TOPICAL COMPOSITION FOR USE IN THE TREATMENT OF BURNS

Abstract

The present disclosure relates to topical compositions for the treatment or prophylaxis of burns. The topical compositions comprise water, solvent, thickener, preservative and conditioning agent, and are exposed to gamma radiation. One embodiment of the topical compositions includes water, propanediol, sodium acryloyldimethyltaurate/VP crosspolymer, a combination of phenoxyethanol and caprylyl glycol and chlorphenesin, polyaminopropyl biguanide (PHMB) and a mineral complex. The present disclosure also relates to the use of the topical compositions with a dressing and the use of the topical compositions and dressings in treatment or prophylaxis of burns.

Claims

1. A topical composition comprising: water, in a range of approximately 85-95% w/w of the composition; propanediol, in a range of approximately 5-10% w/w of the composition; sodium acryloyldimethyltaurate/VP crosspolymer, in a range of approximately 0.5-1.0% w/w of the composition; one or more preservatives, in a range of approximately 0.5-2.0% w/w of the composition; and a mineral complex, in a range of approximately 0.1-1.0% w/w of the composition.

2. The topical composition of claim 1, wherein the one or more preservatives is selected from the group consisting of polyaminopropyl biguanide (PHMB) and a combination of phenoxyethanol and caprylyl glycol and chlorphenesin.

3. The topical composition of claim 1, wherein the composition has a viscosity approximately in the range 200-6000 cP at 25° C. following exposure to gamma radiation.

4. The topical composition of claim 1, wherein the water is selected from the group consisting of tap water, purified water, sterile water and halogenated water.

5. The topical composition of claim 1 wherein the mineral complex is sea water extract.

6. The topical composition of claim 1 wherein the mineral complex comprises magnesium, potassium, sodium, boron and calcium.

7. The topical composition of claim 6 wherein the mineral complex further comprises one or more from the group consisting of: copper, nickel, silicon, zinc, aluminium, arsenic, barium, cadmium, cobalt, chromium, iron, mercury, manganese, lead, antimony, selenium, tin, strontium, titanium and vanadium.

8. The topical composition of claim 1, wherein the composition has a specific gravity of approximately 1.000±0.05 at 25° C.

9. The topical composition of claim 1, wherein the composition has a pH approximately in the range 4.0-6.5 at 25° C.

10. A burn dressing comprising the topical composition of claim 1 and a dressing material.

11. A method of sterilising a topical composition of claim 1 comprising applying gamma radiation of approximately 25.0 to 44.5 kGy to the composition.

12. The composition of claim 1 which has been sterilised using the method of claim 1.

13. A method of prophylaxis or treatment of a burn comprising the step of applying the topical composition of claim 1 to skin in need thereof.

14. The burn dressing according to claim 10 which has been sterilised using the method of claim 11.

15. A method of prophylaxis or treatment of a burn comprising the step of applying a burn dressing according to claim 10 to skin in need thereof.

16. A topical composition consisting of: water, in a range of approximately 85-95% w/w of the composition; propanediol, in a range of approximately 5-10% w/w of the composition; sodium acryloyldimethyltaurate/VP crosspolymer, in a range of approximately 0.5-1.0% w/w of the composition; a combination of phenoxyethanol and caprylyl glycol and chlorphenesin, in a range of approximately 0.5-1.5% w/w of the composition; polyaminopropyl biguanide (PHMB), in a range of approximately 0.05-0.15% w/w of the composition; and a mineral complex, in a range of approximately 0.1-1.0% w/w of the composition.

17. The topical composition of claim 16, wherein the composition has a viscosity approximately in the range 200-6000 cP at 25° C. following exposure to gamma radiation of approximately 25.0 to 44.5 kGy.

18. A burn dressing comprising the topical composition of claim 16 and a dressing material.

19. A method of prophylaxis or treatment of a burn comprising the step of applying the topical composition of claim 16 to skin in need thereof.

20. The topical composition according to claim 16, wherein the mineral complex comprises magnesium, potassium, sodium, boron, calcium and optionally one or more from the group consisting: copper, nickel, silicon, zinc, aluminium, arsenic, barium, cadmium, cobalt, chromium, iron, mercury, manganese, lead, antimony, selenium, tin, strontium, titanium and vanadium.

Description

BRIEF DESCRIPTION OF THE FIGURES

[0034] To further clarify the above and other advantages and features of the present invention, a more particular description of the invention will be rendered by reference to specific embodiments thereof which are illustrated in the appended drawings. It is appreciated that these drawings depict only typical embodiments of the invention and are therefore not to be considered limiting of its scope.

[0035] FIG. 1 shows the results of a wound healing assay—Human Primary Dermal Fibroblast data plot Cell index v time.

[0036] FIG. 2 shows Human Primary Keratinocyte cells cell index v time.

[0037] FIG. 3 shows RT2 qPCR of fibroblast monoculture comparing cells exposed to the composition versus a control (untreated cells).

[0038] FIG. 4 shows a representation of the LabSkin system including a cross section through the striated skin. Well insert contains cultured cells in 3D fibrin scaffold.

[0039] FIG. 5a shows the brass weights that we employed in inflicting thermal burn injury and FIG. 5b shows the location of subsequent skin biopsies following burn injury.

[0040] FIGS. 6a and 6b show six samples (numbered 1-6) of the damaged (burned) skin 24 hours after burn burns were inflicted.

[0041] FIG. 7a shows tissue dielectric constant (TDC) as an index of localised skin water content in control model (FIG. 7a) and when treated with mineral complex (FIG. 7b).

[0042] FIG. 8a shows wound healing PCR arrays revealing up- and down-regulated genes in 3D skin models in response to thermal burn injury (no treatment) vs healthy skin. Total RNA from 3D skin models were characterised, and the relative expression levels for each gene in the two samples (burn vs healthy skin) are plotted against each other in the Scatter Plot.

[0043] FIG. 8b shows wound healing PCR arrays revealing up- and down-regulated genes in 3D skin models in response to treatment with NB105-146 (gel formulation without mineral complex) for thermal burn injury. Total RNA from 3D skin models were characterised, and the relative expression levels for each gene in the two samples (treated vs burn (untreated) skin) are plotted against each other in the Scatter Plot.

[0044] FIG. 8c shows wound healing PCR arrays revealing up- and down-regulated genes in 3D skin models in response to treatment with NB105-142 (gel formulation with mineral complex) for thermal burn injuries. Total RNA from 3D skin models were characterised, and the relative expression levels for each gene in the two samples (treated vs Burn (untreated) skin) are plotted against each other in the Scatter Plot.

DESCRIPTION

[0045] Burn as employed herein means an injury to skin, or other tissues, caused by heat, cold, electricity, chemicals, friction, or radiation. Compositions of the present disclosure are particularly beneficial in the treatment and prophylaxis of thermal and radiation burns although they can be employed in the treatment of any burn, including chemical burns.

[0046] In one embodiment the composition is suitable for the treatment or prophylaxis of burns, such as thermal or radiation burns, particularly thermal burns.

[0047] In one embodiment there is provided a composition for use in the treatment or prophylaxis of burns, such as thermal or radiation burns, particularly thermal burns.

[0048] As employed herein thermal burns refers to burns that are not chemical or radiation burns.

[0049] In one embodiment there is provided a composition for use in the prophylaxis of radiation burns.

[0050] Prophylaxis as employed herein refers to the prevention of condition aimed at stopping the condition developing or progressing, such as a burn or burns.

[0051] Treatment as employed herein refers to the reversal of a condition, amelioration or relief of symptoms associated with a condition or prevention of further development/worsening of a condition, such as a burn or burns.

[0052] Composition

[0053] In one embodiment there is provided a topical composition comprising water, solvent, thickener, preservative and mineral complex conditioning agent wherein the composition has a viscosity approximately in the range 200-6000 cP at 25° C. following exposure to gamma radiation.

[0054] Topical composition as employed herein means preparation that is applied to the surface of the body, such as the skin, including but not limited to a cream, foam, ointment, paste, lotion or gel, including a hydrogel.

[0055] In one embodiment the topical composition is a fluid or a gel.

[0056] Water as employed herein typically refers to purified water that has been cleaned and/or filtered to be suitable for topical application. Water may refer to tap water, purified water, sterile water, halogenated water (especially chlorinated water), and mixtures thereof. As employed herein, water has a heat-absorbing function, aimed at cooling the sensation of heat in the skin following a burn. The water also acts as a solvent. Water as employed herein has the CAS number 7732-18-5 as defined by the chemical abstract service.

