Composite materials and methods of using the same for wound healing and antimicrobial applications

Abstract

The present teachings describe a composite material that is useful for wound healing and antimicrobial applications.

Description

[0001] COMPOSITE MATERIALS AND METHODS OF USING THE SAME FOR WOUND HEALING AND ANTIMICROBIAL APPLICATIONS

[0002] CROSS REFERENCE TO RELATED APPLICATIONS

[0003] This application claims the benefit of and priority to U.S. Patent Application No. 63 / 519,710 filed on August 15, 2023, the entire contents of which are incorporated by reference herein.

[0004] FIELD

[0005] The present teachings are related to wound healing and antimicrobial applications using a composite material containing titanium dioxide coated silver particles and titanium dioxide coated copper particles.

[0006] BACKGROUND

[0007] Medicine has evolved to advance wound prevention and treatment. Much of wound care is wound treatment, which involves promoting healing, preventing infections and / or treating already existing infections. Deciding on a treatment depends on the type of wound that a person has suffered, for example, an infection to severe bums. Although there are many types of wound dressings such as dry dressings, wet-to-dry dressings, chemical impregnated dressings, foam dressings, hydrogel dressings, and the like, expediting the healing process is always a concern and goal.

[0008] Accordingly, the art desires alternative materials for expediting wound healing as well as for use in antimicrobial applications.

[0009] SUMMARY

[0010] The present teachings provide a revolutionary technology of products (i.e. , composite materials) that can expedite wound healing of all types and that also can be useful for antimicrobial applications. That is, the composite materials of the present teachings provide bacteriostatic and / or bactericidal activity that facilitates the restoration of the natural balance of the localized area of treatment while preventing and treating infections.

[0011] More specifically, in one aspect, a composite material of the present teachings generally includes TiCh coated Ag (“(TiO2)n / Ag”) nanoparticles and TiCh coated Cu (“(TiO2)n / Cu”) nanoparticles where the (TiO2)n / Ag particles and the (TiO2)n / Cu nanoparticles are associated with a polymer such as polyethylene glycol in a vehicle. The (TiO2)n / Ag particles and the (TiO2)n / Cu nanoparticles can be further associated with another polymer such as chitosan. In various embodiments, the composite material also includes selenium. In some embodiments, the composite material includes zinc oxide.

[0012] In another aspect, the present teachings provide a method of making a composite material, where the method generally includes mixing TiO2, AgNCh and Cu(NO3)*3H2O in a first vehicle to form a first mixture; mixing a polymer in a second vehicle to form a second mixture; mixing the first mixture with the second mixture to form a final mixture; and exposing the final mixture to a reducing agent until less than about 15% of the Ag is Ag+thereby forming a composite material.

[0013] In various embodiments, the reducing agent comprises ultraviolet radiation, microwaves, or a combination thereof. In some embodiments, the methods include mixing Na2SeO4with the TiO2, AgNO3and Cu(NO3)*3H2O.

[0014] In another aspect, the present teachings provide a method of wound healing in a subject in need thereof comprising contacting an effective amount of a composite material as described herein with a wound of the subject to promote healing of the wound.

[0015] The present teachings also provide a method of treating or preventing an infection comprising contacting an effective amount of a composite material as described herein with an infection or an area of potential infection of a subject in need thereof to treat or prevent the infection.

[0016] The present teachings also include methods of treating surfaces of various materials of construction with the composite materials described herein to provide antimicrobial and disinfecting properties to the treated surfaces.

[0017] DESCRIPTION OF THE DRAWINGS

[0018] It should be understood that the drawings described below are for illustration purposes only. Like numerals generally refer to like parts. The drawings are not necessarily to scale, with emphasis generally being placed upon illustrating the principles of the present teachings. The drawings are not intended to limit the scope of the present teachings in any way.

[0019] FIG. 1 is a schematic diagram of a top view of a porcine model showing where the forty-eight (48) second degree bum wounds were created using five specially designed cylindrical brass rods. FIGS. 2-4 are pictures of an example of the process where wounds were sprayed with approximately 200 pL of test article from a spray bottle placed over the wounded area (FIGS. 2 and 3), which was then covered with a Tegaderm dressing (FIG. 4).

[0020] FIGS. 5 and 6 are pictures of an example of a second treatment group that had their wounds treated with 200 pL of silver sulfadiazine (SSD) each (FIG. 5), which were then covered with a Tegaderm dressing (FIG. 6).

[0021] FIG. 7 is a picture of an example of an untreated control wound that was without any treatment and covered only with a Tegaderm dressing.

