the antioxidant comprises one or more of anthocyanins, astaxanthin, bilirubin, canthaxanthin, capsaicin, citric acid, curcumin, coenzyme Q10, eugenol, flavanols, flavonolignans, flavanones, flavones, flavonols, iodide, isoflavone phytoestrogens, lutein, lycopene, manganese, melatonin, N-acetylcysteine, oxalic acid, phenolic acids, phytic acid, R- a-lipoic acid, stilbenoids, tocopherol, tocotrienol, vitamin A, vitamin C, vitamin E, xanthones, zeaxanthin, a-carotene, b-carotene, or any combination thereof. In some embodiments, the anthocyanins are selected from the group consisting of cyanidin, delphinidin, malvidin, pelargonidin, peonidin, petunidin, and any combination thereof. In some embodiments, the flavanols are selected from the group consisting of catechin, epicatechin, theaflavin, thearubigins, gallocatechin, epigallocatechin, or any gallate ester thereof, and any combination thereof. In some embodiments, the flavanones are selected from the group consisting of eriodictyol, hesperetin, naringenin, and any combination thereof. In some embodiment, the flavones are selected from the group consisting of apigenin, luteolin, tangeritin, and any combination thereof. In some embodiments, the flavonols are selected from the group consisting of isorhamnetin, kaempferol, myricetin, proanthocyanidins, quercetin, rutin, and any combination thereof. In some embodiments, the isoflavone phytoestrogens are selected from the group consisting of daidzein, genistein, glycitein, and any combination thereof.

In some embodiments, 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. In some embodiments, the stilbenoids are selected from the group consisting of resveratrol, pterostilbene, and any combination thereof.

[0038] Additionally, or alternatively, in some embodiments, the plurality of first microcapsules, the plurality of second microcapsules and/or the substrate sheet comprises about 0.001 wt.% to about 5 wt.% of the signaling protein. In some embodiments, the signaling protein comprises one or more of platelet-derived growth factor (PDGF), transforming growth factor beta (TGF ), fibroblast growth factors (FGFs), epidermal growth factor (EGF), or any combination thereof. In some embodiments, the fibroblast growth factors (FGFs) 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. [0039] Also provided herein are kits comprising the wound dressings of any embodiment disclosed herein and instructions for use.

BRIEF DESCRIPTION OF THE DRAWINGS [0040] FIG. 1A shows a diagrammatic representation of a non-limiting embodiment of a first microcapsule of the present technology. In this embodiment, a colored pigment is encapsulated with collagen.

[0041] FIG. IB shows a diagrammatic representation of a non-limiting embodiment of a second microcapsule of the present technology. In this embodiment, a colored pigment is encapsulated with gelatin, and is further encapsulated with collagen.

[0042] FIG. 2 shows a diagrammatic representation of a wound dressing composition of the present technology. In this embodiment, the first and the second microcapsules of the present technology are embedded in a substrate sheet of a wound dressing composition.

[0043] FIG. 3 shows an embodiment wherein the first and the second microcapsules of the present technology are embedded in or bound to a patch of the film / adhesive layer of a T.R.A.C.™ Pad. [0044] FIG. 4 shows a diagrammatic representation of a wound dressing composition of the present technology. In this embodiment, the first and the second microcapsules of the present technology are bound to a small patch. This embodiment may include a window which can be used to view whether the microcapsules are degraded.

DETAILED DESCRIPTION

[0045] 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.

[0046] The present disclosure is directed to wound dressing compositions that include substrate sheet, and at least one type of the microcapsules that includes at least one biocompatible marker. The wound dressings of the present technology advantageously allow detection of elevated levels of inflammation mediators such as proteases, including the matrix metallopeptidases (MMPs) that are elevated in wound during inflammation, thereby allowing therapeutic intervention, if needed.

[0047] In one aspect, the present disclosure provides a wound dressing composition comprising a substrate sheet; a plurality of first microcapsules comprising a first biocompatible marker encapsulated by a first encapsulating material, wherein the first encapsulating material comprises a first collagen; and a plurality of second microcapsules comprising a second biocompatible marker encapsulated by a second encapsulating material, wherein the second encapsulating material comprises a gelatin and an ORC, and a third encapsulating material, wherein the third encapsulating material comprises a second collagen, wherein the third encapsulating material encapsulates the second encapsulating material. Additionally or alternatively, in some embodiments, the wound dressing composition further comprises a wicking layer and/or a sealing layer comprising a window for viewing the wicking layer and/or the substrate sheet. [0048] FIG.1A provides representative, non-limiting illustration of an embodiment of the first microcapsule present in the wound dressings of the present technology.

[0049] FIG. IB provides representative, non-limiting illustration of an embodiment of the first microcapsule present in the wound dressings of the present technology.

[0050] FIGs.2-4 provides representative, non-limiting illustrations the wound dressings of the present technology.

Definitions

[0051] 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. [0052] 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.

[0053] As used herein, the terms “wound dressing composition” and “apparatus” are used interchangeably.

[0054] 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.

[0055] 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.

[0056] As used herein, the term “effective amount” refers to a quantity sufficient to achieve a desired therapeutic and/or prophylactic effect, e.g. , an amount which results in wound healing or a reduction of one or more signs or symptoms associated with a wound described herein. In the context of therapeutic and/or prophylactic applications, the wound dressing 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.

[0057] 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.

[0058] The term “mammalian recombinant collagen” refers to collagen manufactured by culturing a non-human organism or mammalian or non-mammalian cells to express at least one exogenous gene encoding a collagen in the culturing system. The term “human recombinant collagen” refers to collagen manufactured by culturing a non-human organism or mammalian or non-mammalian cells to express at least one human gene encoding a collagen. The human recombinant collagen may be selected from the group consisting of collagen type I, type II, type III, type IV, type V, type VI, type VII, type VIII, type IX, type X, type XI, type XII, type XIII, type XIV, type XV, type XVI, type XVII, type XVIII, type XIX, type XX, type XXI, type XXII, type XXIII, type XXIV, type XXV, type XXVI, and type XXVII. The human recombinant collagen can be collagen of one type free of any other type, or can be a mixture of collagen types. Suitably, the human recombinant collagen comprises collagens selected from the group consisting of collagen type I, collagen type III, and mixtures thereof. The term "bovine recombinant collagen" refers to collagen manufactured by culturing a non-human organism or mammalian or non-mammalian cells to express at least one bovine gene encoding a collagen. The bovine recombinant collagen may be selected from the group consisting of collagen type I, type II, type III, and type IV. The bovine recombinant collagen can be collagen of one type free of any other type, or can be a mixture of collagen types. Suitably, the bovine recombinant collagen comprises collagens selected from the group consisting of collagen type I, collagen type III, and mixtures thereof.

[0059] As understood by one of ordinary skill in the art, “molecular weight” (also known as “relative molar mass”) is a dimensionless quantity but is converted to molar mass by multiplying by 1 gram/mole - for example, collagen with a weight-average molecular weight of 5,000 has a weight- average molar mass of 5,000 g/mol.

[0060] As used herein, the term “NPWT” refers to negative pressure wound therapy, which is a type of wound therapy that involves applying negative pressure (relative to atmospheric pressure) to a wound bed to promote wound healing. Typically, a dressing is sealed over a wound site and air is pumped out of the dressing to create negative pressure at the wound site. In some NPWT systems, wound exudate and other fluid is pumped out of the dressing and collected by a canister.

[0061] “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.

[0062] 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 Dressings of the Present Technology [0063] 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.

[0064] Proteases play pivotal roles in normal wound healing processes. In general, different wound-related proteases act on various proteins, including proteins of the extracellular matrix (ECM) and connective tissue. In the normal wound healing process, proteases break down damaged ECM proteins and foreign material so that new tissue can form and wound closure can occur. Excessive wound proteases (e.g., matrix metalloproteinases (MMPs)) lead to the breakdown of newly formed ECM and other proteins, and as a result, wound healing is impaired due to damage to the ECM and abnormal prolongation of the inflammatory stage. In addition, proteases such as human neutrophil elastase (HNE) produced by neutrophils are responsible for fibronectin degradation in non-healing wounds. Furthermore, some bacteria that infect wounds secrete additional proteases. Thus, the activity levels of the proteases e.g., MMPs, HNE and bacterial proteases) can provide an indication on the stage of the wound: healing, inflammation and/or infection.

[0065] The present disclosure relates generally to wound dressing compositions that include at least one type of microcapsules comprising biocompatible markers and encapsulating materials that are capable of degradation by the wound proteases (e.g., MMPs, HNE and bacterial proteases), thereby releasing the biocompatible markers. Detection of the released biocompatible markers, along with results of color bleeding, results in a colorimetric scale that allows determination of type and activity levels of proteases active in the wound bed. For example, when the first microcapsules encapsulate a yellow dye, the second microcapsules encapsulate a blue dye, and third microcapsules encapsulate a red dye, and detection of yellow, red, green, orange, green, or purple color on the sealing layer may indicate activity of one or more type of proteases. In some embodiments, the intensity of the color may be proportional to the level of the protease activity. Intensity of the color may indicate level of the activity. Such diagnosis may be helpful for the selection of a patient for additional therapy, such as of change of dressing, administration of one or more antibiotics, hospitalization, or a combination thereof. Different microcapsules disclosed herein show differing levels of sensitivities to different levels and/or types of wound proteases, which, in turn allows for the diagnosis of state of inflammation, infection and/or healing of wound. [0066] For example, certain microcapsules disclosed herein may comprise an encapsulating material such as gelatin, which is an optimal substrate of the wound proteases. Other type of the microcapsules disclosed herein may comprise an encapsulating material such as collagen, which is degraded slower than gelatin. Yet other types of the microcapsules disclosed herein may comprise an encapsulating material comprising a component of the extracellular matrix, including, but not limited to one or more types of fibrillar collagens, elastin, fibronectin, laminin, nidogen, and gelatin, each of which are degraded by a different spectrum of wound proteases, degradation of which may be indicative of the state of inflammation, infection and/or healing of wound. Yet other types of the microcapsules disclosed herein may comprise an encapsulating material such as gelatin, in combination with oxidized regenerated cellulose, which provides therapeutic effect by decreasing pH of the wound. Any microcapsules disclosed herein may additionally comprise, without limitation, any beneficial components disclosed herein.

[0067] Each of the microcapsules disclosed herein may encapsulate different biocompatible markers, the release of which creates a colorimetric indicator of health of the wound. Nonlimiting examples of the biocompatible markers /differential indicator reagents include dyes (e.g., stains, pH indicators, redox indicators), pigments, fluorescent substances, enzyme substrates (e.g., chromogenic or fluorogenic substrates for proteases, or other enzymes found in wounds).

[0068] Accordingly, the wound dressing compositions of the present technology will advantageously allow for monitoring the levels of different proteases (e.g., MMPs, HNE and bacterial proteases) in a wound in a subject in need thereof, thereby providing an indication on the stage of the wound: healing, inflammation and/or infection. Accordingly, he wound dressing compositions of the present technology will advantageously facilitate selection of the patients suspected of, or diagnosed as having an elevated level of inflammatory mediators in a wound for treatment with a therapeutic regimen.

The First Microcapsules

[0069] The present disclosure provides a wound dressing composition that includes first microcapsules, comprising a first biocompatible marker encapsulated by a first encapsulating material comprising a first collagen.f

[0070] The first collagen may include a mammalian collagen, such as a bovine collagen, a human collagen, or a combination thereof. The first collagen of any embodiment herein may be a Type I collagen, a Type II collagen, a Type III collagen, may be obtained from any natural source, may be chemically-modified collagen (e.g., an atelocollagen obtained by removing the immunogenic telopeptides from natural collagen), or may be a combination of any two or more thereof. For example, the first collagen may include collagen obtained from bovine corium that has been rendered largely free of non-collagenous components, for example, including fat, non-collagenous proteins, polysaccharides, and other carbohydrates, such as by procedures described in U.S. Pat. Nos.

4,614,794 and 4,320,201, each of which is incorporated herein by reference. The bovine collagen may include one or both of bovine collagen type I and bovine collagen type III. In any embodiment disclosed herein, the first collagen may include a weight ratio of human collagen type I to human collagen type III of about 100:0, about 90: 10, about 80:20, about 70:30, about 60:40, about 50:50, about 40:60, about 30:70, about 20:80, about 10:90, about 0: 100, or any range including and/or in between any two of these values. The ratio by weight of human collagen type I to human collagen type III may be greater than about 50:50, or greater than about 70:30. The first collagen of any embodiment herein may include a weight ratio of type I bovine collagen to type III bovine collagen of about 95:5, about 85: 15, about 75:25, about 65:35, about 55:45, about 50:50, about 45:55, about 65:35, about 75:25, about 85:15, about 95:5, or any range including and/or in between any two of these values. In some embodiments, the ratio by weight of the type I bovine collagen to type III bovine collagen may be about 85: 15.