[0057] In one embodiment the water is purified water. In one embodiment the water is present at approximately 85-95% w/w of the total composition, such as approximately 85.5, 86, 86.5, 87, 87.5, 88, 88.5, 89.5, 90, 90.5, 91, 91.5, 92, 92.5, 93, 93.5, 94 or 94.5% w/w of the total composition, for example approximately 89.45% w/w of the total composition. In one embodiment, the balance of the composition, following addition of other components, is water.

[0058] Solvent as employed herein means a substance (a liquid) that dissolves a solute (a chemically distinct liquid, solid or gas), resulting in a solution.

[0059] In one embodiment the solvent is present at approximately 5-10% w/w of the total composition, such as approximately 6, 7, 8 or 9% w/w of the total composition, for example approximately 8% w/w of the total composition.

[0060] In one embodiment the solvent is propanediol. In one embodiment the propanediol comprises approximately 5-10% w/w of the total composition, such as approximately 6, 7, 8 or 9% w/w of the total composition, for example approximately 8% w/w of the total composition.

[0061] Propanediol as employed herein means 1,3-propanediol, a chemical according to formula (I)

##STR00001##

[0062] Propanediol as employed herein has the CAS number 504-63-2.

[0063] Thickener or thickening agent as employed herein is an ingredient or ingredients that increase the viscosity of a composition without substantially altering its other properties. Examples of thickening agents include polysaccharides such as gums, starches, in particular corn starch, carbomers, gelling agents and acrylates such as sodium acryloyldimethyltaurate/VP crosspolymer (Aristoflex AVS®).

[0064] In one embodiment the thickener comprises approximately 0.5-1.0% w/w of the total composition, such as approximately 0.55, 0.6, 0.65, 0.7, 0.75, 0.8, 0.85, 0.9 or 0.95% w/w of the total composition, for example approximately 0.8% w/w of the total composition.

[0065] In one embodiment the thickener is sodium acryloyldimethyltaurate/VP crosspolymer. Sodium acryloyldimethyltaurate/VP crosspolymer as employed herein has the CAS number 1176663-96-9. In one embodiment the sodium acryloyldimethyltaurate/VP crosspolymer comprises approximately 0.5-1.0% w/w of the total composition, such as approximately 0.55, 0.6, 0.65, 0.7, 0.75, 0.8, 0.85, 0.9 or 0.95% w/w of the total composition, for example approximately 0.8% w/w of the total composition.

[0066] Preservative as employed herein refers to a substance that prevents decomposition or contamination either by microorganisms or by chemical change. Typical preservatives suitable for topical compositions include, but are not limited to, phenoxyethanol, ethylhexylglycerine, caprylyl glycol, chlorphenesin, quaternary ammonium compounds, such as benzalkonium chloride, benzethonium chloride, cetrimide, dequalinium chloride, and cetylpyridinium chloride; mercurial agents, such as phenylmercuric nitrate, phenylmercuric acetate, and thimerosal; alcoholic agents, for example, chlorobutanol, phenylethyl alcohol, and benzyl alcohol; antibacterial esters, other examples include, esters of parahydroxybenzoic acid; and other anti-microbial agents such as chlorhexidine, chlorocresol, benzoic acid and polymyxin.

[0067] In one embodiment the preservative comprises approximately 0.5-2.0% w/w of the total composition, such as approximately 0.55, 0.6, 0.65, 0.7, 0.75, 0.8, 0.85, 0.9, 0.95, 1.0, 1.05, 1.1, 1.15, 1.2, 1.25, 1.3, 1.35, 1.4, 1.45, 1.5, 1.55, 1.6, 1.65, 1.7, 1.75, 1.8, 1.85, 1.9 or 1.95% w/w of the total composition, for example approximately 1.5% w/w of the total composition.

[0068] In one embodiment the composition comprises one or more preservatives from the group consisting of: phenoxyethanol and caprylyl glycol and chlorphenesin (commercially known as Mikrokill®COS) and (PHMB) polyaminopropyl biguanide.

[0069] In one embodiment the preservative is phenoxyethanol and caprylyl glycol and chlorphenesin (Mikrokill®) and (PHMB) polyaminopropyl biguanide.

[0070] In one embodiment the phenoxyethanol and caprylyl glycol and chlorphenesin (Mikrokill®COS) comprises approximately 0.5-1.5% w/w of the total composition, such as approximately 0.55, 0.6, 0.65, 0.7, 0.75, 0.8, 0.85, 0.9, 0.95, 1.0, 1.05, 1.1, 1.15, 1.2, 1.25, 1.3, 1.35, 1.4 or 1.45% w/w of the total composition, for example approximately 1.0% w/w of the total composition.

[0071] As employed herein phenoxyethanol & caprylyl glycol & chlorphenesin is the INCI name for Mikrokill®COS and has the CAS number 122-99-6/1117-86-8/104-29-0.

[0072] The composition may comprise approximately 0.25-0.75% (PHMB) polyaminopropyl biguanide, in particular approximately 0.5% (PHMB) polyaminopropyl biguanide.

[0073] The composition may comprise approximately 0.05-0.15% (PHMB) polyaminopropyl biguanide, in particular approximately 0.1% (PHMB) polyaminopropyl biguanide.

[0074] In one embodiment the (PHMB) polyaminopropyl biguanide comprises approximately 0.25-0.75% w/w of the total composition, such as 0.3, 0.35, 0.4, 0.45, 0.5, 0.55, 0.6, 0.65 or 0.7% w/w of the total composition, for example approximately 0.5% w/w of the total composition. (PHMB) polyaminopropyl biguanide as employed herein has the CAS number 133029-32-0/27083-27-8. polyaminopropyl biguanide is the INCI name. PHMB (polyhexamethylene biguanide) is the chemical name. In one embodiment the (PHMB) polyaminopropyl biguanide is provided as a 20% solution, thus 0.5% of the solution contains 0.1% (PHMB) polyaminopropyl biguanide on a pure basis.

[0075] In one embodiment the (PHMB) polyaminopropyl biguanide comprises approximately 0.05-0.15% w/w of the total composition, such as 0.06, 0.07, 0.08, 0.09, 0.1, 0.11, 0.12, 0.13 or 0.14% w/w of the total composition, for example approximately 0.1% w/w of the total composition. (PHMB) polyaminopropyl biguanide as employed herein has the CAS number 133029-32-0/27083-27-8. polyaminopropyl biguanide is the INCI name. PHMB (polyhexamethylene biguanide) is the chemical name. Typically, the (PHMB) polyaminopropyl biguanide is provided as a 20% solution, thus 0.1% of the solution contains 0.02% (PHMB) polyaminopropyl biguanide on a pure basis.

[0076] In one embodiment there is provided a topical composition comprising approximately 1.0% w/w phenoxyethanol and caprylyl glycol and chlorphenesin plus an additional approximately 0.5% w/w (PHMB) polyaminopropyl biguanide (20% solution).

[0077] In one embodiment there is provided a topical composition comprising approximately 1.0% w/w phenoxyethanol and caprylyl glycol and chlorphenesin plus an additional approximately 0.1% w/w (PHMB) polyaminopropyl biguanide (20% solution).

[0078] Mineral complex conditioning agent as employed herein means an agent designed to improve the condition of the skin.

[0079] In one embodiment the mineral complex conditioning agent comprises approximately 0.1-1.0% w/w of the total composition, such as approximately 0.15, 0.2, 0.25, 0.3, 0.35, 0.4, 0.45, 0.5, 0.55, 0.6, 0.65, 0.7, 0.75, 0.8, 0.85, 0.9 or 0.95% w/w of the total composition, for example approximately 0.25% w/w of the total composition.

[0080] In some embodiments the conditioning agent is a mineral complex. In one embodiment the mineral complex comprises approximately 0.1-1.0% w/w of the total composition, such as approximately 0.15, 0.2, 0.25, 0.3, 0.35, 0.4, 0.45, 0.5, 0.55, 0.6, 0.65, 0.7, 0.75, 0.8, 0.85, 0.9 or 0.95% w/w of the total composition, for example approximately 0.25% w/w of the total composition.

[0081] Mineral complex as employed herein refers to a complex of several minerals, typically including, but not limited to magnesium, potassium, sodium, boron, calcium. The conditioning agent/mineral complex is described in further detail below.

[0082] Viscosity as employed herein is a measure of a fluid's resistance to flow. It corresponds to a notional “thickness” of a liquid and is measured in cP (centipoise). Centipoise is a measure of viscosity on the CGS (centimetre gram second) scale. Water has a viscosity of 1 cP at 20° C. Viscosity can be measured using a Brookfield viscometer, such as a Brookfield DV II Pro. Generally, viscosity is measured at room temperature, such as 20 to 25° C., preferably 25° C.