[0022] FIG. 8 is a graph of Methicillin-resistant Staphylococcus aureus MRS A USA300 bacterial count after treatment application assessment at day 10, according to Example 6.

[0023] FIG. 9 is a graph of the percentage of epithelialization after day 4, day 7 and day 10, according to Example 6.

[0024] FIG. 10 is a graph of the epithelial thickness after day 4, day 7 and day 10, according to Example 6.

[0025] FIG. 11 is a graph of white cell infiltration after day 4, day 7 and day 10, according to Example 6.

[0026] FIG. 12 is a graph of granulation tissue formation after day 4, day 7 and day 10, according to Example 6.

[0027] FIG. 13 is a schematic diagram of an embodiment of a composite material of the present teachings including TiCEln / Ag nanoparticles, (TiChWCu nanoparticles, and (TiO2)n / Ag-Cu nanoparticles in the presence of Se and a polymer.

[0028] DETAILED DESCRIPTION

[0029] The present teachings describe composite materials that can expedite the healing of wounds such as burns as well as treating and / or preventing infection. The composite materials also can be an effective antimicrobial and useful in such applications, for example, disinfecting the surfaces of various materials.

[0030] The present teachings relate, in part, to compositions that can be used to improve wound healing such as burn recovery and to treat and / or reduce infection. The composite materials of the present teachings are effective for burn and wound disinfection and accelerated recovery. The composite materials are also useful to accelerate resolution of bedsores, and for the avoidance and accelerated eradication of acne, among many other uses.

[0031] The components of the composite materials of the present teachings include TiC associated with silver (Ag) nanoparticles such that numerous TiOz particles associate themselves with the Ag particles ((TiCbjn / Ag). (Here, although just meant for symbolism, n can be an integer greater than zero, for example, from about 5 to about 250.) Similarly, the components of the composite materials of the present teachings include (TiOz)n / Cu nanoparticles, which include numerous TiOz particles associated with the copper (Cu) particles. The components of the composite materials of the present teachings can also include numerous TiOz particles associated with the Ag-Cu aggregates or co-particles, which components can be referred to as (TiOzh / Ag-Cu nanoparticles. This arrangement in combination with selenium (Se) is depicted in FIG. 13.

[0032] Without wishing to be bound to any particular theory, it is believed that the composite materials can be envisioned as nano-semiconductor composites such that the composite materials described herein act via two different mechanisms. First, the composite materials can provide a nano silver, copper, and silver-copper “alloy” combined with selenium loaded with TiOz in a water based vehicle that can promote wound healing by the ability to generate reactive oxygen species (“ROS”) when exposed to natural light. ROS is an umbrella term for an array of derivatives of molecular oxygen that occur as a normal attribute of aerobic life. The composite material catalyzes disinfection on a surface when exposed to natural light, generating ROS. ROS play a critical role in wound healing by promoting angiogenesis (the formation of new blood vessels), stimulating cell proliferation and migration, and promoting collagen synthesis. That is, the ROS provides oxygen to the surfaces in which it contacts to promote a natural balance to the aerobic and anerobic species present for a healthy outcome.

[0033] Second, in addition to its ROS generating properties, the semiconductor interface can produce extracellular electron transfer (EET) in an aerobic dark environment, with no light, which shows antibacterial properties regardless of antibiotic resistance of the bacteria. EET is the process by which some microorganisms exchange intracellular electrons with an extracellular electron donor / acceptor, including naturally occurring metal compounds and artificial electrodes, across the cell membrane. Accordingly, an engineered semiconductor interface of the composite materials produces EET below the surface, in a dark environment, enhancing aerobic antibacterial action as a separate process not impacted by antibiotic resistance. As such, the composite materials work in natural light and on dark surfaces that make it perfect candidate for wound healing. The composite can help to inhibit the wide range of bacteria and fungi, which are cause of wound infections and often difficult to eradicate.

[0034] Another advantageous feature of the composite materials can be a cohesive biodegradable polymer such as polyethylene glycol that can assist with cell production by keeping the skin and wound hydrated with moisture on one hand, and can facilitate EET promoting signaling of the cells that indirectly promotes collagen production and heals the wound.

[0035] In sum, the composite materials described herein include photocatalytic materials that react not only at ultraviolet wavelengths but also on visible lights as the band gap energy of the nano photocatalyst composition is lowered by doping a noble metal of silver / copper electrodes in the middle and putting TiOz around the electrodes. In addition, the inclusion of selenium improves the electron exchange of the semiconductor material with cells and bacteria so the composite materials effect EET and ROS. In particular, the composition according to the present invention includes in the form of a TiOz-Ag-Cu electrode and a TiOz- Ag-Cu-Se nanocomposite having a doping structure in preparing a visible light-reactive TiOz photocatalyst composition on TiOz nanoparticles. The FIG. 13 shows the schematic view of the nano composite colloid in water-PEG solution.