[0071] The first microcapsules may encapsulate a first biocompatible marker. Nonlimiting examples of first biocompatible markers present in the first microcapsules include dyes, stains, pigments, fluorescent substances, pH indicators, redox indicators, enzyme substrates (e.g., chromogenic or fluorogenic substrates for proteases, or other enzymes found in wounds). In some embodiments, first biocompatible markers present in the first microcapsules are soluble in water, saline or wound exudate. Nonlimiting examples of first biocompatible markers present in the first microcapsules are carotenoids (E160, E161, E164), chlorophyllin (E140, E141), anthocyanins (E163), and betanin (El 62), FD&C Blue #1, FD&C Blue # 2, FD&C Red # 40, FD&C Red #3, D&C Red #33, FD&C Yellow #5, FD&C Yellow #6, D&C Yellow #8, D&C Yellow #10, FD&C Green # 3, D&C Green #5, D&C Green # 8, and D&C External Violet # 2. Nonlimiting examples of biocompatible pH indicator dyes present in the first microcapsules as first biocompatible marker are anthocyanins, 2-fluoro-4-[4-(2-hydroxyethanesulfonyl)-phenylazo]-6-methoxyphenol (GJM-492) bromcresol green, and bromocresol purple, chlorphenol red, bromthymol blue, phenyl red, and thymol blue. Nonlimiting examples of fluorescent dyes present in the first microcapsules as first biocompatible marker are allura red, sunset yellow, brilliant blue, fast green, tartrazine, FD&C Red #3, D&C Red #22, D&C Red #28, D&C Yellow #8, D&C Orange #5, D&C Orange #11 and D&C Green #8.

[0072] The first microcapsules may additionally comprise, without limitation, any beneficial components disclosed herein.

[0073] The first microcapsules are designed to release the first biocompatible marker and/or the beneficial components when the first collagen is degraded by the proteases present in the wound bed (e.g., MMPs, HNE and bacterial proteases). The first biocompatible marker may be released when about 5 to about 50% of the first collagen is degraded. In some embodiments, the first microcapsules are designed to release detectable levels of the first biocompatible marker upon degradation of about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, or any range including and/or in between any two of these values of collagen. Upon release, the first biocompatible marker is designed to be wicked by capillary action through the substrate layer and optionally the wicking layer to the environmentally facing side of the wicking layer and/or the substrate sheet.

[0074] In any embodiment disclosed herein, the first microcapsules may have a diameter from about 0.01 pm to about 1000 pm. Thus, the first microcapsules may have a diameter from about 0.01 pm, about 0.03 pm, about 0.1 pm, about 0.3 pm, about 1 pm, about 3 pm, about 10 pm, about 30 pm, about 100 pm, about 300 pm, about 1000 pm, or any range including and/or in between any two of these values.

[0075] In any embodiment disclosed herein, the wound dressing composition may include about 0.5 wt.% to about 50 wt.% the first microcapsules. Additionally or alternatively, in some embodiments, the wound dressing composition may include about 1 wt.% to about 10 wt.% first microcapsules. Thus, the wound dressing composition may include about 0.5 wt.%, about 0.6 wt.%, about 0.7 wt.%, about 0.8 wt.%, about 0.9 wt.%, about 1 wt.%, about 2 wt.%, about 3 wt.%, about 4 wt.%, about 5 wt.%, about 6 wt.%, about 7 wt.%, about 8 wt.%, about 9 wt.%, about 10 wt.%, about 11 wt.%, about 12 wt.%, about 13 wt.%, about 14 wt.%, about 15 wt.%, about 16 wt.%, about 17 wt.%, about 18 wt.%, about 19 wt.%, about 20 wt.%, about 22 wt.%, about 24 wt.%, about 26 wt.%, about 28 wt.%, about 30 wt.%, about 32 wt.%, about 34 wt.%, about 36 wt.%, about 38 wt.%, about 40 wt.%, about 42 wt.%, about 44 wt.%, about 46 wt.%, about 48 wt.%, about 50 wt.%, or any range including and/or in between any two of these values of the first microcapsules.

The Second Microcapsules

[0076] The present disclosure provides a wound dressing composition that includes second microcapsules, comprising a second encapsulating material comprising a gelatin and an ORC, and a third encapsulating material comprising a second collagen, wherein the third encapsulating material encapsulates the second encapsulating material.

[0077] Rigidity of a gelatin film made of gelatin is expressed as Bloom strength, which is measured using a Bloom Gelometer. Higher Bloom values indicate the higher melting and gelling points of a gel, and the shorter gelling times. The second encapsulating material comprises a gelatin having a Bloom strength in the range of 50-325. In some embodiments, the second encapsulating material comprises a low Bloom gelatin, having a Bloom strength in the range of 50-150 and an average molecular mass of 20,000 to 25, 000; a medium Bloom gelatin, having a Bloom strength in the range of 175-225 and an average molecular mass of 40,000 to 50, 000; a high Bloom gelatin, having a Bloom strength in the range of 225-325 and an average molecular mass of 50,000 to 100, 000; or a combination thereof.

[0078] The first encapsulating material of the second microcapsule may comprise 30 wt.% to about 95 wt.% of a gelatin, and about 30 wt.% to about 70 wt.% oxidized regenerated cellulose (ORC) with a weight-average molecular weight of about 50,000 to about 1,000,000. In any embodiment disclosed herein, second encapsulating material may include from about 30 wt.% to about 70 wt.% of ORC. Thus, ORC may be included in the second encapsulating material in an amount of about 30 wt.%, about 32 wt.%, about 34 wt.%, about 36 wt.%, about 38 wt.%, about 40 wt.%, about 42 wt.%, about 44 wt.%, about 46 wt.%, about 48 wt.%, about 50 wt.%, about 52 wt.%, about 54 wt.%, about 56 wt.%, about 58 wt.%, about 60 wt.%, about 62 wt.%, about 64 wt.%, about 66 wt.%, about 68 wt.%, about 70 wt.%, or any range including and/or in between any two of these values. Also, a gelatin may be included in the second encapsulating material in an amount of about 30 wt.%, about 32 wt.%, about 34 wt.%, about 36 wt.%, about 38 wt.%, about 40 wt.%, about 42 wt.%, about 44 wt.%, about 46 wt.%, about 48 wt.%, about 50 wt.%, about 52 wt.%, about 54 wt.%, about 56 wt.%, about 58 wt.%, about 60 wt.%, about 62 wt.%, about 64 wt.%, about 66 wt.%, about 68 wt.%, about 70 wt.%, about 72 wt.%, about 74 wt.%, about 76 wt.%, about 78 wt.%, about 80 wt.%, about 82 wt.%, about 84 wt.%, about 86 wt.%, about 88 wt.%, about 90 wt.%, about 92 wt.%, about 94 wt.%, about 95 wt.%, or any range including and/or in between any two of these values.

[0079] The third encapsulating material may comprise a second collagen, which may include a mammalian collagen, such as a bovine collagen, a human collagen, or a combination thereof. The second collagen of any embodiment herein may be a Type I collagen, a Type II collagen, a Type III collagen, may be obtained from any natural source, may be chemically-modified collagen (e.g., an atelocollagen obtained by removing the immunogenic telopeptides from natural collagen), or may be a combination of any two or more thereof. For example, the second collagen may include collagen obtained from bovine corium that has been rendered largely free of non-collagenous components, for example, including fat, non-collagenous proteins, polysaccharides, and other carbohydrates, such as by procedures described in U.S. Pat. Nos. 4,614,794 and 4,320,201, each of which is incorporated herein by reference. The bovine collagen may include one or both of bovine collagen type I and bovine collagen type III. In any embodiment disclosed herein, the second collagen may include a weight ratio of human collagen type I to human collagen type III of about 100:0, about 90: 10, about 80:20, about 70:30, about 60:40, about 50:50, about 40:60, about 30:70, about 20:80, about 10:90, about 0: 100, or any range including and/or in between any two of these values. The ratio by weight of human collagen type I to human collagen type III may be greater than about 50:50, or greater than about 70:30. The second collagen of any embodiment herein may include a weight ratio of type I bovine collagen to type III bovine collagen of about 95:5, about 85:15, about 75:25, about 65:35, about 55:45, about 50:50, about 45:55, about 65:35, about 75:25, about 85: 15, about 95:5, or any range including and/or in between any two of these values. The ratio by weight of the type I bovine collagen to type III bovine collagen may be about 85: 15.

[0080] The second microcapsules may encapsulate a second biocompatible marker. Nonlimiting examples of second biocompatible markers present in the second microcapsules include dyes, stains, pigments, fluorescent substances, pH indicators, redox indicators, enzyme substrates (e.g., chromogenic or fluorogenic substrates for proteases, or other enzymes found in wounds). In some embodiments, second biocompatible markers present in the second microcapsules are soluble in water, saline or wound exudate. Nonlimiting examples of second biocompatible markers present in the second microcapsules are carotenoids (El 60, E161, El 64), chlorophyllin (E 140, E 141), anthocyanins (El 63), and betanin (El 62) FD&C Blue #1, FD&C Blue # 2, FD&C Red # 40, FD&C Red #3, D&C Red #33, FD&C Yellow #5, FD&C Yellow #6, D&C Yellow #8, D&C Yellow #10, FD&C Green # 3, D&C Green #5, D&C Green # 8, and D&C External Violet # 2. Nonlimiting examples of biocompatible pH indicator dyes present in the second microcapsules as second biocompatible marker are anthocyanins, 2-fluoro-4-[4-(2-hydroxyethanesulfonyl)-phenylazo]-6- methoxyphenol (GJM-492) bromcresol green, and bromocresol purple, chlorphenol red, bromthymol blue, phenyl red, and thymol blue. Nonlimiting examples of fluorescent dyes present in the second microcapsules as second biocompatible marker are allura red, sunset yellow, brilliant blue, fast green, tartrazine, FD&C Red #3, D&C Red #22, D&C Red #28, D&C Yellow #8, D&C Orange #5, D&C Orange #11 and D&C Green #8.

[0081] The second microcapsules may additionally comprise, without limitation, any beneficial components disclosed herein.

[0082] The second microcapsules are designed to release the second biocompatible marker and/or the beneficial components when the gelatin is degraded by the proteases present in the wound bed (e.g., MMPs, HNE and bacterial proteases). The second biocompatible marker may be released when about 5 to about 50% of the gelatin is degraded. In some embodiments, the second microcapsules are designed to release detectable levels of the second biocompatible marker upon degradation of about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, or any range including and/or in between any two of these values of gelatin. Upon release, the second biocompatible marker is designed to be wicked by capillary action through the substrate layer and optionally the wicking layer to the environmentally facing side of the wicking layer and/or the substrate sheet, to be viewed.

[0083] Additionally, or alternatively, the second microcapsules are designed to release the second biocompatible marker and/or the beneficial components when the second collagen is degraded by the proteases present in the wound bed (e.g., MMPs, HNE and bacterial proteases). The second biocompatible marker may be released when about 5 to about 50% of the second collagen is degraded. In some embodiments, the second microcapsules are designed to release detectable levels of the second biocompatible marker upon degradation of about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, or any range including and/or in between any two of these values of second collagen. Upon release, the second biocompatible marker is designed to be wicked by capillary action through the substrate layer and optionally the wicking layer to the environmentally facing side of the wicking layer and/or the substrate sheet.

[0084] In any embodiment disclosed herein, the wound dressing composition may include a plurality of second microcapsules. In any embodiment disclosed herein, the second microcapsules may have a diameter from about 0.01 pm to about 1000 pm. In any embodiment disclosed herein, the second microcapsules may have a diameter from about 0.01 pm to about 1000 pm. Thus, the second microcapsules may have a diameter from about 0.01 pm, about 0.03 pm, about 0.1 pm, about 0.3 pm, about 1 pm, about 3 pm, about 10 pm, about 30 pm, about 100 pm, about 300 pm, about 1000 pm, or any range including and/or in between any two of these values.

[0085] In any embodiment disclosed herein, the wound dressing composition may include about 0.5 wt.% to about 50 wt.% the second microcapsules. Additionally or alternatively, in some embodiments, the wound dressing composition may include about 1 wt.% to about 10 wt.% second microcapsules. Thus, the wound dressing composition may include about 0.5 wt.%, about 0.6 wt.%, about 0.7 wt.%, about 0.8 wt.%, about 0.9 wt.%, about 1 wt.%, about 2 wt.%, about 3 wt.%, about 4 wt.%, about 5 wt.%, about 6 wt.%, about 7 wt.%, about 8 wt.%, about 9 wt.%, about 10 wt.%, about 11 wt.%, about 12 wt.%, about 13 wt.%, about 14 wt.%, about 15 wt.%, about 16 wt.%, about 17 wt.%, about 18 wt.%, about 19 wt.%, about 20 wt.%, about 22 wt.%, about 24 wt.%, about 26 wt.%, about 28 wt.%, about 30 wt.%, about 32 wt.%, about 34 wt.%, about 36 wt.%, about 38 wt.%, about 40 wt.%, about 42 wt.%, about 44 wt.%, about 46 wt.%, about 48 wt.%, about 50 wt.%, or any range including and/or in between any two of these values of the second microcapsules.