[0083] In one embodiment there is provided a topical composition with a viscosity (at approximately 25° C.) in the range approximately 100 to 6000 cP, such as approximately 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 950, 1000, 1100, 1200, 1300, 1400, 1500, 1600, 1700, 1800, 1900, 2000, 2100, 2200, 2300, 2400, 2500, 2600, 2700, 2800, 2900, 3000, 3100, 3200, 3300, 3400, 3500, 3600, 3700, 3800, 3900, 4000, 4100, 4200, 4300, 4400, 4500, 4600, 4700, 4800, 4900, 5000, 5100, 5200, 5300, 5400, 5500, 5600, 5700, 5800 or 5900 cP, for example approximately 200-6000 cP.

[0084] In one embodiment the composition has a viscosity in the range 200 to 6000 cP measure using spindle #63 spindle @ 12 RPM.

[0085] As employed herein, in relation to the constituents of the composition, all % are % w/w of the total composition.

[0086] Exposure to gamma radiation as employed herein means exposure to electromagnetic radiation typically having energy above 100 keV, frequencies above 10 exahertz (or >1019 Hz) and wavelengths less than 10 picometers (10.sup.−11 m). Typically, the gamma radiation is employed as irradiation to sterilise the composition or dressing.

[0087] In one embodiment the gamma radiation sterilises the composition or dressing. In one embodiment the gamma radiation is bacteriostatic. In one embodiment the gamma radiation is fungistatic. In one embodiment the gamma radiation reduces or eliminates the bioburden of the composition or dressing.

[0088] In one embodiment the gamma irradiation is cobalt 60 irradiation.

[0089] In one embodiment the gamma radiation is irradiation at approximately 20-50 kGy, such as approximately 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48 or 49 kGy, for example approximately 25-44.5 kGy or 25 kGy or more.

[0090] In one embodiment there is provided a composition comprising or consisting approximately: 85-95% purified water, 5-10% solvent, 0.5-1.0% thickener, 0.5-2.0% preservative, 0.1-1.0% mineral complex conditioning agent wherein each % means % w/w of the total composition.

[0091] In one embodiment there is provided a composition consisting essentially of 89.45% purified water, 8% propanediol, 0.8% sodium acryloyldimethyltaurate/VP crosspolymer, 1% phenoxyethanol and caprylyl glycol and chlorphenesin, 0.25% mineral complex and 0.5% (PHMB) polyaminopropyl biguanide (20% solution). In one embodiment the viscosity of the composition is approximately in the range 200 to 6000 cP.

[0092] In one embodiment there is provided a composition consisting essentially of 8% propanediol, 0.8% sodium acryloyldimethyltaurate/VP crosspolymer, 1% phenoxyethanol and caprylyl glycol and chlorphenesin, 0.25% mineral complex and 0.1% (PHMB) polyaminopropyl biguanide (20% solution) and purified water to make to 100%, such as approximately 89.85% purified water. In one embodiment the viscosity of the composition is approximately in the range 200 to 6000 cP.

[0093] The high water content of the composition enables it to absorb heat from the skin. Whilst not wishing to be bound by theory, the present inventors believe that this may help to reduce the development of burn by reducing the layers of skin cells permeated by the heat associated with burns.

[0094] In one embodiment the composition has a specific gravity of approximately 1.000+/−0.05 at 25° C.

[0095] In one embodiment the composition has a pH of approximately 5.5-7.5 at 25° C., such as approximately 5.0, 5.5, 6.5, 6.5 or 7.0, for example approximately 5.0-7.0.

[0096] In one embodiment the composition has a pH of approximately 4.0-6.5 at 25° C., such as approximately 4.5, 5.0, 5.5 or 6.0, for example approximately 4.0-6.5.

[0097] In one embodiment the topical composition is a fluid.

[0098] Fluid as employed herein means a low viscosity topical composition for application to unbroken skin. By contrast, creams and gels, including hydrogels, have a higher viscosity.

[0099] Advantageously, a lower viscosity means that the fluid is more easily absorbed by the skin and is easier to spread on the skin because it is less likely to drag the skin surface. This can be particularly useful where the patient is suffering pain or loss of skin integrity at the treatment site.

[0100] In one embodiment the composition is cooling.

[0101] In one embodiment the composition relieves pain.

[0102] In one embodiment the composition hydrates the skin.

[0103] A critical aspect of the present disclosure is the absorption of heat from the skin by the composition.

[0104] Thus, a critical aspect of the present disclosure is the reduction of the loss of skin fluid/moisture and structure by the composition.

[0105] In one embodiment there is provided a composition according to the disclosure for use as a medicament.

[0106] In one embodiment there is provided a composition according to the disclosure for use in the treatment or prophylaxis of burns. In one embodiment the burn is a thermal burn. In one embodiment the burn is a radiation burn. In one embodiment the burn is a chemical burn.

[0107] In one embodiment treatment with the composition relieves pain.

[0108] In one embodiment treatment with the composition reduces burning.

[0109] In one embodiment treatment with the composition reduces itching.

[0110] In one embodiment the composition is antimicrobial. In one embodiment the composition is antibacterial. In one embodiment the composition is antifungal.

[0111] As employed herein antimicrobial means that the composition is microbistatic or microbicidal. That is, it hinders the growth of, or kills microbes, including bacteria, fungi, viruses, protozoa, algae, amoebae and slime molds within the composition.

[0112] In one embodiment the composition reduces the depth of a burn.

[0113] In one embodiment the composition accelerates healing of the burn.

[0114] In one embodiment the composition reduces tissue necrosis.

[0115] In one embodiment the composition has substantially no oral toxicity.

[0116] In one embodiment there is provided a composition comprising water and one or more ingredients from the list consisting of: propanediol, sodium acryloyldimethyltaurate/VP crosspolymer, phenoxyethanol and caprylyl glycol and chlorphenesin, mineral complex and (PHMB) polyaminopropyl biguanide. Optionally the composition has a viscosity in the range 200-6000 cP. Optionally the viscosity of the composition is measured following exposure to gamma radiation.

[0117] Mineral Complex Conditioning Agent

[0118] Conditioning agents may have beneficial properties for wound healing. Without wishing to be bound by theory, it is believed that, following a burn injury, the body withdraws minerals from the skin it considers to be lost (that is, skin that will become necrotic). By replacing those minerals, in a bioavailable form, externally, it may be possible to save more of the skin from becoming necrotic and hence lost, thus requiring grafting therapy, or developing scarring.

[0119] Thus, in one embodiment the conditioning agent is a mineral complex conditioning agent. In one embodiment the mineral complex comprises bioavailable minerals, such as ion, free ions, elemental, or bound minerals, for example free ions.

[0120] In one embodiment the mineral complex comprises magnesium, potassium, sodium, boron, calcium and optionally one or more from the group consisting of: copper, nickel, silicon, zinc, aluminium, arsenic, barium, cadmium, cobalt, chromium, iron, mercury, manganese, lead, antimony, selenium, tin, strontium, titanium and vanadium.

[0121] In one embodiment the mineral complex is sea water extract. As employed herein sea water extract is the INCI name.

[0122] As employed herein sea water extract may be harvested from a deep sea source. Typically, the sea water extract is a concentrated solution of deep sea water minerals wherein the amount of sodium and/or chlorine has been reduced and/or substantially eliminated.

[0123] In one embodiment the sea water extract is dead sea salt, Cornish sea salt, Maldon sea salt, Himalayan sea salt and the like.

[0124] In one embodiment the mineral complex is Epsom salts.

[0125] In one embodiment the sea water extract is the INCI and IUPAC name.

[0126] In one embodiment the sea water extract is Deep Sea Water provided by Morechem. In one embodiment the sea water extract is Eau de Source Marine SC, Ocaline or Ocaline XP provided by Soliance (Givaudan) or the like.

[0127] In one embodiment the mineral complex conditioning agent is added to the composition in liquid form, such as a concentrate of sea water.

[0128] In one embodiment the mineral complex conditioning agent is added to the composition in dried form. For example, as dried, concentrate of sea water.

[0129] In one embodiment the mineral complex does not comprise bound minerals such a magnesium sulphate/oxide/citrate.

[0130] In one embodiment the mineral complex comprises free magnesium, such a Mg.sup.2+ ions. In one embodiment the major component of the mineral complex is magnesium.

[0131] In one embodiment the mineral complex comprises potassium, such as free potassium, such as K.sup.+ ions.

[0132] In one embodiment the mineral complex comprises sodium, such as free sodium, such as Na.sup.+ ions.

[0133] In one embodiment the mineral complex comprises boron, such as free boron, such as boron anions or boron cations.

[0134] In one embodiment the mineral complex comprises calcium, for example free calcium, such as Ca.sup.2+ ions.

[0135] In one embodiment the mineral complex provides bioavailable minerals, such as magnesium.

[0136] In one embodiment the mineral complex has substantially no chloride or chlorine.