[0036] Therefore, the composite materials of the present teachings can have wide photocatalytic activity from UV to visible light while have the lower bandgap and high potential for electron transmission with electron acceptors and can promote ROS production and also EET, both of which can inhibit bacteria or fungi infection and stimulate wound healing.

[0037] The composite materials described herein may further contain other noble metals such as gold (Au) and / or platinum (Pt), or a drug delivery polymer to enhance their functionality.

[0038] Definitions

[0039] To facilitate an understanding of the present invention, a number of terms and phrases are defined below.

[0040] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The abbreviations used herein have their conventional meaning within the chemical and biological arts. The chemical structures and formulae set forth herein are constructed according to the standard rules of chemical valency known in the chemical arts.

[0041] The terms “a’' and “an” as used herein mean “one or more” and include the plural unless the context is inappropriate.

[0042] In the application, where an element or component is said to be included in and / or selected from a list of recited elements or components, it should be understood that the element or component can be any one of the recited elements or components, or the element or component can be selected from a group consisting of two or more of the recited elements or components.

[0043] Further, it should be understood that elements and / or features of a composition or a method described herein can be combined in a variety of ways without departing from the spirit and scope of the present invention, whether explicit or implicit herein. For example, where reference is made to a particular compound, that compound can be used in various embodiments of compositions of the present invention and / or in methods of the present invention, unless otherwise understood from the context. In other words, within this application, embodiments have been described and depicted in a way that enables a clear and concise application to be written and drawn, but it is intended and will be appreciated that embodiments may be variously combined or separated without parting from the present teachings and invention(s). For example, it will be appreciated that all features described and depicted herein can be applicable to all aspects of the invention(s) described and depicted herein.

[0044] It should be understood that the expression “at least one of” includes individually each of the recited objects after the expression and the various combinations of two or more of the recited objects unless otherwise understood from the context and use. The expression “and / or” in connection with three or more recited objects should be understood to have the same meaning unless otherwise understood from the context.

[0045] The use of the term “include,” “includes,” “including,” “have,” “has,” “having,” “contain,” “contains,” or “containing,” including grammatical equivalents thereof, should be understood generally as open-ended and non-limiting, for example, not excluding additional unrecited elements or steps, unless otherwise specifically stated or understood from the context.

[0046] Where the use of the term “about” is before a quantitative value, the present invention also includes the specific quantitative value itself, unless specifically stated otherwise. As used herein, the term “about” refers to a ±10% variation from the nominal value unless otherwise indicated or inferred from the context.

[0047] At various places in the present specification, values are disclosed in groups or in ranges. It is specifically intended that the description include each and every individual subcombination of the members of such groups and ranges. For example, an integer in the range of 0 to 40 is specifically intended to individually disclose 0, 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, 35, 36, 37, 38, 39, and 40, and an integer in the range of 1 to 2O is specifically intended to individually disclose 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, and 20.

[0048] The use of any and all examples, or exemplary language herein, for example, “such as” or “including,” is intended merely to illustrate better the present invention and does not pose a limitation on the scope of the invention unless claimed. No language in the specification should be construed as indicating any non-claimed element as essential to the practice of the present invention.

[0049] The terms “individual,” “patient,” and “subject,” as used herein, are used interchangeably and include any animal, including mammals, preferably mice, rats, other rodents, rabbits, dogs, cats, swine, cattle, sheep, horses, or primates, and more preferably, humans. The compounds described in the disclosure can be administered to a mammal, such as a human, but can also be administered to other mammals such as an animal in need of veterinary treatment, for example, domestic animals (e. ., dogs, cats, and the like), farm animals (e.g., cows, sheep, pigs, horses, and the like) and laboratory animals (e.g., rats, mice, guinea pigs, and the like). The mammal treated in the methods described in the disclosure is preferably a mammal in which treatment, for example, of a skin cancer, is desired.

[0050] The phrases “pharmaceutically acceptable” and “pharmacologically acceptable,” as used herein, refer to compounds, molecular entities, compositions, materials, and / or dosage forms that do not produce an adverse, allergic or other untoward reaction when administered to an animal, or a human, as appropriate. For human administration, preparations should meet sterility, pyrogenicity, general safety and purity standards as required by FDA Office of Biologies standards.