Additional Microcapsules

[0086] The present disclosure provides a wound dressing composition that includes optional additional microcapsules. The additional microcapsules may comprise an encapsulating material comprising a component of the extracellular matrix, including, but not limited to one or more types of fibrillar collagens, elastin, fibronectin, and laminin, degradation of which may be indicative of the state of inflammation, infection and/or healing of wound. The additional microcapsules may encapsulate an additional biocompatible marker. The nonlimiting examples of first biocompatible marker and/or second biocompatible marker are also nonlimiting examples of one or more additional biocompatible markers disclosed herein. The additional microcapsules may comprise, without limitation, any beneficial components disclosed herein.

[0087] The additional microcapsules are designed to release the additional biocompatible marker and/or the beneficial components when the encapsulating material of the additional microcapsules are degraded by the proteases present in the wound bed ( e.g . , MMPs, HNE and bacterial proteases). The additional biocompatible marker and/or the beneficial components may be released when about 5 to about 50% of the encapsulating material of the additional microcapsules are degraded. In some embodiments, the additional microcapsules are designed to release detectable levels of the additional biocompatible marker and/or the beneficial components upon degradation of about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, or any range including and/or in between any two of these values of the encapsulating material of the additional microcapsules. Upon release, the additional biocompatible markers are designed to be wicked by capillary action through the substrate layer and optionally the wicking layer to the environmentally facing side of the wicking layer and/or the substrate sheet. Beneficial Components

[0088] The microcapsules of any embodiment disclosed herein may comprise, without limitation, any beneficial components disclosed herein.

[0089] In any embodiments disclosed herein, the first microcapsules, the second microcapsules and/or the additional microcapsules may comprise a silver compound. In any embodiments disclosed herein, the first microcapsules, the second microcapsules and/or the additional microcapsules may comprise a silver compound. Additionally or alternatively, in some embodiments disclosed herein, the first microcapsules, the second microcapsules and/or the additional microcapsules may comprise about 0.1 wt.% to about 3 wt.% of a silver compound. Additionally or alternatively, in some embodiments disclosed herein, the first microcapsules, the second microcapsules and/or the additional microcapsules may comprise about 0.1 wt.%, about 0.25 wt.%, about 0.50 wt.%, about 0.75 wt.%, about 0.8 wt.%, about 0.85 wt.%, about 0.9 wt.%, about 0.95 wt.%, about 1 wt.%, about 1.25 wt.%, about 1.5 wt.%, about 1.75 wt.%, about 2 wt.%, about 2.25 wt.%, about 2.5 wt.%, about 2.75 wt.%, about 3 wt.%, or any range including and/or in between any two of the preceding values, of the silver compound.

[0090] In any embodiments disclosed herein, the silver compound of the first microcapsules, the second microcapsules and/or the additional microcapsules may comprise one or more pharmaceutically acceptable silver salts. Exemplary one or more pharmaceutically acceptable silver salts of the microcapsules include, but are not limited to, silver oxide, silver oxysalts, silver chromate, silver allantoinate, silver borate, silver glycerolate, silver nitrate, silver acetate, silver chloride, silver sulfate, silver lactate, silver bromide, silver iodide, silver carbonate, silver citrate, silver laurate, silver deoxycholate, silver salicylate, silver / aminobenzoate. silver /^-aminosalicylate. nanocrystalline silver, any pharmaceutically acceptable salt thereof, or any combination thereof. Additionally or alternatively, in some embodiments disclosed herein, the silver compound of the first microcapsules, the second microcapsules and/or the additional microcapsules comprises a silver (II) oxide, silver (III) oxide, a silver oxy-salt, or any combination thereof. Additionally or alternatively, in some embodiments disclosed herein, the silver oxy-salt may comprise a general formula of Ag(Ag₃0_a)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.

[0091] In any embodiment of the microcapsules disclosed herein, the first microcapsules, the second microcapsules and/or the additional microcapsules may comprise about 1 wt.% to about 15 wt.% of at least one plasticizer. Additionally or alternatively, in some embodiments disclosed herein, the first microcapsules, the second microcapsules and/or the additional microcapsules may comprise about 1 wt.%, about 1.5 wt.%, about 2 wt.%, about 2.5 wt.%, about 3 wt.%, about 3.5 wt.%, about 4 wt.%, about 4.5 wt.%, about 5 wt.%, about 5.5 wt.%, about 6 wt.%, about 6.5 wt.%, about 7 wt.%, about 7.5 wt.%, about 8 wt.%, about 8.5 wt.%, about 9 wt.%, about 9.5 wt.%, about 10 wt.%, about 10.5 wt.%, about 11 wt.%, about 11.5 wt.%, about 12 wt.%, about 12.5 wt.%, about 13 wt.%, about 13.5 wt.%, about 14 wt.%, about 14.5 wt.%, about 15 wt.%, or any range including and/or in between any two of the preceding values, of the at least one plasticizer. Exemplary plasticizers include, but are not limited to, an acetylated monoglyceride, an alkyl citrate, methyl ricinoleate, glycerol, polyvinylpyrrolidone, and any combination thereof. Examples of alkyl citrates include, but are not limited to, 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.

[0092] Additionally, or alternatively, in some embodiments, the first microcapsules, the second microcapsules and/or the additional microcapsules comprise a silver compound. In some embodiments, the first microcapsules, the second microcapsules and/or the additional microcapsules comprise about 0.1 wt.% to about 3 wt.% of the silver compound. In some embodiments, the silver compound comprises one or more pharmaceutically acceptable silver salts. In some embodiments, the one or more pharmaceutically acceptable silver salts is selected from the group consisting of silver oxide, silver oxysalts, silver chromate, silver allantoinate, silver borate, silver glycerolate, silver nitrate, silver acetate, silver chloride, silver sulfate, silver lactate, silver bromide, silver iodide, silver carbonate, silver citrate, silver laurate, silver deoxycholate, silver salicylate, silver p-aminobenzoate, silver p-aminosalicylate, nanocrystalline silver, any pharmaceutically acceptable salt thereof, and any combination thereof.

[0093] Additionally, or alternatively, in some embodiments, the microcapsules further comprise one or more additional additives selected from the group consisting of an antimicrobial agent, an antioxidant, a signaling protein, and a combination any two or more thereof.

[0094] Additionally, or alternatively, in some embodiments, the first microcapsules, the second microcapsules and/or the additional microcapsules comprise about 0.001 wt.% to about 5 wt.% of the antimicrobial agent. In some embodiments, the antimicrobial agent comprises one or more of tetracycline, penicillins, terramycins, erythromycin, bacitracin, neomycin, polymycin B, mupirocin, clindamycin, colloidal silver, silver sulfadiazine, chlorhexidine, povidone iodine, triclosan, sucralfate, quaternary ammonium salts, pharmaceutically acceptable silver salts, or any combination thereof. [0095] Additionally, or alternatively, in some embodiments, the first microcapsules, the second microcapsules and/or the additional microcapsules comprise about 0.001 wt.% to about 5 wt.% of the antioxidant. In some embodiments, the antioxidant comprises one or more of anthocyanins, astaxanthin, bilirubin, canthaxanthin, capsaicin, citric acid, curcumin, coenzyme Q10, eugenol, flavanols, flavonolignans, flavanones, flavones, flavonols, iodide, isoflavone phytoestrogens, lutein, lycopene, manganese, melatonin, N-acetylcysteine, oxalic acid, phenolic acids, phytic acid, R-a-lipoic acid, stilbenoids, tocopherol, tocotrienol, vitamin A, vitamin C, vitamin E, xanthones, zeaxanthin, a- carotene, b-carotene, or any combination thereof. In some embodiments, the anthocyanins are selected from the group consisting of cyanidin, delphinidin, malvidin, pelargonidin, peonidin, petunidin, and any combination thereof. In some embodiments, the flavanols are selected from the group consisting of catechin, epicatechin, theaflavin, thearubigins, gallocatechin, epigallocatechin, or any gallate ester thereof, and any combination thereof. In some embodiments, the flavanones are selected from the group consisting of eriodictyol, hesperetin, naringenin, and any combination thereof. In some embodiment, the flavones are selected from the group consisting of apigenin, luteolin, tangeritin, and any combination thereof. In some embodiments, the flavonols are selected from the group consisting of isorhamnetin, kaempferol, myricetin, proanthocyanidins, quercetin, rutin, and any combination thereof. In some embodiments, the isoflavone phytoestrogens are selected from the group consisting of daidzein, genistein, glycitein, and any combination thereof. In some embodiments, 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. In some embodiments, the stilbenoids are selected from the group consisting of resveratrol, pterostilbene, and any combination thereof.

[0096] Additionally, or alternatively, in some embodiments, the first microcapsules, the second microcapsules and/or the additional microcapsules comprise about 0.001 wt.% to about 5 wt.% of the signaling protein. In some embodiments, the signaling protein comprises one or more of platelet- derived growth factor (PDGF), transforming growth factor beta (TGF ), fibroblast growth factors (FGFs), epidermal growth factor (EGF), or any combination thereof. In some embodiments, the fibroblast growth factors (FGFs) 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.

The Substrate Sheet

[0097] The present disclosure provides a wound dressing that includes a substrate sheet. Additionally, or alternatively, in some embodiments, the first microcapsules, the second microcapsules and/or the additional microcapsules are embedded in the substrate sheet. In some embodiments, the first microcapsules, the second microcapsules, the optional additional microcapsules are embedded in a portion of the substrate sheet. Additionally, or alternatively, the first microcapsules, the second microcapsules and/or the additional microcapsules are coupled to the substrate sheet. In some embodiments, the first microcapsules, the second microcapsules, the optional additional microcapsules are coupled to the wound facing side of the substrate sheet. In some embodiments, the first microcapsules, the second microcapsules, the optional additional microcapsules are coupled to the entire wound facing side of the substrate sheet. In some embodiments, the first microcapsules, the second microcapsules, the optional additional microcapsules are coupled to a portion of the wound facing side of the substrate sheet.

[0098] Additionally, or alternatively, the substrate sheet comprises a plurality of perforations to allow passage of fluids. The perforations create holes in the substrate sheet. In some embodiments, the perforations are laser cut. The perforations may have any two dimensional shape, such as circular, triangular, or irregularly shaped. In some embodiments, the perforations are circular in shape.

[0099] The substrate sheet is designed to contact the wound, and thereby promote the contact of wound with the microcapsules. The substrate sheet is also designed to transport wound exudate to the wick layer via capillary action. The substrate sheet is also designed to promote transport of the biocompatible markers released upon degradation of the microcapsules through the substrate layer, optionally to the wicking layer via capillary action. In any embodiment of the wound dressing composition disclosed herein, the substrate sheet may include a wound-facing side and an environmental -facing side.

[00100] The substrate sheet comprises a dressing material. Any dressing material known in the art is suitable for the substrate sheet. Nonlimiting examples of suitable dressing material include a polymer film (e.g., polyurethane film), a textile, such as a medical textile (e.g., Asahi nylon), and cellulose.

[00101] Additionally or alternatively, in some embodiments, the substrate sheet may be any of the layers conventionally used to form layers over a wound contacting layer in a laminated wound dressing, for example absorbent layer or backing layer. In certain embodiments, the substrate sheet is a backing layer in the form of a sheet of continuous semipermeable or impermeable polymer. In some embodiments, the substrate sheet may be an absorbent layer for example a hydrophilic foam, a sponge, a film, or a textile layer. The textile may be nonwoven, knitted or woven. In some embodiments, the substrate sheet is (a) a semipermeable or impermeable polymer film, or (b) a hydrophilic foam sheet, or (c) a nonwoven web.

[00102] Suitable semipermeable or impermeable polymer films for the substrate sheet include any of the semipermeable films conventionally used to form a backing sheet of wound dressings. The films may be continuous, i.e. they do not comprise macroscopic apertures that would allow passage of wound fluid. In some embodiments, continuous conformable substrate sheets of this type will suitably have a moisture vapor transmission rate (MVTR) of the substrate sheet alone of 300 to 35000 g/m²/24hrs, suitably 500 to 25000 g/m²/24hrs at 37.5 °C at 100% to 10% relative humidity difference (measured before coating with the silicone adhesive). Additionally or alternatively, in some embodiments, the substrate sheet may be microorganism-impermeable.

[00103] Nonlimiting examples of suitable polymers for forming the substrate sheet include polyurethanes and poly alkoxyalkyl acrylates and methacrylates. In some embodiments, the substrate sheet comprises a continuous layer of a high density blocked polyurethane foam that is predominantly closed-cell. A nonlimiting example of suitable backing sheet material is the polyurethane film available under the Registered Trade Mark ESTANE™ 5714F. Also suitable are elastomeric polymeric esters such as Du Pont HYTREL™ (Registered Trade Mark).

[00104] Suitable hydrophilic foam sheets for use as the substrate sheet include polyurethane foams, carboxylated butadiene-styrene rubber, polyacrylate, polyvinylic or cellulosic foams. The hydrophilic foam may be open-cell or closed-cell. In some embodiments, the foam comprises a polyurethane, and more suitably it comprises at least 50% by weight of one or more polyurethanes, for example at least 75% by weight thereof.