[0137] In one embodiment the sea water extract is Oriel sea water extract (orielmarineextracts.com) provided by Oriel Sea Salt Co.

[0138] In one embodiment the sea water extract has a pH of approximately 7 to 8, such as approximately 7.4.

[0139] In one embodiment the sea water extract has a density of approximately 40%.

[0140] Table 1 shows the components of sea water.

TABLE-US-00001 TABLE 1 Atomic Element weight p m Hydrogen H2O 1.0079 110,000 Oxygen H2O 15.99 883,000 Sodium NaCl 22.989 10,800 Chlorine NaCl 35.453 19,400 Magnesium Mg 24.312 1,290 Sulfur S 32.064 904 PotassiumK 39.102 392 Calcium Ca 10.080 411 Bromine Br 79.909 67.3 Helium He 4.0026 0.0000072 Lithium Li 6. 0.170 Beryllium Be 9.0133 0.0000006 Boron B 10.811 4.450 Carbon C 12.011 28.0 Nitrogen ion 14.007 15.5 Flourine F 18.998 13 Neon Ne 20.183 0.00012 Aluminum Al 26.982 0.001 Silicon Si 28.086 2.9 Phosphorus P 3 .974 0.088 Argon Ar 39.948 0.450 Scandium Se 44.956 <0.000004 Titanium Ti 47. 00 0.001 Vanadium V 50.942 0.0019 Chromium Cr 51.996 0.0002 Magnesium Mg 54. 38 0.0004 Ferrum (Iron) Fe 55.847 0.0034 Cobalt Co 58.933 0.00039 Nickel Ni 58.710 0.00 6 Copper Cu 63.54 0.0009 Zinc Zn 65.37 0.005 Gallium Ga 69.72 0.0000 Germanium Ge 72.59 0.00006 Arsenic As 74.922 0.0026 Selenium Se 78.96 0.0009 Krypton Kr 83. 0.00021 Rubidium Rb 85.47 0.120 Strontium Sr 87.62 8.1 Yttrium Y 88.904 0.000013 Zirconium Zr 91.22 0.000026 Niobium Nb 92.906 0.000015 Molybdenum Mo 0. 594 0.1 Ruthenium Ru 102.905 0.0000007 Rhodium Rh 106.4 — Palladium Pd 170.870 — Argentum 107.870 0.00028 (silver) Ag Cadmium Cd 112.4 0.00011 Indium In 114.83 — Stannum (tin) Sn 118. 0.00081 Antimony Sb 121.75 0.00033 Tellurium Te 127.6 — Iodine I 166.904 0.064 Xenon Xe 131.30 0.000047 Cesium Cs 132.905 0.000 Barium Ba 137.34 0.021 Lanthanum La 138.91 0.0000029 Cesium Cs 140. 2 0.0000012 Praseodymium 140.907 0.00000064 Pr Neodymium Nd 144.24 0.0000028 Samarium Sm 150. 5 0.00000045 151.96 0.00000 3 Godolinium Gd 157.25 0.00000007 Terbium Tb 158.924 0.00000014 Dysprosium Dy 162.50 0.00000091 Holmium Ho 164.930 0.00000022 Erbium Er 167.26 0.00000087 Thulium Tm 168.934 0.00000017 Ytterbium Yb 173.04 0.00000082 Lanthanum La 174.97 0.00000015 Hafnium Hf 178.49 <0.000008 Tantalum Ta 180.948 <0.0000025 Tungsten W 183.85 <0.000001 Rhenium Re 186.2 0.0000084 Osmium Os 190.2 — Iridium Ir 192.2 — Platinum Pt 195.09 — Aurum (gold) Au 196.967 0.000011 Mercury Hg 200.59 0.00015 Thallium Tl 204.37 — Lead Pb 207.19 0.00003 Bismuth Bi 208.980 0.00002 Thorium Th 232.04 0.0000004 Uranium U 238.03 0.00 3 Plutonium Pu (244) — indicates data missing or illegible when filed

[0141] In one embodiment the mineral complex comprises approximately: 66% magnesium, 23.8% potassium, 9.8% sodium, 0.002% boron, 0.0006% calcium, 0.00002% copper, 0.000012% nickel, 0.0000087% silicon and 0.000001% zinc. Wherein approximately is defined to be +/−15%. In one embodiment the mineral complex further comprises trace elements. In one embodiment the trace elements include one or more from the group: aluminium, arsenic, barium, cadmium, cobalt, chromium, iron, mercury, manganese, lead, antimony, selenium, tin, strontium, titanium and vanadium. In one embodiment the trace elements may be any element selected from Table 1.

[0142] In one embodiment the mineral complex comprises one or more minerals according to Table 1.

[0143] Dressing Material

[0144] A burn dressing in accordance with the present disclosure is formed by impregnating a suitable dressing material with the composition of the disclosure.

[0145] Dressing material as employed herein means a fabric carrier capable of holding a chosen volume of composition. Preferably the dressing material is a non-woven synthetic material that will hold a substantial quantity of the composition to apply an effective amount of the composition to a burn. The dressing material must be capable of being sterilised, typically by irradiation, such as gamma irradiation and non-irritating to burned skin.

[0146] In one embodiment there is provided a burn dressing comprising a topical composition according to the disclosure and a dressing material.

[0147] In one embodiment the dressing material comprises thermal bonded, non-woven material.

[0148] In one embodiment the dressing material is polyester, PET (polyethylene terephthalate) or the like, such as medical grade non-woven 100% polyester fabric, for example polypropylene or rayon.

[0149] Thermal bonded as employed herein means a fabric wherein heat energy is used to stimulate an adhesive, which in turn flows to thermoplastic fibre juncture and interlocks the fibres upon cooling.

[0150] Non-woven as employed herein refers to sheet or web structures bonded together by entangling fibre or filaments (and by perforating films) mechanically, thermally or chemically. They are flat, porous sheets that are made directly from separate fibres or from molten plastic or plastic film.

[0151] In one embodiment the dressing material comprises super absorbent material, such as super absorbent fibre.

[0152] Super absorbent materials have an absorbent capacity of several times their weight. Super absorbent fibres are fibrous form of super absorbent material which can be incorporated into woven or non-woven materials.

[0153] In one embodiment the dressing material comprises polypropylene fibre and rayon fibre.

[0154] In one embodiment the dressing material comprises super absorbent fibre, polypropylene fibre and rayon fibre.

[0155] In one embodiment the dressing material comprises approximately 10-40% super absorbent fibre, such as approximately 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38 or 39% super absorbent fibre, for example approximately 20% super absorbent fibre.

[0156] In one embodiment the dressing material has one of more of the properties selected from the group consisting: a weight of approximately 50 gsm, a thickness of approximately 0.63 mm, a tensile strength of approximately 4.2 N or 24.3 N, an absorbent capacity of approximately 22.7 g/g and an absorbent volume of approximately >1150 gsm.

[0157] In one embodiment the dressing material is type 2741 fabric as provided by Technical Absorbents.

[0158] In one embodiment the dressing has a width of approximately 5 cm to 50 cm and a length of approximately 5 cm to 50 cm. Such as approximately 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, or 49 cm width and/or length.

[0159] In one embodiment the dressing material holds approximately 15 to 30 grams of composition per gram, such as approximately 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28 or 29 grams of composition per gram, for example approximately 22.7 g/g.

[0160] In one embodiment the dressing material holds approximately 1000 to 2000 g of composition per square metre of dressing material, such as approximately 1100, 1200, 1300, 1400, 1500, 1600, 1700, 1800 or 1900 g of composition per square metre of dressing material. For example, approximately 1674 g of composition per square metre of dressing material.

[0161] In one embodiment the dressing is of shape and dimension suitable for application to the face.

[0162] In such embodiment the dressing may have slots or holes for the eyes and/or nose and/or mouth.

[0163] In one embodiment the dressing material has pockets in which the composition may be placed. For examples, see EP0521143 which is incorporated herein by reference.

[0164] Sterilisation

[0165] In one embodiment the composition or dressing is sterilised, for example by heat (such as by steam or dry heat), irradiation (such as electron beam or gamma radiation), gas (such as ethylene oxide or formaldehyde) or low temperature oxidative sterilisation (such as vaporised hydrogen peroxide, hydrogen peroxide/gas plasma).

[0166] In one embodiment the composition or dressing is sterilised by gamma irradiation.

[0167] In one embodiment the gamma irradiation is cobalt 60 or caesium 137 radiation, particularly cobalt 60 radiation.

[0168] In one embodiment the composition or dressing is irradiated to meet 10E6 sterility assurance level (SAL).

[0169] In one embodiment the sterilisation method is AAMI 11137-2 compliant.