[0051] As used herein, “pharmaceutically acceptable excipient” refers to a substance that aids the administration of an active agent to a subject and can be included in the compositions of the present invention without causing a significant adverse toxicological effect on the subject. Non-limiting examples of pharmaceutically acceptable excipients include water, NaCl, normal saline solutions, a phosphate buffered saline solution, emulsions (e.g., such as an oil / water or water / oil emulsions), lactated Ringer’s, normal sucrose, normal glucose, binders, fillers, disintegrants, lubricants, coatings, sweeteners, flavors, salt solutions (such as Ringer’s solution), alcohols, oils, gelatins, carbohydrates such as lactose, amylose or starch, fatty acid esters, hydroxypropylmethylcellulose, polyvinyl pyrrolidine, and colors, and the like. Such preparations can be sterilized and, if desired, mixed with auxiliary agents such as lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salts for influencing osmotic pressure, buffers, coloring, and / or aromatic substances and the like that do not deleteriously react with the compounds of the disclosure.

[0052] As used herein, the term “pharmaceutically acceptable carrier” refers to any of the standard pharmaceutical carriers, such as a phosphate buffered saline solution, water, emulsions e.g., such as an oil / water or water / oil emulsions), and various types of wetting agents. The compositions also can include stabilizers and preservatives. For examples of excipients, carriers, stabilizers and adjuvants, see Martin, Remington’s Pharmaceutical Sciences, 15th Ed., Mack Publ. Co., Easton, PA (1975).

[0053] The term “treating,” as used herein, includes any effect, for example, lessening, reducing, modulating, ameliorating, or eliminating, that results in the improvement of the condition, disease, disorder, and the like, including one or more symptoms thereof. Treating can be curing, improving, or at least partially ameliorating the disorder.

[0054] The phrase “therapeutically effective amount,” as used herein, refers to the amount of a compound (e.g., a disclosed compound) that will elicit the biological or medical response of a tissue, system, animal or human that is being sought by the researcher, veterinarian, medical doctor or other clinician. The compounds described in the disclosure can be administered in therapeutically effective amounts to treat a disease. A therapeutically effective amount of a compound can be the quantity required to achieve a desired therapeutic and / or prophylactic effect, such as an amount which results in lessening of a symptom of a condition such as a wound.

[0055] Throughout the description, where compositions and kits are described as having, including, or comprising specific components, or where processes and methods are described as having, including, or comprising specific steps, it is contemplated that, additionally, there are compositions and kits of the present invention that consist essentially of, or consist of, the recited components, and that there are processes and methods according to the present invention that consist essentially of, or consist of, the recited processing steps. As a general matter, compositions specifying a percentage are by weight unless otherwise specified.

[0056] Composite Materials

[0057] The effects described herein can be realized by contacting a composite material including TiOz coated silver ((TiOz)n / Ag) nanocomposites and TiOz coated copper ((TiO2)n / Cu) nanocomposites with a wound, a surface, or an area needing antibacterial activity. The ((TiChln / Ag) and (TiOz)n / Cu) nanocomposites are associated with a polymer in a vehicle, for example, water or an alcohol such as ethanol. In certain embodiments, the composite material includes selenium. In various embodiments, the composite material also includes TiOz coated Ag-Cu (TiOz / Ag-Cu) particles.

[0058] In some embodiments, the polymer of the composite material can be polyvinylpyrrolidone, polyvinyl alcohol, polyethylene glycol, and combinations thereof. Without wishing to be bound to any particular theory, it is believed that the polymer assists in disassociating the aggregates of the metal and metal oxide components in the composite materials. In addition, it is believed that a cohesive biodegradable polymer such as polyethylene glycol can promote cell production, keeping the skin and wound hydrated and moist while also assisting with EET signals to the cells, indirectly promoting collagen production and enhancing the healing process.

[0059] In various embodiments, the concentration of the TiOz in the composite material is about 50 mg / L to about 400 mg / L. In some embodiments, the concentration of the TiOz in the composite material is about 50 mg / L to about 350 mg / L, about 50 mg / L to about 300 mg / L, about 50 mg / L to about 250 mg / L, about 50 mg / L to about 200 mg / L, or about 50 mg / L to about 150 mg / L. In some embodiments, the concentration of the TiOz in the composite material is about 100 mg / L to about 400 mg / L, about 100 mg / L to about 350 mg / L, about 100 mg / L to about 300 mg / L, about 100 mg / L to about 250 mg / L, or about 100 mg / L to about 200 mg / L. In certain embodiments, the concentration of the TiOz in the composite material is about 150 mg / L to about 400 mg / L, about 150 mg / L to about 350 mg / L, about 150 mg / L to about 300 mg / L, about 150 mg / L to about 250 mg / L, or about 150 mg / L to about 200 mg / L. In particular embodiments, the concentration of the TiOz in the composite material is about 200 mg / L to about 400 mg / L, 200 mg / L to about 350 mg / L, about 200 mg / L to about 300 mg / L, or about 200 mg / L to about 250 mg / L. In various embodiments, the concentration of the TiOz in the composite material is about 200 mg / L to about 350 mg / L, or about 200 mg / L to about 325 mg / L, or about 250 mg / L to about 325 mg / L.