[00105] The hydrophilic foams used in the substrate sheets may also have the property of swelling and expanding when water is absorbed. The degree of swelling of the hydrophilic foams on complete saturation with an aqueous medium is typically at least 100% (expressed in terms of increase in volume), and suitably at least 200%. In some embodiments, the foams swell by 400 to 800%.

Despite this high degree of swelling, however, the foams retain their integrity even after absorption of large quantities of water. In some embodiments, the cells of the hydrophilic foams have an average diameter in the range 0.1 to 0.6 mm. Some suitable hydrophilic foams are as described in EP-A- 0541391. These foam layers are available from Systagenix Wound Management under the Registered Trade Marks TIELLE and HYPOL.

[00106] In some embodiments, the basis weight of the hydrophilic foam when used as a substrate sheet in the materials of the present technology is from 0.2 to 1.5 kg/m², more suitably 0.5 to 1.0 kg/m².

[00107] Suitable textiles for use as the substrate sheet include any of those conventionally used for absorbent products, including cellulose woven or nonwoven webs, or cellulose derivatives such as viscose, rayon or oxidized regenerated cellulose. In certain embodiments, the fabric comprises at least about 10wt.% of hydrogel-forming absorbent fibers based on the dry weight of the fabric, for example, the fabric comprises at least about 20wt.% of the hydrogel-forming fibers, for example from about 30wt.% to about 50wt.% of such fibers.

[00108] The term “hydrogel-forming fibers” refers to fibers that can absorb at least about twice their own weight of water, suitably at least about four times their own weight of water, to form a hydrogel. The fibers are normally insoluble in water. Non-limiting examples of suitable materials for the hydrogel-forming fibers include alginates, carboxymethylcelluloses, hydroxyethyl-celluloses, carboxymethylcellulose, polyacrylates, and hyaluronates.

[00109] The size, shape and thickness of the substrate sheet may be varied as desired. It is recognized that a thinner substrate sheet will allow detection of low levels of wound proteases, and thicker substrate sheets will be suitable to detect high levels of wound proteases. In any embodiment of the wound dressing composition disclosed herein, the substrate sheet may have a thickness of about 0.5 mm to about 30 mm. Additionally or alternatively, in some embodiments of the wound dressing composition disclosed herein, the substrate sheet may have a thickness of about 0.5 mm, about 1 mm, about 1.5 mm, about 2 mm, about 2.5 mm, about 3 mm, about 5 mm, about 7.5 mm, about 10 mm, about 15 mm, about 20 mm, about 25 mm, about 30 mm, or any range including and/or in between any two of the preceding values. In any embodiment of the wound dressing composition disclosed herein, the substrate sheet may have a thickness of about 0.5 mm to about 2 mm, about 1 mm to about 4 mm, about 2 mm to about 6 mm, about 2.5 mm to about 10 mm, 5 mm to about 20 mm, about 10 mm to about 30 mm, or any range including and/or in between any two of these values.

[00110] In any embodiment of the wound dressing composition disclosed herein, the biopolymer of the substrate sheet comprise a collagen, an oxidized cellulose, a polysaccharide, chitosan, gelatin, hyaluronic acid, or a combination of any two or more thereof.

[00111] In any embodiment of the wound dressing composition disclosed herein, the substrate sheet may comprise about 0.1 wt.% to about 100 wt.% a biopolymer. Additionally or alternatively, in some embodiments of the wound dressing disclosed herein, the amount of biopolymer in the substrate sheet may be about 0.1 wt.%, about 0.5 wt.%, about 1 wt.%, about 2.5 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.%, about 98 wt. %, about 99 wt. %, about 100 wt.%, or any range including and/or in between any two of the preceding values.

[00112] In any embodiment of the wound dressing composition disclosed herein, the solid content of the substrate sheet may comprise about 0.1 wt.% to about 10 wt.%. Additionally or alternatively, in some embodiments of the wound dressing disclosed herein, the solid content of the substrate sheet may comprise about 0.1 wt.%, about 0.5 wt.%, about 1 wt.%, about 1.5 wt.%, about 2 wt.%, about 2.5 wt.%, about 3 wt.%, about 3.5 wt.%, about 4 wt.%, about 4.5 wt.%, about 5 wt.%, about 5.5 wt.%, about 6 wt.%, about 6.5 wt.%, about 7 wt.%, about 7.5 wt.%, about 8 wt.%, about 8.5 wt.%, about 9 wt.%, about 9.5 wt.%, about 10 wt.%, or any range including and/or in between any two of the preceding values.

[00113] In any embodiment of the wound dressing composition disclosed herein, the substrate sheet may comprise a silver compound. Additionally or alternatively, in some embodiments of the wound dressing disclosed herein, the substrate sheet may comprise about 20 wt.% to about 80 wt.% of a collagen. Additionally or alternatively, in some embodiments of the wound dressing disclosed herein, the substrate sheet may comprise 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.%, or any range including and/or in between any two of the preceding values, of the collagen. Additionally or alternatively, in some embodiments of the wound dressing disclosed herein, the substrate sheet may comprise about 20 wt.% to about 40 wt.%, about 30 wt.% to about 50 wt.%, about 40 wt.% to about 60 wt.%, about 50 wt.% to about 70 wt.%, about 60 wt.% to about 80 wt.% of the collagen. [00114] In any embodiment of the wound dressing composition disclosed herein, the substrate sheet may comprise a silver compound. Additionally or alternatively, in some embodiments of the wound dressing disclosed herein, the substrate sheet may comprise about 20 wt.% to about 80 wt.% of an oxidized cellulose. Additionally or alternatively, in some embodiments of the wound dressing disclosed herein, the substrate sheet may comprise 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.%, or any range including and/or in between any two of the preceding values, of the oxidized cellulose. Additionally or alternatively, in some embodiments of the wound dressing disclosed herein, the substrate sheet may comprise about 20 wt.% to about 40 wt.%, about 30 wt.% to about 50 wt.%, about 40 wt.% to about 60 wt.%, about 50 wt.% to about 70 wt.%, about 60 wt.% to about 80 wt.% of the oxidized cellulose.

[00115] In any embodiment of the wound dressing composition disclosed herein, the substrate sheet may comprise a silver compound. Additionally or alternatively, in some embodiments of the wound dressing disclosed herein, the substrate sheet may comprise about 0.1 wt.% to about 3 wt.% of a silver compound. Additionally or alternatively, in some embodiments of the wound dressing disclosed herein, the substrate sheet may comprise about 0.1 wt.%, about 0.25 wt.%, about 0.50 wt.%, about 0.75 wt.%, about 0.8 wt.%, about 0.85 wt.%, about 0.9 wt.%, about 0.95 wt.%, about 1 wt.%, about 1.25 wt.%, about 1.5 wt.%, about 1.75 wt.%, about 2 wt.%, about 2.25 wt.%, about 2.5 wt.%, about 2.75 wt.%, about 3 wt.%, or any range including and/or in between any two of the preceding values, of the silver compound.

[00116] In any embodiment of the wound dressing composition disclosed herein, the silver compound of the substrate sheet comprises one or more pharmaceutically acceptable silver salts. Exemplary sources of the one or more pharmaceutically acceptable silver salts of the substrate sheet include, but are not limited to, silver oxide, silver oxysalts, silver chromate, silver allantoinate, silver borate, silver glycerolate, silver nitrate, silver acetate, silver chloride, silver sulfate, silver lactate, silver bromide, silver iodide, silver carbonate, silver citrate, silver laurate, silver deoxycholate, silver salicylate, silver / aminobenzoate. silver /^-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 of the substrate sheet 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(Ag₃0_a)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.

[00117] In any embodiment of the wound dressing composition disclosed herein, the substrate sheet may comprise about 1 wt.% to about 15 wt.% of at least one plasticizer. Additionally or alternatively, in some embodiments of the wound dressing disclosed herein, the substrate sheet may comprise about 1 wt.%, about 1.5 wt.%, about 2 wt.%, about 2.5 wt.%, about 3 wt.%, about 3.5 wt.%, about 4 wt.%, about 4.5 wt.%, about 5 wt.%, about 5.5 wt.%, about 6 wt.%, about 6.5 wt.%, about 7 wt.%, about 7.5 wt.%, about 8 wt.%, about 8.5 wt.%, about 9 wt.%, about 9.5 wt.%, about 10 wt.%, about 10.5 wt.%, about 11 wt.%, about 11.5 wt.%, about 12 wt.%, about 12.5 wt.%, about 13 wt.%, about 13.5 wt.%, about 14 wt.%, about 14.5 wt.%, about 15 wt.%, or any range including and/or in between any two of the preceding values, of the at least one plasticizer. Exemplary plasticizers include, but are not limited to, an acetylated monoglyceride, an alkyl citrate, methyl ricinoleate, glycerol, polyvinylpyrrolidone, and any combination thereof. Examples of alkyl citrates include, but are not limited to, 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.

[00118] In any embodiment of the dressing composition disclosed herein, the substrate sheet may comprise about 1 wt.% to about 15 wt.% of at least one plasticizer. Additionally or alternatively, in some embodiments disclosed herein, the microcapsules may comprise about 1 wt.%, about 1.5 wt.%, about 2 wt.%, about 2.5 wt.%, about 3 wt.%, about 3.5 wt.%, about 4 wt.%, about 4.5 wt.%, about 5 wt.%, about 5.5 wt.%, about 6 wt.%, about 6.5 wt.%, about 7 wt.%, about 7.5 wt.%, about 8 wt.%, about 8.5 wt.%, about 9 wt.%, about 9.5 wt.%, about 10 wt.%, about 10.5 wt.%, about 11 wt.%, about

11.5 wt.%, about 12 wt.%, about 12.5 wt.%, about 13 wt.%, about 13.5 wt.%, about 14 wt.%, about

14.5 wt.%, about 15 wt.%, or any range including and/or in between any two of the preceding values, of the at least one plasticizer. Exemplary plasticizers include, but are not limited to, an acetylated monoglyceride, an alkyl citrate, methyl ricinoleate, glycerol, polyvinylpyrrolidone, and any combination thereof. Examples of alkyl citrates include, but are not limited to, 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. [00119] Additionally, or alternatively, in some embodiments, the substrate sheet comprises a silver compound. In some embodiments, the substrate sheet comprises about 0.1 wt.% to about 3 wt.% of the silver compound. In some embodiments, the silver compound comprises one or more pharmaceutically acceptable silver salts. In some embodiments, the one or more pharmaceutically acceptable silver salts is selected from the group consisting of silver oxide, silver oxysalts, silver chromate, silver allantoinate, silver borate, silver glycerolate, silver nitrate, silver acetate, silver chloride, silver sulfate, silver lactate, silver bromide, silver iodide, silver carbonate, silver citrate, silver laurate, silver deoxycholate, silver salicylate, silver p-aminobenzoate, silver p-aminosalicylate, nanocrystalline silver, any pharmaceutically acceptable salt thereof, and any combination thereof. [00120] Additionally, or alternatively, in some embodiments, the substrate layer further comprises one or more additional additives selected from the group consisting of an antimicrobial agent, an antioxidant, a signaling protein, and a combination any two or more thereof. [00121] Additionally, or alternatively, in some embodiments, the substrate sheet comprises about 0.001 wt.% to about 5 wt.% of the antimicrobial agent. In some embodiments, the antimicrobial agent comprises one or more of tetracycline, penicillins, terramycins, erythromycin, bacitracin, neomycin, polymycin B, mupirocin, clindamycin, colloidal silver, silver sulfadiazine, chlorhexidine, povidone iodine, triclosan, sucralfate, quaternary ammonium salts, pharmaceutically acceptable silver salts, or any combination thereof.

[00122] Additionally, or alternatively, in some embodiments, the substrate sheet comprises about 0.001 wt.% to about 5 wt.% of the antioxidant. In some embodiments, the antioxidant comprises one or more of anthocyanins, astaxanthin, bilirubin, canthaxanthin, capsaicin, citric acid, curcumin, coenzyme Q10, eugenol, flavanols, flavonolignans, flavanones, flavones, flavonols, iodide, isoflavone phytoestrogens, lutein, lycopene, manganese, melatonin, N-acetylcysteine, oxalic acid, phenolic acids, phytic acid, R-a-lipoic acid, stilbenoids, tocopherol, tocotrienol, vitamin A, vitamin C, vitamin E, xanthones, zeaxanthin, a-carotene, b-carotene, or any combination thereof. In some embodiments, the anthocyanins are selected from the group consisting of cyanidin, delphinidin, malvidin, pelargonidin, peonidin, petunidin, and any combination thereof. In some embodiments, the flavanols are selected from the group consisting of catechin, epicatechin, theaflavin, thearubigins, gallocatechin, epigallocatechin, or any gallate ester thereof, and any combination thereof. In some embodiments, the flavanones are selected from the group consisting of eriodictyol, hesperetin, naringenin, and any combination thereof. In some embodiment, the flavones are selected from the group consisting of apigenin, luteolin, tangeritin, and any combination thereof. In some embodiments, the flavonols are selected from the group consisting of isorhamnetin, kaempferol, myricetin, proanthocyanidins, quercetin, rutin, and any combination thereof. In some embodiments, the isoflavone phytoestrogens are selected from the group consisting of daidzein, genistein, glycitein, and any combination thereof.