[0170] Advantageously, compositions and dressing that have been sterilised employing the method have substantially zero bioburden. That is, they have zero CFUs. Such as no microbe that can replicate or grow.

[0171] Packaging

[0172] In one embodiment the burn dressing as disclosed herein is packaged into a storage pouch. Advantageously, the storage pouch permits the dressing to remain sterile and be easily transported, for example is a first aid kit or medical kit, such as for use by a paramedic.

[0173] Typically, the storage pouch has a three-layer construction of a layer of polyester having a layer of aluminium thereon and a layer of, for example, Scotchpak® heat sealable polyester film thereof. The three layers are adhered with adhesive.

[0174] The compositions, dressings and methods of the present disclosure when employed help maintain skin integrity, minimise the deleterious effects of burns and reduce opportunistic infections that may occur when skin is damaged.

[0175] The maintenance of moisture around the burn may also minimise scarring and prevent reduced flexibility in the area of skin damage. This is advantageous because it may reduce pain associated with scar tissue and avoids skin thickening and reduced skin elasticity which, in skin folds, can be problematic.

[0176] It is desirable to avoid skin toughness that can arise following damage to the skin because toughened skin is prone to flaking and cracking which in turn can lead to inflammation and infection.

[0177] In one embodiment damaged cells treated with the topical composition or dressing recover viability more quickly than untreated cells. In one embodiment cell viability is restored more quickly in cells treated with the topical composition or dressing.

[0178] In one embodiment there is provided a burn dressing for use in the treatment or prophylaxis of burns. Typically, the burn dressing comprises a composition as disclosed herein absorbed and carried on or in a dressing material as described herein.

[0179] Ideally the composition or dressing as described herein is applied to a burn as soon as possible following the burn. Preferably the composition or dressing is applied immediately, such as within a minute of the burn. The composition or dressing may be applied within a few hours of the burn injury.

[0180] In some situations, the composition or dressing may be applied following treatment by a medical professional. That is, the composition or dressing may be employed other than as a first aid treatment. For example, the composition or dressing may be employed for prolonged use, for example, to keep a burn wound sterile and/or hydrated. Such use of the composition or dressing supports the skin cells by providing external bioavailable minerals which, it is thought, supports the increased metabolism of the cells.

[0181] In one embodiment the composition or dressing is applied once, twice, three or four a day.

[0182] In one embodiment the composition or dressing is applied to skin, such as the area of the burn, and left for approximately 10 minutes to 36 hours, for example approximately 20, 30, 40 or 50 minutes or approximately 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34 or 35 hours. In one embodiment the composition or dressing is applied to a burn for up to approximately 24 hours.

[0183] In one embodiment there is provided a composition or dressing for use in treatment of a burn wherein the treatment is prolonged treatment.

[0184] In one embodiment there is provided a method of prophylaxis or treatment wherein the composition or dressing is applied to a burn for approximately 24 hours. For example approximately 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35 or 36 hours or more.

[0185] In one embodiment treatment with the composition or dressing continues for about 2 to 10 weeks following each burn injury, such as 3, 4, 5, 6, 7, 8, or 9 weeks following burn injury.

[0186] Typically, the composition or dressing is changed daily and a new composition or dressing according to the disclosure applied to the burn injury.

[0187] In one embodiment the composition or dressing provide bioavailable minerals to the skin. In one embodiment the minerals include magnesium. It is believed that bioavailable magnesium may help prevent magnesium depletion which is known to be a complicating factor in burn injuries. In one embodiment the minerals include calcium.

[0188] In one embodiment the composition or dressing promote faster healing of the burn wound. In one embodiment use of the composition or dressing results in reduced scarring.

[0189] In one embodiment there is provided a composition or dressing as disclosed herein for use in reducing scarring.

[0190] Thus, there is provided a composition or dressing for direct application to a burn wound. The dressing can be employed to cover the entire burn. Debridement of the burn is not necessary prior to application of the composition or dressing. The composition rapidly penetrates clothing and wets, cools and soothes a burn. The burn is wet, cooled and soothed, not only on the surface but beneath the surface, thereby reducing progression of the burn. The burn dressing cools by heat transference and helps create an isothermic environment. Additionally, the composition or burn dressing helps reduce contamination of the burn by covering the burn and blocking air-borne microbes. Clothing and skin do not adhere to the burn dressing when it is removed, thereby limiting pain and skin damage when the dressing is removed.

[0191] The composition and dressing are non-toxic, water-soluble and retain properties after extended storage. Advantageously, the composition and dressing are easy to use.

[0192] In the context of this specification “comprising” is to be interpreted as “including”.

[0193] Approximately, as used herein, means +/−10%.

[0194] Aspects of the invention comprising certain elements are also intended to extend to alternative embodiments “consisting” or “consisting essentially” of the relevant elements.

[0195] Where technically appropriate, embodiments of the invention may be combined.

[0196] Embodiments are described herein as comprising certain features/elements. The disclosure also extends to separate embodiments consisting or consisting essentially of said features/elements.

[0197] Technical references such as patents and applications are incorporated herein by reference.

[0198] Any embodiments specifically and explicitly recited herein may form the basis of a disclaimer either alone or in combination with one or more further embodiments.

[0199] The present invention is further described by way of illustration only in the following examples:

EXAMPLES

Example 1

[0200] Following several failed attempts to formulate a composition with suitable viscosity and other properties to function as a burn treatment, the Inventors obtained stable compositions which were sent for testing to assess stability under gamma radiation.

[0201] OVERVIEW: To incorporate: Polyaminopropyl biguanide (INCI name);

[0202] Chemical name: Polyhexamethylene Biguanide Hydrochloride (PHMB) and later Oriel sea mineral complex into a gel formula that can withstand the impact of gamma radiation sterilisation.

[0203] 5 rounds of formulas were sent out for gamma radiations as outlined below.

[0204] Round 1 Summary: Started with our current BD (Burn Dressing) Gel with hyaluronic acid (HA) formula to which various ingredients were added.

[0205] The table below shows the key ingredients added to BD gel w/HA formula to determine their impact on gamma radiation resistance (Experiments A through L). Experiment L containing Carbopol and water only, shows that PHMB @1% (20% solution) is incompatible with Carbopol (thickening agent). The gel curdles. Carbopol is the thickening agent used in BD Gel with HA, the only experiment in round 1 to which PHMB was added.

TABLE-US-00002 TABLE 2 Discolour- Key ation ingredients following (Round 1) Experi- BD and Ingredient radiation ment HA plus: INCl/Name Function (1-10) A Glycerin Glycerin Humectant 1.2 B Propylene 1,2-Propanediol Humectant 1.0 glycol C Tinoguard HS Sodium UV absorber 4.0 (BASF) Benzo Butylphenol Sulfonate D Ciba Sodium UV absorber 1.5 benzotriaxolyl Liquid (BASF) B tylphenol Sulfonate E Ti Penacry- UV absorber 2.5 (slightly TT (BASF) Tetra-di- -butyl hazy) F PHMB Poly- Preservative 3 aminoproyl (off white, b hazy) G A, B, C, D, E See above See above 3.5 (off white, hazy) H A, B, C, D, E, F See above See above 1.5 I microsilver Not s  radiation - too dark J Control - 1.2 additional ingredients K Control - just Carbopol and water, not irradiated L Control - just 6.0 Carbopol water and lamine indicates data missing or illegible when filed

[0206] After gamma results: Discolouration was measured on a scale of 1 (no discolouration) to 10 (intense discolouration) UV absorbers showed some discolouration; propylene glycol showed little or no change following gamma radiation.

[0207] Round 2 Summary: Since PHMB was incompatible with Carbopol, new formulas containing various other thickeners were tried. Since propylene glycol, a humectant helped, another humectant (propanediol) was tried. For all the experimental batches made, only the stable formulas were sent out for gamma radiation. Only some of round 2 formulas contain PHMB (20% solution) @0.2%.

[0208] The base gel employed in experiments is water plus thickener (Natrosol or Laponite for example).

TABLE-US-00003 TABLE 3 Key Ingredients Ingredient INCl/ (Round 2) Chemical Name Function Natrosol 250 HHX Pha Thicking agent hydroxy Propylene glycol Propylene glycol ec Laponite XL 21 Sodium Magnesium Thickening agent Fl silicate Propanediol Propanediol Solvent (1,3-Prop ) Xantham gum Xantham gum Thicking agent Carrageenan Carrageenan Thicking agent Aculyn 21 Acrylates/Stea -20 Thicking agent Methacrylate Copolymer Agar Powder Agar Thicking agent Poloxamer 188 poloxamer 188 Surfacant/Thickening agent Glycerin Glycerin Hu ectant indicates data missing or illegible when filed

[0209] Formulas with xanthan gum carrageenan, aculyn 46 N, poloxamer 188 and agar were unstable/thinned out or discoloured (thickening agents not compatible with PHMB) and therefore gels were not sent out for gamma radiation.

[0210] After Gamma Radiation Results:

[0211] Formulas containing Natrosol 250 HEX completely loss viscosity and became “water thin” but gel was not discoloured. Formulas with propylene glycol were clear but also had a pinkish hue. Formulas with propanediol remained clear; those with glycerin acquired a yellowish hue.

[0212] Round 3 Summary: For round 3 experiments, 2 new thickening agents (Sodium Carboxymethyl Cellose and Aristoflex AVS) were tested.

TABLE-US-00004 TABLE 4 Key Ingredients (Round 3) Water Ingredient INCl/ Experiment plus: Chemical Name Function A Sodium Sodium Thickening Carboxymethyl Carboxymethyl agent Cellose and PHMB Cellose B A plus propanediol C B plus Mikrokill and disodium EDTA D A plus disodium EDTA, propylene glycol, Mikrokill E Disodium EDTA, propanediol Mikrokill, PHMB, Carbopol 980, trolamine F Disodium EDTA, Sodium Thicking propanediol, Acryloyldimethyltau- agent Mikrokill, PHMB, rate/VP Crosspolymer Aristoflex AVS G Disodium EDTA propanediol, Mikrokill, PHMB, Natrosol HHX, Carbopol 980, trolamine

[0213] All the formulas in round 3 contained PHMB 0.2% (20% solution). Only stable formulas were sent out for gamma radiation.

[0214] After Gamma Radiation Results:

[0215] Formulas containing sodium carboxymethyl cellose became watery.

[0216] Although the combination of Carbopol and Natrosol 250 HHX showed some promising results (Exp. G), the best result was EXP.F which contained a combination of propanediol and Aristoflex AVS.

[0217] Round 4 Summary: In round 4 experiments, Oriel sea mineral extract skin conditioner was introduced into the formulas. This ingredient lowers the viscosity of the gel. As in Round 3 experiments, PHMB was still used @ 0.2% (20% solution). Experiment F, (Round 3) having the best results from round 3 was the starting point. The level of Aristoflex AVS (thickening agent) was varied to compensate for the viscosity reducing effect of the Oriel sea mineral extract. The levels of propanediol were also varied from 5% to 12% to see what if any effect that had on the gamma radiation results as well on overall product appearance.

TABLE-US-00005 TABLE 5 Key Ingredients Ingredient INC /Chemical (Round 4) Name Function Oriel Sea Mineral Sea water Skin conditioning Extract extract agent Aristoflex AVS Sodium Thicking agent Acryloyldimethyl rate/VP Cross polymer Propylene glycol propylene glycol Solvent (1,2-propanediol) Propanediol Propanediol Solvent (1,3-Propanediol) Carbopol 980 Cad omer Thicking agent natrosol 250 HHX Pharm hydroxyethylecelluose Thicking agent indicates data missing or illegible when filed

[0218] Only stable formulas were sent out for gamma radiation.

[0219] After Gamma Results:

[0220] All the experiments containing a combination of Aristoflex AVS, PHMB and propanediol showed good results regardless of the level of propanediol. Compositions with propanediol and Carbopol but without PHMB had good results.

[0221] Compositions with propanediol/Carbopol/PHMB combination showed a significant decrease in viscosity.

[0222] Round 5 Summary: Round 5 experiments involved: (a) optimising the viscosity of the product to work more efficiently with the new absorbent material. (b) Increasing the level of PHMP from 0.2% to 0.5% (20% solution). (c) Making formulas for preservative challenge without the main preservative Mikrokill®COS but with PHMB along with various levels of propanediol (which has preservative properties). Note: Final formula contains Mikrokill®COS. (d) Optimising the manufacturing process.

TABLE-US-00006 TABLE 6 Key Ingredients Ingredient INCl/Chemical (Round 5) Name Function Oriel Sea Mineral Sea water extract Skin conditioning Extract agent Aristoflex AVS Sodium Thicking agent Acryloyldimethyl rate/VP Crosspolymer Propylene glycol Propylene glycol Solvent (1,2-Propanediol) Propanediol Propanediol Solvent (1,3-Propanediol) indicates data missing or illegible when filed

[0223] In round 5, the final formula was determined from a selection of which were sent out for gamma radiation with acceptable results. All the formulas are similar except for their levels of Aristoflex AVS (thickening agent) varying from 1.0%, 0.9% and 0.8% respectively. A decision was made to go with a formula with 0.8% Aristoflex AVS (final formula), the least viscous formula.

[0224] In order to test physical integrity of the composition following gamma radiation, the viscosity at room temperature and 40° C. can be tested and compared to a control which was not irradiated.

Example 2

[0225] Wound healing progresses via three overlapping phases: inflammation, granulation and tissue remodelling. After cutaneous injury, a blood clot forms, and inflammatory cells infiltrate the wound, secreting cytokines and growth factors to promote the inflammation phase. During the granulation phase, fibroblasts and other cells differentiate into myofibroblasts, which deposit extracellular matrix (ECM) proteins. Simultaneously, angiogenesis occurs, and keratinocytes proliferate and migrate to close the wound. In the final tissue-remodelling phase, apoptosis eliminates myofibroblasts and extraneous blood vessels, and the ECM is remodelled to resemble the original tissue. Dysregulation of this last tissue remodelling phase leads to fibrosis.

[0226] In order to monitor this cytotoxicity, behaviour, impact and biofunctionality of the composition in (1) Human Vascular Endothelial Cells, (2) Human Dermal Fibroblasts and (3) Human Dermal Keratinocytes we employed an electrical-impedance based technique that monitors and quantifies in real-time the behaviour of cells, which is also amenable to high throughput. Giaever and Keese first described a technique for measuring fluctuations in impedance based on the principle of population cell growth on a specialized electrode surface. The xCELLigence instrument, established and optimised in the laboratory of Dr Ronan Murphy (Dublin City University), utilises a similar technique to measure changes in electrical impedance. Through preliminary studies and data from working with the ‘mineral-complex’ ingredient, we have determined protocols and conditions that are optimal for cell functionality and activation in all three cell types. For this we used a 2.5D model on e-plates. Briefly, as cells attach and spread in a culture dish covered with a gold microelectrode array that covers approximately 80% of the area on the bottom of a well. As cells attach and spread on the electrode surface, it leads to an increase in electrical impedance. The impedance is displayed as a dimensionless parameter termed cell-index, which is directly proportional to the total area of tissue-culture well that is covered by cells. Hence, the cell-index can be used to monitor many critical stages of cell behaviour such as wound healing: cell adhesion, spreading, morphological changes, detachment, proliferation, migration, apoptosis and cell density.

[0227] The standard wound healing assay was utilised in this study based on changes in electrical impedance at the electrode/cell interphase, as a population of cells migrates an advanced double chamber apparatus know as a CIM plate. Cell migration, fate, function and behaviour lead to large changes in impedance. These changes directly correlate with the wound healing capacity of the three cell types, i.e., migration and tissue/ECM remodeling by cells lead to large changes in cell impedance and vice versa. This advanced wound-healing assay involved a two-chamber system (xCELLigence CIM (cell invasion and migration) plate) to monitor and measure transmigration as well as initial surface layer disruption. This technique provides a two-fold advantage over existing methods of measuring invasion, such as Boyden chamber and matrigel assays: firstly, the Cell-Extra Cellular Matrix interactions and remodeling more closely mimics the in vivo process, and secondly, the data was obtained in real-time and is more easily quantifiable, as opposed to end-point analysis for other methods.

[0228] Dermal fibroblasts are cells that lay within the dermis layer of skin and are responsible for generating connective tissue and allowing the skin to recover from injury. Dermal fibroblasts generate and maintain the connective tissue which unites separate cell layers, particularly via the rough endoplasmic reticulum. Crucially, it is these dermal fibroblasts that produce the protein molecules, including laminin and fibronectin, which comprise the extracellular matrix (ECM). Hence, by creating the ECM between the dermis and epidermis, fibroblasts facilitate the epithelial cells of the epidermis to affix the matrix, thereby allowing the epidermal cells to effectively join together to form the top layer of the skin.

[0229] In our experiments, dermal fibroblast cells were grown in culture, starving them of magnesium for 24 hours before treating them to (NB105-142) & appropriate controls. Cells were seeded onto 0.32 cm.sup.2 wells of the xCELLigence real-time monitoring system, upon which, a minimal layer of ECM had been permitted to form. Cells were then allowed to adhere to the electrode surface and migrate accordingly. Results are presented In FIGS. 1 and 2.