[00123] Additionally, or alternatively, in some embodiments, the substrate sheet comprises about 0.001 wt.% to about 5 wt.% of the signaling protein. In some embodiments, the signaling protein comprises one or more of platelet-derived growth factor (PDGF), transforming growth factor beta (TGF ), fibroblast growth factors (FGFs), epidermal growth factor (EGF), or any combination thereof. In some embodiments, the fibroblast growth factors (FGFs) 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.

The Wicking Layer

[00124] The present disclosure provides a wound dressing composition that includes an optional wicking layer. The wicking layer may include a wound-facing side and an environmental -facing side. In any embodiment disclosed herein, the environmental-facing side of the substrate layer is coupled to the wound-facing side of the wicking layer. The substrate sheet and the wicking layer are arranged such that when the substrate contacts fluid, capillary action draws the biocompatible markers to the wicking layer, which then transports the biocompatible markers to the sealing layer via capillary action. The wicking layer is designed to promote transport of the biocompatible markers, which are transported from the substrate layer to the wicking layer, to the sealing layer via capillary action. Accordingly, the wicking layer is configured to allow capillary action. The wicking layer may include any suitable liquid permeable material capable of absorbing fluids and wicking it to the sealing layer. In some embodiments, the wicking layer and/or the substrate sheet are not transparent, so that the biocompatible markers are not visible without being wicked to the sealing layer.

[00125] In any embodiment disclosed herein, the wicking layer is configured to wick wound exudate and other fluids from the wound bed towards a sealing layer, and to maintain the location of fluids within the sealing layer. Accordingly, the wicking layer is manufactured from one or more of an absorptive textile or paper, with a thickness and absorptivity tuned (e.g. by tailoring the amount of capillary action) such that a detection (for example, visual detection) of the biocompatible marker in the sealing layer corresponds to a predetermined amount of the activity of proteases present in the wound bed (e.g., MMPs, FINE and bacterial proteases). In some embodiments, the wicking layer comprises a thin layer of a non-woven material. Non-limiting examples of suitable non-woven material are 100% polyethylene or polyethylene terephthalate, such as Sawabond® 4383/4157. In some embodiments, the wicking layer comprises a high wicking material, such as Milliken Interdry or Libeltex TDL2.

[00126] The size, shape and thickness of the wicking layer may be varied as desired. In any embodiment disclosed herein, the wicking layer may have a thickness of about 0.02 mm to about 2 mm. Additionally or alternatively, in some embodiments, the wicking layer may have a thickness of about 0.02 mm, about 0.04 mm, about 0.06 mm, about 0.08 mm, about 0.1 mm, about 0.15 mm, about 0.2 mm, about 0.3 mm, about 0.4 mm, about 0.5 mm, about 0.75 mm, about 1 mm, about 1.25 mm, about 1.5 mm, about 2 mm, or any range including and/or in between any two of the preceding values.

The Sealing Laver [00127] The present disclosure provides a wound dressing composition that includes a sealing layer. In some embodiments, the sealing layer includes a window for viewing the wicking layer and/or the substrate sheet. In some embodiments, the window is suitable for viewing first biocompatible marker and/or second biocompatible marker transported to the environmentally facing side of the substrate sheet and/or environmentally facing side of the wicking layer.

[00128] The sealing layer may include a wound-facing side and an environmental-facing side, and wherein the sealing layer may include a backing film, or a drape. In some embodiments, the drape comprises an arctic adhesive. In any embodiment disclosed herein, the environmental-facing side of the wicking layer and/or the substrate sheet is coupled to the wound-facing side of the sealing layer. [00129] The size, shape and thickness of the sealing layer may be varied as desired. In any embodiment disclosed herein, the sealing layer may have a thickness of about 10 pm to about 2000 pm. Additionally or alternatively, in some embodiments, the sealing layer may have a thickness of about 10 pm, about 25 pm, about 50 pm, about 75 pm, about 100 pm, about 105 pm. about 150 pm, about 200 pm, about 300 pm, about 400 pm, about 500 pm, about 750 pm, about 1000 pm, about 1250 pm, about 1500 pm, about 1750 pm, about 2000 pm, or any range including and/or in between any two of the preceding values.

[00130] In any embodiment disclosed herein, the sealing layer may be selected from the group consisting of polyurethane, polyalkoxy alkyl acrylate, polyalkoxy alkyl methacrylates, and any combination thereof.

[00131] In any embodiment disclosed herein, the sealing layer is substantially impermeable to liquid or wound exudate. Additionally or alternatively, in some embodiments, the sealing layer is microorganism impermeable. Additionally or alternatively, in some embodiments, the sealing layer is semi-permeable to water vapor.

[00132] In any embodiment disclosed herein, the sealing layer may comprise a moisture vapor transmission rate (MVTR) of about 450 g/m²/24hrs to about 20,000 g/m²/24hrs at 37.5°C at 50% relative humidity difference as described in ASTM E96-00. Additionally or alternatively, in some embodiments, the sealing layer may comprise a MVTR of about 450 g/m²/24hrs, about 500 g/m²/24hrs, about 600 g/m²/24hrs, about 750 g/m²/24hrs, about 1000 g/m²/24hrs, about 1250 g/m²/24hrs, about 1500 g/m²/24hrs, about 1750 g/m²/24hrs, about 2000 g/m²/24hrs, about 3000 g/m²/24hrs, about 3500 g/m²/24hrs, about 4000 g/m²/24hrs, about 5000 g/m²/24hrs, about 7500 g/m²/24hrs, about 10000 g/m²/24hrs, about 12500 g/m²/24hrs, about 15000 g/m²/24hrs, about 17500 g/m²/24hrs, about 20000 g/m²/24hrs, or any range including and/or in between any two of the preceding values. Such moisture vapor transmission rates allow the wound under the wound dressing to heal under moist conditions without causing the skin surrounding the wound to macerate.

The Wound Dressing

[00133] The wound dressing compositions of the present technology will advantageously allow for monitoring the levels of different proteases (e.g., MMPs, HNE and bacterial proteases) in a wound in a subject in need thereof, and thereby provide an indication on the stage of the wound: healing, inflammation and/or infection. The wound dressing compositions of the present technology are designed to promote contact of the microcapsules disclosed herein with the wound bed such that the proteases present in the wound bed (e.g., MMPs, HNE and bacterial proteases) will degrade the encapsulating materials of the microcapsules to differing degrees. For example, certain microcapsules disclosed herein may comprise an encapsulating material such as gelatin, which is an optimal substrate of the wound proteases. Other type of the microcapsules disclosed herein may comprise an encapsulating material such as collagen, which is degraded slower than gelatin. Yet other types of the microcapsules disclosed herein may comprise an encapsulating material comprising a component of the extracellular matrix, including, but not limited to one or more types of fibrillar collagens, elastin, fibronectin, laminin, nidogen, and gelatin, degradation of which may be indicative of the state of inflammation, infection and/or healing of wound. Yet other types of the microcapsules disclosed herein may comprise an encapsulating material such as gelatin, in combination with oxidized regenerated cellulose.

[00134] When the biocompatible markers present in different type of microcapsules are released, the substrate sheet and the wicking layer draw the biocompatible markers to the sealing layer via capillary action. Detection of the biocompatible markers on the substrate sheet and/or the wicking layer through the optional window allows the estimation of identity and range of activity of proteases present in the wound bed (e.g., MMPs, HNE and bacterial proteases). For example, in some cases digestion of differing levels of different types of microcapsules may provide a colorimetric visualization that shows a range of activity of proteases present in the wound bed (e.g., MMPs, HNE and bacterial proteases) and may provide a threshold indication for selection of the patient for additional therapy, such as of change of dressing, administration of one or more antibiotics, hospitalization, or a combination thereof.

[00135] In one aspect, the present disclosure provides a wound dressing composition comprising a substrate sheet; and a plurality of first microcapsules comprising a first biocompatible marker encapsulated by a first encapsulating material, wherein the first encapsulating material comprises a first collagen. Additionally or alternatively, in some embodiments, the wound dressing composition further comprises a wicking layer. Additionally or alternatively, in some embodiments, the wound dressing composition further comprises a sealing layer comprising a window for viewing the wicking layer and/or the substrate sheet.

[00136] In one aspect, the present disclosure provides a wound dressing composition comprising a substrate sheet; and a plurality of second microcapsules comprising a second biocompatible marker encapsulated by a second encapsulating material, wherein the second encapsulating material comprises a gelatin and an oxidized regenerated cellulose (ORC), and a third encapsulating material, wherein the third encapsulating material comprises a second collagen, wherein the third encapsulating material encapsulates the second encapsulating material. Additionally or alternatively, in some embodiments, the wound dressing composition further comprises a wicking layer. Additionally or alternatively, in some embodiments, the wound dressing composition further comprises a sealing layer comprising a window for viewing the wicking layer and/or the substrate sheet.

[00137] In one aspect, the wound dressing of the present technology comprises a substrate sheet; a plurality of first microcapsules comprising a first biocompatible marker encapsulated by a first encapsulating material, wherein the first encapsulating material comprises a first collagen; and a plurality of second microcapsules comprising a second biocompatible marker encapsulated by a second encapsulating material, wherein the second encapsulating material comprises a gelatin and an ORC, and a third encapsulating material, wherein the third encapsulating material comprises a second collagen, wherein the third encapsulating material encapsulates the second encapsulating material; optionally, a wicking layer; and a sealing layer comprising a window for viewing the wicking layer and/or the substrate sheet.

[00138] As noted above, the present disclosure provides a wound dressing composition that includes first microcapsules, second microcapsules, optional additional microcapsules, a substrate sheet, an optional wicking layer, and a sealing layer, wherein each of the substrate sheet, the optional wicking layer, and the sealing layer may include a wound-facing side and an environmental -facing side. [00139] In some embodiments, the first microcapsules, the second microcapsules, the optional additional microcapsules are embedded in the substrate sheet. Additionally, or alternatively, in some embodiments, the first microcapsules, the second microcapsules, the optional additional microcapsules are coupled to the substrate sheet. Additionally, or alternatively, in some embodiments, the first microcapsules, the second microcapsules, the optional additional microcapsules are coupled to the wound facing side of the substrate sheet.

[00140] In some embodiments, the wound-facing side of the optional wicking layer may be coupled to the environmental-facing side of the substrate sheet. In some embodiments, the wound-facing side of the sealing layer is coupled to the environmental-facing side of the wicking layer and/or the substrate sheet.

[00141] In any embodiment disclosed herein, the wound dressing is sterile and packaged in a microorganism-impermeable container.

[00142] In one aspect, the wound dressing of the present technology comprises a substrate sheet; a plurality of first microcapsules comprising a first biocompatible marker encapsulated by a first encapsulating material, wherein the first encapsulating material comprises a first collagen; and a plurality of second microcapsules comprising a second biocompatible marker encapsulated by a second encapsulating material, wherein the second encapsulating material comprises a gelatin and an ORC, and a third encapsulating material, wherein the third encapsulating material comprises a second collagen, wherein the third encapsulating material encapsulates the second encapsulating material; optionally, a wicking layer; and a sealing layer comprising a window for viewing the wicking layer and/or the substrate sheet. [00143] In one aspect, the present disclosure provides a wound dressing composition comprising a fdm of adhesive layer of a T.R.A.C.™ Pad; a plurality of first microcapsules, comprising a first biocompatible marker encapsulated by a first encapsulating material comprising a first collagen; a plurality of second microcapsules comprising a second biocompatible marker encapsulated by a second encapsulating material comprising a first encapsulating material comprising gelatin and an ORC and a third encapsulating material comprising a second collagen, wherein the third encapsulating material encapsulates the second encapsulating material.

[00144] In one aspect, the present disclosure provides a wound dressing composition comprising a wound-facing sheet, a substrate sheet; and a plurality of first microcapsules comprising a first biocompatible marker encapsulated by a first encapsulating material, wherein the first encapsulating material comprises a first collagen, wherein the plurality of first microcapsules are embedded in or coupled to the substrate sheet. Additionally or alternatively, in some embodiments, the wound dressing composition further comprises a wicking layer. Additionally or alternatively, in some embodiments, the wound dressing composition further comprises a sealing layer comprising a window for viewing the wicking layer and/or the substrate sheet. In this embodiment, the wound facing sheet contacts the wound bed. In some embodiments, the microcapsules do not directly contact the wound bed but contact wound exudate when the wound exudate reaches the substrate sheet by capillarity or because of negative pressure applied to drain the exudate. FIG. 3 shows an example of this embodiment.