Example 3

[0230] We employed the Wound Healing RT2 Profiler PCR Array to assess the effect of the composition on gene expression during the process outlined in Example 2. This time both fibroblast monoculture (Example 3a) and our established human LabSkin model (see Duffy Et al, 2017, Cosmetics, 4, 44) was used (Example 3b).

[0231] This array contains genes important for each of the three phases of wound healing, including ECM remodeling factors, inflammatory cytokines and chemokines, as well as growth factors and major signaling molecules. Using real-time PCR, you can easily and reliably analyse the expression of a focused panel of genes involved in wound healing, tissue injury and repair with this array. The RT2 Profiler PCR Array System is the most reliable and accurate tool for analysing the expression of a focused panel of genes using SYBR Green-based real-time PCR. It brings together the quantitative performance of real-time PCR and the multiple gene profiling capability of microarrays. Each PCR Array profiles the expression of 84 genes relevant to a specific pathway or disease state-in this case Wound Healing. Expression levels are measured by gene-specific RT2 qPCR Primer Assays optimized for simultaneous use in the PCR Array System. RT2 qPCR Primer Assays are key components in the PCR Array System. Each qPCR assay on the array is uniquely designed for use in SYBR Green real-time PCR analysis. The assay design criteria ensure that each qPCR reaction will generate single, gene-specific amplicons and prevent the co-amplification of non-specific products. The qPCR Assays used in PCR Arrays are optimised to work under standard conditions enabling a large number of genes to be assayed simultaneously. This system is specifically designed to meet the unique challenges of profiling pathway-focused sets of genes using real-time PCR. Simultaneous gene expression analyses require similar qPCR efficiencies for accurate comparison among genes. RT.sup.2 qPCR Primer Assays are designed with an amplicon size ranging from 100 to 250 bp and with PCR efficiencies uniformly greater than 90%. Overall, more than 10 thermodynamic criteria are included in the design of each RT.sup.2 qPCR Primer Assay to ensure the most reliable and accurate results for pathway-based gene expression analysis in the PCR Array System. The array layout is shown in Table 7 below.

TABLE-US-00007 TABLE 7  Ref Seq Number  Description A01 NM_  ACT 2 , alpha 2, A02 NM_  ACTC  , alpha, , 1 A03 NM_  AN PT1 1 A04 NM_  CCL2 C (CC ) 2 A05 NM_  CCL7 C (CC ) 7 A06 NM_ A07 NM_ 004 CD Cad 1, 1, ( ) A08 NM_ 021 COL Colla , type XIV, alpha 1 A09 NM_  COLLA1 Colla , type 1, alpha 1 A10 NM_  COLLA2 Colla , type 1, alpha 2 A11 NM_  COL Coll, type III,  1 A12 NM_  COL Coll, type IV, alpha B01 NM_  COL Coll, type IV, alpha ( ) B02 NM_  COL5A1 Coll , type V, alpha 1 B03 NM_  COL5A2 Coll , type V, alpha 2 B04 NM_  COL5A3 Coll , type V, alpha 3 B05 NM_  ( ) B06 NM_ B07 NM_ CT F B08 NM_ CTNN , B09 NM_  G B10 NM_  K B11 NM_  L2 B12 NM_ (CXC) 1 , ) C01 NM_ CXC  Cl (C X C)  11 C02 NM_ CXC 2 C (C X C)  2 C03 NM_ 2994 CXC15 C(C X C)  3 C04 NM_E C05 NM_ C06 NM_ C07 NM_ ( ) C08 NM_ T A alpha C09 NM_ 1 C10 NM_ TGF2 2 ( ) C11 NM_ P  7 C12 NM_ EGF D01 NM_ HGF( ) D02 NM_ D03 NM_  C) D04 NM_  10 D05 NM_  1, beta D06 NM_  2 D07 NM_  4 D08 NM_ 6 ( ,  2) D09 NM_ 6 ( , M ) D10 NM_ , 1 D11 NM_ , 2 ( , 2 VLA 2) D12 NM_ , 3 ( , , 3 3 ) E01 NM_ , 4 ( , , 4 4 ) E02 NM_ , ( , , alpha ) E03 NM_ , 6 E04 NM_ , (, , alpha , CD) E05 NM_ , 1 (, , beta , CD MD 2, M K12) E06 NM_ , 3 C E07 NM_ , E08 NM_ ,  6 E09 NM_  MAPK  M E10 NM_  MAPK M E11 NM_  (gly ) E12 NM_  MMP1 M 1 ( ) F01 NM_  MMP2 M 2 (, 72 kDa , 72 kDa type IV colla ) F02 NM_  MMP  M  7 ( , ) F03 NM_  MMP  M  9 ( , 92 kDa , 72 kDa type IV colla ) F04 NM_  Pla  growth factor pha poly F05 NM_  PLAT P F06 NM_ PLAU P F07 NM_  PLAUR P , F08 NM_  PL Pl F09 NM_  PTEN P F10 NM_  PT S2 2 ( ) F11 NM_  RAC  R  (, GTP Rac ) F12 NM_  RHOA , G01 NM_  SERPINE  rpi  p , activator type ) G02 NM_  STAT  3 (  response factor) G03 NM_ TA LN T G04 NM_  T FA T  growth factor, alpha G05 NM_ T FAB  T  growth factor, beta G06 NM_ T FABR  T  growth factor, beta receptor G07 NM_ TIMP  TIMP G08 NM_ TNF T G09 NM_ V FA Vascular  growth factor G10 NM_  VTN V G11 NM_  WR  WNT  pathway G12 NM_  WNT5A W  -type MM  family  member 5 indicates data missing or illegible when filed

Example 3a

[0232] In fibroblast monoculture, 2 genes were found to be upregulated and 22 were downregulated when treated with the composition versus the control (untreated cells). Results are shown in FIG. 3a and Table 8 below.

TABLE-US-00008 TABLE 8 Position Gene F d R Up-Regulated Genes COL 19.2929 PLO 5.579 Down-Regulated Genes A02 ACTC1 −11.0809 A0 CD40LG −14.8254 B10 CT K −4.2871 B CTSV −9.5798 C01 CXCL11 −4.5948 C04 EGF −33.8246 C06 13A1 −92.4 15 C07 F3 −7.3107 C09 FGF −27.4741 D05 B −4.084 D07 −15.8895 D10 IT A1 −5. 281 E05 IT B1 − .27 7 E07 IT B5 −4. E08 IT B6 −16.44 8 F09 PTEN −8.5742 F11 RACl −8. 39 F12 RBO − .021 02 STAT3 −4.5 48 10 VTN − .81 9 12 WNT5 −16.223 indicates data missing or illegible when filed

Example 3b

[0233] Development of In Vitro Human 3D Deep-Skin Technology & Application in Burn Research

[0234] A highly advanced 3D living skin equivalent model (developed by Dr Ronan Murphy's team at Dublin City University) is unique in providing unrivalled opportunities for non-animal testing and research. The fully differentiated epidermis is supported by a dermal component consisting of fibroblasts in a fibrin matrix. The model also allows micro-organisms to be grown on its surface, mimicking infection or the skin's natural microflora. This configuration ensures we can assess topical formulations with possibly the most comprehensive range of tests available in an in vitro model. A schematic of the system is shown in FIG. 4. Culture medium 10 sits below the skin 20 to provide nutrients for growth. The resulting skin is stratified as shown in the cross section 30.

[0235] Skin Model Burn Protocol

[0236] Custom 3.66 g brass weights were milled from brass stock with a surface contact area of 10 mm and a protrusion for handling with tweezers (see FIG. 5a). The weights were heated to 100° C. on a heating block (Stuart) and temperature checked using an IR thermometer.

[0237] Skin models were removed from the 6-well plate and placed onto a plastic surface in a laminar hood to avoid heat dissipation. Brass weights were removed from the heating block using tweezers and immediately placed on the centre of each 2.5 cm model for 10 seconds. After 10 seconds, the brass weight was removed and the appropriate treatment was applied.

[0238] Each treatment consisted of custom cut 2.5 cm gauze disks (Water-jel) soaked in different formulations.

[0239] Model skin turned white in the centre following removal of the weight. FIGS. 6a and 6b show photographs of six of the models (numbered 1-6) 24 hours after the burn infliction.

[0240] All models were biopsied using a 3 mm biopsy punch (Miltex) in the centre and at the burn boundary 24 and 48 hours after the burn was inflicted (see FIG. 5b), and conditioned media was sampled.

[0241] Genes associated with wound (burn) repair are:

[0242] Extracellular Matrix & Cell Adhesion:

[0243] ECM Components: COL14A1, COL1A1, COL1A2, COL3A1, COL4A1, COL4A3, COL5A1, COL5A2, COL5A3, VTN.

[0244] Remodelling Enzymes: CTSG, CTSK, CTSL2, F13A1, F3 (Tissue Factor), FGA (Fibrinogen), MMP1, MMP2, MMP7, MMP9, PLAT (tPA), PLAU (uPA), PLAUR (uPAR), PLG, SERPINE1 (PAI-1), TIMP1.

[0245] Cellular Adhesion: CDH1 (E-cadherin), ITGA1, ITGA2, ITGA3, ITGA4, ITGA5, ITGA6, ITGAV, ITGB1, ITGB3,

[0246] ITGB5, ITGB6.

[0247] Cytoskeleton: ACTA2 (a-SMA), ACTC1, RAC1, RHOA, TAGLN.

[0248] Inflammatory Cytokines & Chemokines:

[0249] CCL2 (MCP-1), CCL7 (MCP-3), CD40LG (TNFSF5), CXCL1, CXCL11 (ITAC/IP-9), CXCL2, CXCL5 (ENA-78/LIX), IFNG, IL10, IL1B, IL2, IL4, IL6.

[0250] Growth Factors:

[0251] ANGPT1, CSF2 (GM-CSF), CSF3 (GCSF), CTGF, EGF, FGF10, FGF2, FGF7, HBEGF (DTR),

[0252] HGF, IGF1, MIF, PDGFA, TGFA, TGFB1, TNF, VEGFA

[0253] Signal Transduction:

[0254] TGF: TGFB1, TGFBR3, STAT3.

[0255] WNT: CTNNB1, WISP1, WNT5A.

[0256] Phosphorylation: MAPK1 (ERK2), MAPK3 (ERK1), PTEN.

[0257] Receptors: EGFR, IL6ST (GP130).

[0258] Other: PTGS2.

[0259] Firstly, RT2 qPCR was first employed to compared burned skin to healthy skin to obtain a baseline. FIG. 8a shows up- and down-regulated genes in 3D skin models in response to thermal burn injury (no treatment) vs healthy skin. Total RNA from 3D skin models was characterised, and the relative expression levels for each gene in the two samples (burn vs healthy skin) plotted against each other in the Scatter Plot. Table 10 shows 22 genes that are upregulated in burned skin relative to unburned skin. Table 11 shows 49 genes that are down regulated in thermally injured (burned) skin relative to unburned skin.

TABLE-US-00009 TABLE 10 Position Gene Fold Change C03 CXC 5 6942.47 06 ITGB3 785.57 C06 F13A1 620.97 B05 CSF2 503.94 08 TNF 156.36 D06 IL2 112.45 C08 FGA 105.14 C01 CXCL11 45.2 D08 IL6 36.04 F10 PT 2 12.85 A 4 CCL2 9.69 C02 CXCL3 6.45 D02 IFNG 5.78 A06 CD40L 5.59 F02 MMP7 4.78 B06 CSF3 4.2 D05 IL1B 4.1 07 T MP1 3.79 D07 IL4 3.32 01 SERPIN 1 2.77 B12 CXCL1 2.65 F01 MMP2 2.4 indicates data missing or illegible when filed

TABLE-US-00010 TABLE 11 Position Gene Fold Change A07 CD − 65. 0 IT 6 −23. 0 A −144.31 C12 −122. B11 CT V −118.6 D12 IT A3 − 1.25 F06 PLAU −30.44 D03 IGF −27.01 F04 PD A −24. 8 D11 IT A2 −10.13 D04 IL −17. F11 RAC −16 B CT −15.86 04 T FA −11.3 10 VTN −10.43 B0 CTNN 1 −8.7 F09 PTEN −8.28 B04 COL A −8.16 2 TAT −8.12 A02 ACTC1 −7.78 04 ITGAV −7.62 B01 O 3 −6. A12 O −5.87 C0 F3 −5.42 A09 O −5.37 1 ITG −5. 03 TAGLN −5.02 F12 RHOA − .91 A03 ANGPT −4.56 A11 CO A1 − .43 A COL14A1 −4.25 06 T R −4.17 F08 PLG −4.06 07 IT −3.96 MAPK1 − . A10 COL1A2 −3.81 B03 COL5A2 −3.79 F0 MMP − .77 ITGA5 −3.76 COL5A1 −3.43 E10 MAPK3 −3.37 C10 O 2 −3.26 A01 ACTA2 −2.89 C05 EGFR −2.83 D10 IT A1 −2.8 D01 HGF −2.44 A05 CCL7 −2.3 G11 W P −2.21 B07 CT F −2.185 indicates data missing or illegible when filed

[0260] Next, wound healing PCR arrays revealed up- and down-regulated genes in 3D skin models in response to treatment with NB105-146 for thermal burn injury.

[0261] Total RNA from 3D skin models were characterised, and the relative expression levels for each gene in the two samples (burn (untreated) vs burned and treated with gel without mineral complex) are plotted against each other in the Scatter Plot. Results are shown in FIG. 8b and Tables 12 and 13.

[0262] Table 12 shows 12 genes that are up-regulated in response to treatment with NB105-146 relative to thermal burn injured (untreated) skin. Table 13 shows 57 genes that are down-regulated in response to NB105-146 treated versus untreated thermal burn injured skin.

TABLE-US-00011 TABLE 12 Position Gene Fold Change C09 FGF 26.19 A0 CD4 LG 10.89 D06 IL 6.65 C04 EGF 4.43 D03 GF 3. 3 06 C 3.35 B09 CT 3.16 08 B6 2. 5 C 8 F A 2.78 D02 IFNG 2.63 C 6 F13A1 2.61 B01 COL4A3 2.2 indicates data missing or illegible when filed

TABLE-US-00012 TABLE 13 Position Gene Fold Change − . −2 . C 2 −2 . −1 . −1 . CTNN −1 . 12 CXCL −1 .16 CXCL − . V − . C 2 − . − . MMP −64. T − .21 1 − . 2 − . 2 −43. 2 − . − . − . −39.4 −3 . − . COL − .4 A12 COL − .78 PL T −28.7 MAP −28. T −2 . −24. 1 STA −24.3 MAPK1 − 3.23 12 WN −2 . −2 .2 −2 .1 − . MM −1 . W −14. −14. −14. 8 C −1 . −1 . PTEN −1 .23 RAC −1 . C − . CO − . 8 −8. PL − . 4 −8.13 − .88 T − .52 CO − .72 D12 −4.66 −4. B CT −3. CoL −3. 2 −3. AN −3. indicates data missing or illegible when filed

[0263] Finally, wound healing PCR arrays revealed up- and down-regulated genes in 3D skin models in response to treatment with NB105-142 for thermal burn injuries. Total RNA from 3D skin models was characterised, and the relative expression levels for each gene in the two samples (WJ+Oriel vs Burn) were plotted against each other in the Scatter Plot. results are shown in FIG. 8c and Tables 14 and 15.

[0264] Table 14 shows 38 genes that are up-regulated in response to treatment with NB105-146 relative to thermal burn injured (untreated) skin. Table 15 shows 26 genes that are down-regulated in response to NB105-146 treated versus untreated thermal burn injured skin.

TABLE-US-00013 TABLE 14 Position Gene Fold Change D 5. B C . IL2 40. 7 C 38. C F A 33.31 8 29.08 A CDH 26. C 4 23. C 19. COL 1 . D 14.21 12.14 A02 ACTC1 1 . COL .4 8.99 A 7.86 CO . IL 7. C 5. 3 C12 H F 5.12 . CO 4. 7 A COL 4.71 TGF 4. 7 4.52 4. IL 4.44 COL 4. 4. 3.65 VTN . 12 IT .02 2. 7 11 C SV 2.7 2. 2. 2 1. T N 2.0 indicates data missing or illegible when filed

TABLE-US-00014 TABLE 15 Postion Gene Fold Change 95 C − 4 V −7722 CC − .27 −83. C −64. −60.18 C CXCL2 −21.0 12 CXCL1 −12.87 12 MMP1 −8.49 MMP −8.01 C CXCL5 −7. − . −6.02 C F7 −5.68 A C − . A ACT2 −4. 08 TM −3 − . − . MM −3 C −2.78 07 TIM −2.82 10 A −2. D 9 −2.22 CTNN −2.21 AUR −2.14 IT 1 −2.0 indicates data missing or illegible when filed

Example 4

[0265] Tissue dielectric constant of burned skin models was tested at time intervals following exposure to (treatment with) the mineral complex active ingredient (FIG. 7b) versus control (treatment with nothing) (FIG. 7a).

[0266] The invention has been described hereinbefore with reference to various embodiments. The description of these concrete embodiments only serves for explanation and a deeper understanding of the invention and is not to be considered as limiting the scope of the invention. Rather, the invention is defined by the annexed claims and the equivalents that are apparent to the one skilled in the art and which are in accordance with the general inventive concept.