[00145] FIG. 4 shows a diagrammatic representation of a wound dressing composition of the present technology, wherein the first and/or the second microcapsules of the present technology are concentrated in a patch area of the dressing. In some embodiments, the patch can be removed to see if a marker is released by the plurality of first and/or second microcapsules. This embodiment may include a window which can be used to view whether the microcapsules are degraded.

[00146] In one aspect, the present disclosure provides a wound dressing composition comprising a wound-facing sheet, a substrate sheet; and a plurality of second microcapsules comprising a second biocompatible marker encapsulated by a second encapsulating material, wherein the second encapsulating material comprises a gelatin and an ORC, and a third encapsulating material, wherein the third encapsulating material comprises a second collagen, wherein the third encapsulating material encapsulates the second encapsulating material, wherein the plurality of second microcapsules are embedded in or coupled to the substrate sheet. Additionally or alternatively, in some embodiments, the wound dressing composition further comprises a wicking layer. Additionally or alternatively, in some embodiments, the wound dressing composition further comprises a sealing layer comprising a window for viewing the wicking layer and/or the substrate sheet. In this embodiment, the wound facing sheet contacts the wound bed. The microcapsules do not directly contact the wound bed but contact wound exudate when the wound exudate reaches the substrate sheet by capillarity or because of negative pressure applied to drain the exudate. [00147] In one aspect, the present disclosure provides a wound dressing composition a wound facing sheet, a substrate sheet comprises a substrate sheet; a plurality of first microcapsules comprising a first biocompatible marker encapsulated by a first encapsulating material, wherein the first encapsulating material comprises a first collagen; and a plurality of second microcapsules comprising a second biocompatible marker encapsulated by a second encapsulating material, wherein the second encapsulating material comprises a gelatin and an ORC, and a third encapsulating material, wherein the third encapsulating material comprises a second collagen, wherein the third encapsulating material encapsulates the second encapsulating material; wherein the plurality of first microcapsules and the plurality of second microcapsules are embedded in or coupled to the substrate sheet. Additionally or alternatively, in some embodiments, the wound dressing composition further comprises a wicking layer. Additionally or alternatively, in some embodiments, the wound dressing composition further comprises a sealing layer comprising a window for viewing the wicking layer and/or the substrate sheet. In this embodiment, the wound-facing sheet contacts the wound bed. The microcapsules do not directly contact the wound bed but contact wound exudate when the wound exudate reaches the substrate sheet by capillarity or because of negative pressure applied to drain the exudate.

Diagnostic Methods of the Present Technology [00148] General. The wound dressing compositions of the present technology are useful in methods for detecting the activity levels of mediators of inflammation, including the proteases present in the wound bed (e.g., MMPs, HNE and bacterial proteases). The wound dressing compositions of the present technology are designed to release the biocompatible markers present in different type of microcapsules upon degradation of the encapsulating material of the microcapsules by the proteases present in the wound bed (e.g., MMPs, HNE and bacterial proteases). Accordingly, differing levels of release of different the biocompatible markers present in different type of microcapsules are helpful to diagnose conditions, including, but not limited to, inflammation, healing and infection.

[00149] In some cases, digestion of differing levels of different types of microcapsules may provide a colorimetric visualization that shows a range of activity of proteases present in the wound bed (e.g., MMPs, HNE and bacterial proteases) and may provide a threshold indication helpful to diagnose conditions such as inflammation, healing and/or infection. For example, when the first microcapsules encapsulate a yellow dye, the second microcapsules encapsulate a blue dye, and third microcapsules encapsulate a red dye, detection of yellow, red, green, orange, green, or purple color on the sealing layer may indicate activity of one or more type of proteases. Intensity of the color may indicate level of the activity. Such diagnosis may be helpful for the selection of a patient for additional therapy, such as of change of dressing, administration of one or more antibiotics, hospitalization, or a combination thereof.

[00150] Accordingly, in one aspect, the present disclosure provides a method of diagnosis of healing, inflammation, and/or infection of a wound, the method comprising (a) administering a device to the wound, wherein the device comprises the wound dressing of any embodiment disclosed herein, the apparatus of any embodiment disclosed herein, or the dressing system of any embodiment disclosed herein; (b) detecting change in color or fluorescence via a window for viewing the wicking layer and/or the substrate sheet; and (c) administering a therapeutic regimen when elevated levels of inflammatory mediators are detected compared to a predetermined threshold or a negative control sample. Additionally or alternatively, in some embodiments, the method further comprises applying the drape over the wound dressing and/or the retainer layer, wherein the drape is configured to seal the wound dressing of any of the embodiments disclosed herein, the apparatus of any of the embodiments disclosed herein or the system of any of the embodiments disclosed herein and/or the retainer layer and the wound site. Additionally or alternatively, in some embodiments, the method further comprises applying a retainer layer and/or a foam over the wound dressing. Additionally or alternatively, in some embodiments, the method further comprises applying the drape over the wound dressing and/or the retainer layer, wherein the drape is configured to seal the device and/or the retainer layer and the wound site and/or a vacuum for applying negative pressure to the wound, wherein the vacuum is configured to be fluidly connected to the drape through tubing. Additionally or alternatively, in some embodiments, the method further comprises optionally a vacuum for applying negative pressure to the wound, wherein the vacuum is configured to be fluidly connected to the drape through tubing. In some embodiments, the therapeutic regimen is selected from the group consisting of change of dressing, administration of one or more antibiotics, hospitalization, or a combination thereof.

Therapeutic Methods of the Present Technology

[00151] In one aspect, the present disclosure provides a method for treating a wound in a subject in need thereof, comprising administering to the wound the wound dressing of any embodiment disclosed herein, the apparatus of any embodiment disclosed herein, or the dressing system of any embodiment disclosed herein; optionally, applying a retainer layer and/or a foam over the wound dressing; optionally, applying the drape over the device and/or the retainer layer, wherein the drape is configured to seal the wound dressing and/or the retainer layer and the wound site; and optionally, applying negative pressure to the wound, wherein the vacuum is configured to be fluidly connected to the drape through tubing.

[00152] In one aspect, the present disclosure provides a method for treating a wound in a subject in need thereof, wherein the method may include administering to the wound a wound dressing composition of any embodiment disclosed herein. Additionally or alternatively in some embodiments, the wound dressing induces, enhances, or promotes healing in the subject. Additionally or alternatively, in some embodiments, the subject is human.

[00153] In one aspect, the present disclosure provides a method for protecting a wound from infection in a subject in need thereof, wherein the method may include administering to the wound a wound dressing composition of any embodiment disclosed herein. Additionally or alternatively in some embodiments, the wound dressing induces, enhances, or promotes healing in the subject. Additionally or alternatively, in some embodiments, the subject is human.

[00154] In any embodiment disclosed herein, the wound is an acute wound or a chronic wound. Additionally or alternatively, in some embodiments, 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, the wound is a chronic wound selected from the group consisting of infectious wounds, venous ulcers, arterial ulcers, decubitus ulcers and diabetic ulcers.

[00155] Any method known to those in the art for administering a wound dressing composition 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 dressings to a subject in need thereof, suitably a human. In any embodiment disclosed herein, the wound dressing may be applied directly to the wound. When used in vivo for therapy, the one or more wound dressings 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 used.

[00156] 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 dressings useful in the methods may be administered to a subject in need thereof by any number of well-known methods for administering wound dressings.

[00157] In any embodiment disclosed herein, the wound dressings 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 dressings are administered one, two, three, four, or five times per day. Additionally or alternatively, in some embodiments, the wound dressings are administered daily for one, two, three, four or five weeks. Additionally or alternatively, in some embodiments, the wound dressings are administered daily for less than 6 weeks. Additionally or alternatively, in some embodiments, the wound dressings are administered daily for 6 weeks or more. Additionally or alternatively, in some embodiments, the wound dressings are administered daily for 12 weeks or more. Additionally or alternatively, in some embodiments, the wound dressings 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 dressings are administered weekly, bi-weekly, tri-weekly, or monthly. Additionally or alternatively, in some embodiments, the wound dressings are administered for a period of one, two, three, four, or five weeks. Additionally or alternatively, in some embodiments, the wound dressings are administered for six weeks or more. Additionally or alternatively, in some embodiments, the wound dressings are administered for twelve weeks or more. Additionally or alternatively, in some embodiments, the wound dressings are administered for a period of less than one year. Additionally or alternatively, in some embodiments, the wound dressings are administered for a period of more than one year.

[00158] In some embodiments, the wound dressings 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, decubitus ulcers and diabetic ulcers.

[00159] In one aspect, the present disclosure provides a method for selecting a patient suspected of, or diagnosed as having an elevated level of inflammatory mediators in a wound for treatment with a therapeutic regimen comprising (a) administering a device to the wound, wherein the device comprises the wound dressing of any embodiment disclosed herein, the apparatus of any embodiment disclosed herein, or the system of any embodiment disclosed herein; (b) optionally, applying a retainer layer and/or a foam over the wound dressing; (c) optionally, applying the drape over the wound dressing and/or the retainer layer, wherein the drape is configured to seal the device and/or the retainer layer and the wound site; (d) optionally using a vacuum for applying negative pressure to the wound, wherein the vacuum is configured to be fluidly connected to the drape through tubing; (e) optionally, viewing the wicking layer and/or the substrate sheet through a window; (f) detecting change in color or fluorescence compared to a control sample; and (g) administering an appropriate therapeutic regimen when elevated levels of inflammatory mediators are detected compared to a predetermined threshold or a negative control sample. In some embodiments, the therapeutic regimen is selected from the group consisting of change of dressing, administration of one or more antibiotics, hospitalization, or a combination thereof.

Methods of Making the Wound Dressings of the Present Technology [00160] Also disclosed herein are methods for making the wound dressings of the present technology.

[00161] In one aspect, the present disclosure provides a method for making a wound dressing composition comprising: (a) providing (i) a substrate sheet; (ii) a plurality of first microcapsules comprising a first biocompatible marker encapsulated by a first encapsulating material, wherein the first encapsulating material comprises a first collagen; and (iii) a plurality of second microcapsules comprising a second biocompatible marker encapsulated by a second encapsulating material, wherein the second encapsulating material comprises a gelatin and an ORC, and a third encapsulating material, wherein the third encapsulating material comprises a second collagen, wherein the third encapsulating material encapsulates the second encapsulating material; (iv) optionally, a wicking layer; and (v) a sealing layer comprising a window for viewing the wicking layer and/or the substrate sheet; wherein the substrate sheet, the wicking layer, and the sealing layer independently comprise a wound-facing side and an environmental-facing side; (b) combining the substrate sheet, the wicking layer, and the sealing layer such that the wound-facing side of the sealing layer is coupled to the environmental- facing side of the wicking layer and/or the substrate sheet; and optionally, the wound-facing side of the wicking layer is coupled to the environmental-facing side of the substrate sheet; (c) coupling the plurality of first microcapsules with the wound-facing side of the substrate sheet; and (d) coupling the plurality of second microcapsules with the wound-facing side of the substrate sheet.

[00162] In one aspect, the present disclosure provides a method for making a wound dressing composition comprising: (a) providing (i) a substrate sheet; (ii) a plurality of first microcapsules comprising a first biocompatible marker encapsulated by a first encapsulating material, wherein the first encapsulating material comprises a first collagen; and (iii) a plurality of second microcapsules comprising a second biocompatible marker encapsulated by a second encapsulating material, wherein the second encapsulating material comprises a gelatin and an ORC, and a third encapsulating material, wherein the third encapsulating material comprises a second collagen, wherein the third encapsulating material encapsulates the second encapsulating material; (iv) optionally, a wicking layer; and (v) a sealing layer comprising a window for viewing the wicking layer and/or the substrate sheet; wherein the substrate sheet, the wicking layer, and the sealing layer independently comprise a wound-facing side and an environmental-facing side; (b) combining the substrate sheet, the wicking layer, and the sealing layer such that the wound-facing side of the sealing layer is coupled to the environmental- facing side of the wicking layer and/or the substrate sheet; and optionally, the wound-facing side of the wicking layer is coupled to the environmental-facing side of the substrate sheet; (c) coupling the first microcapsules with the wound-facing side of the substrate sheet; and (d) coupling the second microcapsules with the wound-facing side of the substrate sheet.

[00163] In one aspect, the present disclosure provides a method for making a wound dressing composition comprising: (a) providing (i) a substrate sheet; (ii) a plurality of first microcapsules comprising a first biocompatible marker encapsulated by a first encapsulating material, wherein the first encapsulating material comprises a first collagen; (iii) optionally, a wicking layer; and (iv) a sealing layer comprising a window for viewing the wicking layer and/or the substrate sheet; wherein the substrate sheet, the wicking layer, and the sealing layer independently comprise a wound-facing side and an environmental-facing side; (b) combining the substrate sheet, the wicking layer, and the sealing layer such that the wound-facing side of the sealing layer is coupled to the environmental- facing side of the wicking layer and/or the substrate sheet; and optionally, the wound-facing side of the wicking layer is coupled to the environmental-facing side of the substrate sheet; (c) coupling the first microcapsules with the wound-facing side of the substrate sheet; and (d) coupling the second microcapsules with the wound-facing side of the substrate sheet.

[00164] In one aspect, the present disclosure provides a method for making a wound dressing composition comprising: (a) providing (i) a substrate sheet; (ii) a plurality of second microcapsules comprising a second biocompatible marker encapsulated by a second encapsulating material, wherein the second encapsulating material comprises a gelatin and an ORC, and a third encapsulating material, wherein the third encapsulating material comprises a second collagen, wherein the third encapsulating material encapsulates the second encapsulating material; (iii) optionally, a wicking layer; and (iv) a sealing layer comprising a window for viewing the wicking layer and/or the substrate sheet; wherein the substrate sheet, the wicking layer, and the sealing layer independently comprise a wound-facing side and an environmental-facing side; (b) combining the substrate sheet, the wicking layer, and the sealing layer such that the wound-facing side of the sealing layer is coupled to the environmental- facing side of the wicking layer and/or the substrate sheet; and optionally, the wound-facing side of the wicking layer is coupled to the environmental-facing side of the substrate sheet; (c) coupling the first microcapsules with the wound-facing side of the substrate sheet; and (d) coupling the second microcapsules with the wound-facing side of the substrate sheet.

[00165] Additionally, or alternatively, in some embodiments, the first microcapsules and the second microcapsules have a diameter of from about 0.01 pm to about 1000 pm. In some embodiments, the first microcapsules and the second microcapsules have a diameter in a range independently selected from the group consisting of from about 0.01 pm to about 0.1 pm, from about 0.05 pm to about 0.25 pm, from about 0.1 pm to about 1 pm, from about 0.5 pm to about 2.5 pm, 1 pm to about 10 pm, from about 5 pm to about 25 pm, from about 10 pm to about 100 pm, from about 50 pm to about 250 pm, from about 100 pm to about 1000 pm, or any range including and/or in between any two of these values. In some embodiments, the first microcapsules and the second microcapsules have a diameter of about 5 pm to about 200 pm.

[00166] Additionally, or alternatively, in some embodiments, the first microcapsules and the second microcapsules have different sizes. In some embodiments, the first microcapsules are smaller than the second microcapsules.

[00167] Additionally, or alternatively, in some embodiments, the first microcapsules are digested faster by a matrix metalloproteinase (MMP) present in wound exudate compared to the second microcapsules.

[00168] Additionally, or alternatively, in some embodiments, the first collagen and the second collagen are independently recombinant or naturally occurring. Additionally, or alternatively, in some embodiments, the first collagen and the second collagen are mammalian collagens. In some embodiments, the first collagen and the second collagen independently comprise a bovine collagen, a human collagen, or a combination thereof. In some embodiments, the first collagen and/or second collagen comprises one or more of type I human recombinant collagen, type III human recombinant collagen, type I bovine collagen, type III bovine collagen, or a combination of any two or more thereof.

[00169] Additionally, or alternatively, in some embodiments, the first biocompatible marker and the second biocompatible marker independently comprise a dye or a fluorochrome. Additionally, or alternatively, in some embodiments, the first biocompatible marker and the second biocompatible marker are water soluble. Additionally, or alternatively, in some embodiments, the first biocompatible marker and/or the second biocompatible marker is substantially invisible when viewed using a light having wavelength of from about 380 nm to 740 nm. Additionally, or alternatively, in some embodiments, the first biocompatible marker and/or the second biocompatible marker is a pH indicator.

[00170] Additionally, or alternatively, in some embodiments, the first microcapsules and/or the second microcapsules are embedded in the substrate sheet. Additionally, or alternatively, in some embodiments, the substrate sheet comprises comprise a wound-facing side and an environmental- facing side. Additionally, or alternatively, in some embodiments, the first microcapsules and/or the second microcapsules are coupled to the wound-facing side of the substrate sheet. In some embodiments, the first microcapsules and/or the second microcapsules are coupled to the entire wound-facing side of the substrate sheet. Additionally, or alternatively, in some embodiments, the first microcapsules and/or the second microcapsules are coupled to only a part of the wound-facing side of the substrate sheet.

[00171] Additionally, or alternatively, in some embodiments, the wound dressing comprises about 0.01 wt.% to about 65 wt.%, more preferably about 0.01 wt % to about 10 wt%, of the first microcapsules. Additionally, or alternatively, in some embodiments, the wound dressing comprises about 0.01 wt.% to about 65 wt.%, more preferably about 0.01 wt % to about 10 wt%, of the second microcapsules.

[00172] Additionally, or alternatively, in some embodiments, the first microcapsules, the second microcapsules and/or the substrate sheet comprises a silver compound. In some embodiments, the first microcapsules, the second microcapsules and/or the substrate sheet comprises about 0.1 wt.% to about 3 wt.% of the silver compound. In some embodiments, the silver compound comprises one or more pharmaceutically acceptable silver salts. In some embodiments, the one or more pharmaceutically acceptable silver salts is selected from the group consisting of silver oxide, silver oxysalts, silver chromate, silver allantoinate, silver borate, silver glycerolate, silver nitrate, silver acetate, silver chloride, silver sulfate, silver lactate, silver bromide, silver iodide, silver carbonate, silver citrate, silver laurate, silver deoxycholate, silver salicylate, silver p-aminobenzoate, silver p- aminosalicylate, nanocrystalline silver, any pharmaceutically acceptable salt thereof, and any combination thereof.

[00173] Additionally, or alternatively, in some embodiments, the first microcapsules, the second microcapsules and/or the substrate sheet comprises further comprises one or more additional additives selected from the group consisting of an antimicrobial agent, an antioxidant, a signaling protein, and a combination any two or more thereof.

[00174] Additionally, or alternatively, in some embodiments, the first microcapsules, the second microcapsules and/or the substrate sheet comprises about 0.001 wt.% to about 5 wt.% of the antimicrobial agent. In some embodiments, the antimicrobial agent comprises one or more of tetracycline, penicillins, terramycins, erythromycin, bacitracin, neomycin, polymycin B, mupirocin, clindamycin, colloidal silver, silver sulfadiazine, chlorhexidine, povidone iodine, triclosan, sucralfate, quaternary ammonium salts, pharmaceutically acceptable silver salts, or any combination thereof. [00175] Additionally, or alternatively, in some embodiments, the first microcapsules, the second microcapsules and/or the substrate sheet comprises about 0.001 wt.% to about 5 wt.% of the antioxidant. In some embodiments, the antioxidant comprises one or more of anthocyanins, astaxanthin, bilirubin, canthaxanthin, capsaicin, citric acid, curcumin, coenzyme Q10, eugenol, flavanols, flavonolignans, flavanones, flavones, flavonols, iodide, isoflavone phytoestrogens, lutein, lycopene, manganese, melatonin, N-acetylcysteine, oxalic acid, phenolic acids, phytic acid, R-a-lipoic acid, stilbenoids, tocopherol, tocotrienol, vitamin A, vitamin C, vitamin E, xanthones, zeaxanthin, a- carotene, b-carotene, or any combination thereof. In some embodiments, the anthocyanins are selected from the group consisting of cyanidin, delphinidin, malvidin, pelargonidin, peonidin, petunidin, and any combination thereof. In some embodiments, the flavanols are selected from the group consisting of catechin, epicatechin, theaflavin, thearubigins, gallocatechin, epigallocatechin, or any gallate ester thereof, and any combination thereof. In some embodiments, the flavanones are selected from the group consisting of eriodictyol, hesperetin, naringenin, and any combination thereof. In some embodiment, the flavones are selected from the group consisting of apigenin, luteolin, tangeritin, and any combination thereof. In some embodiments, the flavonols are selected from the group consisting of isorhamnetin, kaempferol, myricetin, proanthocyanidins, quercetin, rutin, and any combination thereof. In some embodiments, the isoflavone phytoestrogens are selected from the group consisting of daidzein, genistein, glycitein, and any combination thereof. In some embodiments, 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. In some embodiments, the stilbenoids are selected from the group consisting of resveratrol, pterostilbene, and any combination thereof.

[00176] Additionally, or alternatively, in some embodiments, the first microcapsules, the second microcapsules and/or the substrate sheet comprises about 0.001 wt.% to about 5 wt.% of the signaling protein. In some embodiments, the signaling protein comprises one or more of platelet-derived growth factor (PDGF), transforming growth factor beta (TGF ), fibroblast growth factors (FGFs), epidermal growth factor (EGF), or any combination thereof. In some embodiments, the fibroblast growth factors (FGFs) 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. [00177] Additionally, or alternatively, in some embodiments, the first microcapsules and/or the second microcapsules are embedded in the substrate sheet. Additionally, or alternatively, in some embodiments, the substrate sheet comprises comprise a wound-facing side and an environmental- facing side. Additionally, or alternatively, in some embodiments, the first microcapsules and/or the second microcapsules are coupled to the wound-facing side of the substrate sheet. In some embodiments, the first microcapsules and/or the second microcapsules are coupled to the entire wound-facing side of the substrate sheet. Additionally, or alternatively, in some embodiments, the first microcapsules and/or the second microcapsules are coupled to only a part of the wound-facing side of the substrate sheet.

[00178] Additionally, or alternatively, in some embodiments, suitable reactive functional groups are added onto the first microcapsules and/or the second microcapsules for coupling to the substrate sheet. Additionally, or alternatively, in some embodiments, suitable reactive functional groups are added onto the substrate sheet for coupling to the first microcapsules and/or the second microcapsules. Non limiting examples of suitable reactive functional groups include groups such as acid anhydride groups, amino groups, N-substituted amino groups and their salts, epoxy groups (such as cyclohexyl epoxy groups), glycidyl groups, hydroxyl groups, isocyanate groups, urea groups, aldehyde groups, ester groups, ether groups, alkenyl groups, alkynyl groups, thiol groups, disulphide groups, silyl or silane groups, glyoxal-based groups, aziridine -based groups, groups based on active methylene compounds or other b-dicarbonyl compounds (such as 2,4-pentadione, malonic acid, acetylacetone, ethylacetone acetate, malonamide, acetoacetamide and its methyl analogues, ethyl acetoacetate and isopropyl acetoacetate), halo groups and hydrides. Polar groups (i.e. positively or negatively charged, zwitterionic or amphoteric groups) or hydrogen bonding groups may also be considered as reactive functional groups. The chemical bonds may be obtained through the introduction of functional groups in the first microcapsules, the second microcapsules and/or the substrate sheet that bind chemically to functional groups of a surface. The functional groups on the substrate may be selected or adapted to form a chemical bond, with the functional groups on the first microcapsules and/or the second microcapsules. The chemical bonds can be ionic, hydrogen bonding or, better still, covalent, where a simple chemical reaction takes place by addition or substitution. Reaction may be promoted solely by the pH of the solution in which the microcapsules are applied to the surface, normally an alkaline solution, or initiators may be included in case of an addition radical reaction.

[00179] Additionally, or alternatively, in some embodiments, suitable reactive functional groups may be added to the substrate sheet in form of trace amount of fibers having functional groups. Additionally, or alternatively, in some embodiments, suitable reactive functional groups may be introduced onto the outer surface of the first microcapsules and/or the second microcapsules, for instance by adding trace amounts of one or more chemical to the encapsulating material of any embodiment disclosed herein. For example, the encapsulating material of any embodiment disclosed herein may include a melamine formaldehyde resin but with the polymerization process controlled in terms of temperature, catalyst and pH such that not all amino groups of the melamine are reacted, leaving free primary and secondary groups on the outer face of the microcapsules, which may be verified by acid-base titration. The reactive functional groups may be introduced into the outer surface of the first microcapsules and/or the second microcapsules via the urea-formaldehyde, melamine-formaldehyde, polyamide or chitosan included in the encapsulating material of any embodiment disclosed herein by reaction with the amino or hydroxyl groups present on the outer faces of such microcapsules. Glycidyl methacrylate or any other suitable monomer that may contain epoxy (glycidyl) groups, or acrylic acid containing carboxylic groups may be used. Other suitable reactive functional groups are listed hereinbefore.

[00180] Additionally, or alternatively, in some embodiments, the first microcapsules and/or the second microcapsules may include reactive functional groups on the outer surface of the first microcapsules and/or the second microcapsules that convey affinity towards the substrate sheet or can react with the hydroxyl groups of the cellulose, if present in the substrate sheet. Non-limiting examples of such reactive functional groups are epoxy group, alkyl with a halogen substitution, such as ethyl chlorine, vinyl groups or heterocyclic groups. Other suitable reactive functional groups are listed hereinbefore. Where the outer surface the first microcapsules and/or the second microcapsules contains an aminoplast resin such as urea-formaldehyde or melamine- formaldehyde, or a polyamide or chitosan, the introduction of functional groups, such as epoxy groups or ethyl chlorine, for example, may be achieved through a reaction between unreacted free amine groups and a bifunctional bridging agent (i.e. bonding agent) that contains epoxy groups, alkyl groups substituted with a halogens vinyl groups or heterocyclic, leaving the other group of the bifunctional agent free for reacting with the surface. Additional details of microcapsules with functional reactive groups for binding to surfaces, specifically to fibers, may be found in the publication WO 2006/117702, which is incorporated herein in its entirety.

[00181] Additionally, or alternatively, in some embodiments, having the functional groups and/or by contacting the first microcapsules, the second microcapsules and/or the substrate sheet with a chemical having a suitable reactive functional group. Additionally, or alternatively, in some embodiments, the process of introducing suitable reactive functional groups in the substrate sheet may be similar to the dyeing process with reactive dyes. Additionally, or alternatively, in some embodiments, the process of introducing suitable reactive functional groups the outer surface of the first microcapsules and/or the second microcapsules may be similar to the dyeing process with reactive dyes. The suitable reactive functional groups are listed hereinbefore.

[00182] Additionally, or alternatively, in some embodiments, the functional reactive groups located on the outer surface of the first microcapsules and/or the second microcapsules may be adapted to react with the functional reactive groups of the substrate sheet, whereby a covalent bond is formed between the functional reactive groups of the first microcapsules and/or the second microcapsules, and the substrate sheet. Kits Comprising the Wound Dressings of the Present Technology [00183] 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.

EXAMPLES

[00184] 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 : Wound Dressings of the Present Technology Detect Protease Activity [00185] A colorimetric assay will be utilized to determine the protease activities after incubation with the test wound dressing samples. Briefly, a wound dressing compositions of the present technology will be applied to a wound model irrigated with 0.5 ml/ hr of (a) phosphate buffered saline, (b) a solution of simulated wound fluid (SWF) containing human neutrophil elastase (FINE, 273 mU/mF), (3) or SWF containing matrix metalloproteinase-9 (MMP-9, 1 pg/mF), or (4) SWF containing neutrophil elastase (FINE, 273 mU/mF) and matrix metalloproteinase-9 (MMP-9, 1 pg/mF). The wound dressing composition will be incubated for 2 hours at 37° C. After 2 hr, the color change of the wicking layer and/or the substrate sheet will be determined.

[00186] Another set of wound dressing compositions of the present technology will be will be applied to a wound model irrigated with 0.5 ml/ hr of (a) phosphate buffered saline, (b) a solution of simulated wound fluid (SWF) containing human neutrophil elastase (FINE, 273 mU/mF), (3) or SWF containing matrix metalloproteinase-9 (MMP-9, 1 pg/mF), or (4) SWF containing neutrophil elastase (FINE, 273 mU/mF) and matrix metalloproteinase-9 (MMP-9, 1 pg/mF). These wound dressings will be incubated for 24 hours at 37° C, and the color change of the wicking layer and/or the substrate sheet will be determined.

[00187] It is anticipated that the wound dressings of the present technology will exhibit the color released by the first microcapsules when treated with a solution of simulated wound fluid (SWF) containing human neutrophil elastase (FINE, 273 mU/mF). It is also anticipated that the wound dressings of the present technology will exhibit the color released by the second microcapsules when treated with a solution of SWF containing matrix metalloproteinase-9 (MMP-9, 1 pg/mF). It is further anticipated that the wound dressings of the present technology will exhibit the colors released by the first and second microcapsules when treated with SWF containing neutrophil elastase (FINE, 273 mU/mL) and matrix metalloproteinase-9 (MMP-9, 1 pg/mL). The intensity of the color is anticipated to increase after 24 hr.

[00188] Accordingly, the wound dressing compositions of the present technology are useful in methods for treating a wound in a subject in need thereof, wherein the method comprises administering to the wound a dressing of any embodiment disclosed herein.

EQUIVALENTS

[00189] 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.

[00190] 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.

[00191] As will be understood by one skilled in the art, for any and all purposes, particularly in terms of providing a written description, all ranges disclosed herein also encompass any and all possible subranges and combinations of subranges thereof. Any listed range can be easily recognized as sufficiently describing and enabling the same range being broken down into at least equal halves, thirds, quarters, fifths, tenths, etc. As a non-limiting example, each range discussed herein can be readily broken down into a lower third, middle third, and upper third, etc. As will also be understood by one skilled in the art all language such as “up to,” “at least,” “greater than,” “less than,” and the like, include the number recited and refer to ranges which can be subsequently broken down into subranges as discussed above. Finally, as will be understood by one skilled in the art, a range includes each individual member. Thus, for example, a group having 1-3 cells refers to groups having 1, 2, or 3 cells. Similarly, a group having 1-5 cells refers to groups having 1, 2, 3, 4, or 5 cells, and so forth.

[00192] All patents, patent applications, provisional applications, and publications referred to or cited herein are incorporated by reference in their entirety, including all figures and tables, to the extent they are not inconsistent with the explicit teachings of this specification.

Claims

1. A wound dressing composition comprising: a substrate sheet; a plurality of first microcapsules comprising a first biocompatible marker encapsulated by a first encapsulating material, wherein the first encapsulating material comprises a first collagen; and a plurality of second microcapsules comprising a second biocompatible marker encapsulated by a second encapsulating material, and a third encapsulating material, wherein the second encapsulating material comprises a gelatin and an oxidized regenerated cellulose (ORC), wherein the third encapsulating material comprises a second collagen, and wherein the third encapsulating material encapsulates the second encapsulating material.

2. The wound dressing composition of claim 1 further comprising a wicking layer.

3. The wound dressing composition of claim 1 or claim 2 further comprising a sealing layer comprising a window for viewing the wicking layer and/or the substrate sheet.

4. The wound dressing composition of any one of claims 1-3, wherein the plurality of first microcapsules and the plurality of second microcapsules have a diameter of from about 0.01 pm to about 1000 pm.

5. The wound dressing composition of any one of claims 1-4, wherein the plurality of first microcapsules and the plurality of second microcapsules have a diameter of about 5 pm to 200 pm.

6. The wound dressing composition of any one of claims 1-5, wherein the plurality of first microcapsules are digested faster by a matrix metalloproteinase (MMP) present in wound exudate compared to the plurality of second microcapsules.

7. The wound dressing composition of any one of claims 1-6, wherein the first collagen and the second collagen are independently recombinant or naturally occurring collagen.

8. The wound dressing composition of any one of claims 1-7, wherein the first collagen and/or second collagen comprises one or more of type I human recombinant collagen, type III human recombinant collagen, type I bovine collagen, type III bovine collagen, or a combination of any two or more thereof.

9. The wound dressing composition of any one of claims 1-8, wherein the first biocompatible marker and the second biocompatible marker independently comprise a dye or a fluorochrome.

10. The wound dressing composition of any one of claims 1-9, wherein the substrate sheet comprises comprise a wound-facing side and an environmental -facing side.

11. The wound dressing composition of claim 10, wherein the plurality of first microcapsules and the plurality of second microcapsules are coupled to the wound-facing side of the substrate sheet.

12. The wound dressing composition of any one of claims 1-11, wherein the wound dressing comprises about 0.01 wt.% to about 65 wt.%, more preferably about 0.01 wt % to about 10 wt%, of the plurality of first microcapsules and about 0.01 wt.% to about 65 wt.% .%, more preferably about 0.01 wt % to about 10 wt%, of the plurality of second microcapsules.

13. The wound dressing composition of any one of claims 1-12, wherein the plurality of first microcapsules, the plurality of second microcapsules and/or the substrate sheet comprise a silver compound.

14. The wound dressing composition of claim 13, wherein the silver compound comprises one or more pharmaceutically acceptable silver salt selected from the group consisting of silver oxide, silver oxysalts, silver chromate, silver allantoinate, silver borate, silver glycerolate, silver nitrate, silver acetate, silver chloride, silver sulfate, silver lactate, silver bromide, silver iodide, silver carbonate, silver citrate, silver laurate, silver deoxycholate, silver salicylate, silver p- aminobenzoate, silver /^-aminosalicylate. nanocrystalline silver, any pharmaceutically acceptable salt thereof, and any combination thereof.

15. The wound dressing composition of any one of claims 1-14, wherein the first microcapsule, the second microcapsule and/or the substrate sheet further comprises one or more additional additives selected from the group consisting of an antimicrobial agent, an antioxidant, a signaling protein, or a combination any two or more thereof.

16. A wound dressing composition comprising: a substrate sheet; and a plurality of microcapsules comprising a biocompatible marker encapsulated by a an encapsulating material, wherein the encapsulating material comprises a collagen.

17. The wound dressing composition of claim 16 further comprising a wicking layer.

18. The wound dressing composition of claim 16 or claim 17 further comprising a sealing layer comprising a window for viewing the wicking layer and/or the substrate sheet.

19. A wound dressing composition comprising: a substrate sheet; and a plurality of microcapsules comprising a biocompatible marker encapsulated by a first encapsulating material, and a second encapsulating material, wherein the first encapsulating material comprises a gelatin and an oxidized regenerated cellulose (ORC), wherein the second encapsulating material comprises a collagen, wherein the second encapsulating material encapsulates the first encapsulating material.

20. The wound dressing composition of claim 19 further comprising a wicking layer.

21. The wound dressing composition of claim 19 or claim 20 further comprising a sealing layer comprising a window for viewing the wicking layer and/or the substrate sheet.

22. An apparatus comprising: a substrate sheet; a plurality of first microcapsules comprising a first biocompatible marker encapsulated by a first encapsulating material, wherein the first encapsulating material comprises a first collagen; and a plurality of second microcapsules comprising a second biocompatible marker encapsulated by a second encapsulating material, and a third encapsulating material wherein the second encapsulating material comprises a gelatin and an ORC, wherein the third encapsulating material comprises a second collagen, wherein the third encapsulating material encapsulates the second encapsulating material.

23. The apparatus of claim 22 further comprising a wicking layer.

24. The apparatus of claim 22 or claim 23 further comprising a sealing layer comprising a window for viewing the wicking layer and/or the substrate sheet.

25. A system comprising: a negative pressure source; and the wound dressing composition of any one of claims 1-21, or the apparatus of any one of claims 22-24.

26. A wound dressing composition comprising a wound-facing sheet, a substrate sheet; and a plurality of microcapsules comprising a biocompatible marker encapsulated by an encapsulating material, wherein the encapsulating material comprises a collagen, wherein the plurality of microcapsules are embedded in or coupled to the substrate sheet.

27. The wound dressing composition of claim 26 further comprising a wicking layer.

28. The wound dressing composition of claim 26 or claim 27 further comprising a sealing layer comprising a window for viewing the wicking layer and/or the substrate sheet.

29. A method for treating a wound in a subject in need thereof, comprising: administering to the wound the wound dressing composition of any one of claims 1-21 or 26- 28, the apparatus of any one of claims 22-24, or the system of claim 25.

30. The method of claim 29 further comprising:

(a) applying a retainer layer and/or a foam over the wound dressing composition or the apparatus;

(b) applying a drape over the wound dressing composition or the apparatus, and/or the retainer layer, wherein the drape is configured to seal the wound dressing composition, or the apparatus, and/or the retainer layer and the wound site; and

(c) applying negative pressure to the wound, wherein the vacuum is configured to be fluidly connected to the drape through tubing.

31. A method for selecting a patient suspected of, or diagnosed as having an elevated levels of inflammatory mediators in a wound for treatment with a therapeutic regimen comprising:

(a) administering a device to the wound, wherein the device comprises: the wound dressing composition of any one of claims 1-21 or 26-28, the apparatus of any one of claims 22-24, or the system claim 25;

(c) detecting change in color or fluorescence via the window for viewing the wicking layer and/or the substrate sheet; and (d) administering a therapeutic regimen when elevated levels of inflammatory mediators are detected compared to a predetermined threshold or a negative control sample.

32. The method of claim 31, wherein the therapeutic regimen is selected from the group consisting of change of dressing, administration of one or more antibiotics, hospitalization, or a combination thereof.

33. A method for making a wound dressing composition comprising:

(a) providing

(i) a substrate sheet;

(ii) a plurality of first microcapsules comprising a first biocompatible marker encapsulated by a first encapsulating material, wherein the first encapsulating material comprises a first collagen; and

(iii) a plurality of second microcapsules comprising a second biocompatible marker encapsulated by a second encapsulating material, and a third encapsulating material wherein the second encapsulating material comprises a gelatin and an ORC, wherein the third encapsulating material comprises a second collagen, wherein the third encapsulating material encapsulates the second encapsulating material;

(iv) a wicking layer; and

(v) a sealing layer comprising a window for viewing the wicking layer and/or the substrate sheet wherein each of the substrate sheet, the wicking layer, and the sealing layer independently comprise a wound-facing side and an environmental-facing side;

(b) combining the substrate sheet, the wicking layer, and the sealing layer such that the wound-facing side of the sealing layer is coupled to the environmental-facing side of the wicking layer and/or the substrate sheet; and optionally, the wound-facing side of the wicking layer is coupled to the environmental-facing side of the substrate sheet;

(c) coupling the plurality of first microcapsules and the plurality of second microcapsules with the wound-facing side of the substrate sheet.

34. A kit comprising the wound dressing composition of any one of claims 1-21, or 26-28, the apparatus of any one of claims 22-24 or 26-28, or the system of claim 25, and instructions for use.