[0060] The TiCh generally is substantially anatase phase TiCh (e.g., at least about 80 %, at least about 85 %, at least about 90 %, or at least about 95 % , or at least about 99 % is this phase). The TiCb particles can have a mean diameter between about 0.5 nm and about 20 nm.

[0061] In various embodiments, the concentration of the Ag in the composite material is about 1 mg / L to about 60 mg / L. In some embodiments, the concentration of the Ag in the composite materials is about 5 mg / L to about 60 mg / L, about 5 mg / L to about 50 mg / L, about 5 mg / L to about 40 mg / L, about 5 mg / L to about 30 mg / L, about 5 mg / L to about 20 mg / L, or about 5 mg / L to about 15 mg / L. In certain embodiments, the concentration of the Ag in the composite material is about 10 mg / L to about 50 mg / L, about 10 mg / L to about 40 mg / L, about 10 mg / L to about 30 mg / L, about 10 mg / L to about 25 mg / L, or about 10 mg / L to about 20 mg / ml. In particular embodiments, the concentration of the Ag in the composite material is about 15 mg / L to about 50 mg / L, about 15 mg / L to about 40 mg / L, about 15 mg / L to about 30 mg / L, or about 15 mg / L to about 25 mg / L. In various embodiments, the concentration of the Ag in the composite material is about 10 mg / L to about 30 mg / L, or about 15 mg / L to about 30 mg / L.

[0062] The Ag in the composite material typically includes less than about 10 % of Ag+based on the total weight of silver in the composite material. In various embodiments, the Ag in the composite material includes less than about 20 % or less than about 15 % of Ag+based on the total weight of silver in the composite material. In some embodiments, the Ag in the composite material includes less than about 5 % Ag+based on the total weight of silver in the composite material. Ag particles can have a mean diameter of between about 15 nm and about 75 nm.

[0063] In various embodiments, the concentration of the Cu in the composite material is about 1 mg / L to about 10 mg / L. In some embodiments, the concentration of the Cu in the composite material is about 1 mg / L to about 7.5 mg / L, about 1 mg / L to about 5 mg / L, about 1 mg / L to about 4 mg / L, about 1 mg / L to about 3 mg / L, or about 1 mg / ml to about 2 mg / L. In certain embodiments, the concentration of the Cu in the composite material is about 2 mg / L to about 8 mg / L, about 2 mg / L to about 6 mg / L, or about 2 mg / L to about 4 mg / L. In particular embodiments, the concentration of the Cu in the composite material is about 1 mg / L to about 3 mg / L.

[0064] In various embodiments, the concentration of the Se in the composite material is about 0.01 mg / L to about 6 mg / L. In some embodiments, the concentration of the Se in the composite material is about 0.01 mg / L to about 4 mg / L, about 0.01 mg / L to about 3 mg / L, about 0.01 mg / L to about 2 mg / L, or about 0.01 mg / L to about 1 mg / L. In certain embodiments, the concentration of the Se in the composite material is about 0.1 mg / L to about 4 mg / L, about 0.1 mg / L to about 3 mg / L, about 0. 1 mg / L to about 2 mg / L, or about 0.1 mg / L to about 1 mg / L. In particular embodiments, the concentration of the Se in the composite material is about 1 mg / L to about 4 mg / L, about 1 mg / L to about 3 mg / L, or about 1 mg / L to about 2 mg / L. In various embodiments, the concentration of the Se in the composite material is about 1.5 mg / L.

[0065] The concentrations of the metals in the composite materials can be at different ratios in various embodiments. The ratio of Cu to Se can be about 1 : 1 to about 3 : 1 , for example, about 2: 1. The ratio of Ag to Se can be about 10:1 to about 20: 1 , for example, 11: 1 to about 18: 1, including 11:1 and 18:1. The ratio of TiCL to Se can be about 125: 1 to about 250:1, for example, 167 : 1 to about 211:1, including 167 : 1 and 211 :1. Where selenium is not present in the composite material, the ratio of Ag to Cu can be about 5:1 to about 15: 1, for example, about 8: 1 to about 10:1. The ratio of TiCL to Cu can be about 100: 1 to about 160:1, for example, about 120:1 to about 150:1.

[0066] In various embodiments, the concentration of the polymer in the composite material is about 300 mg / L to about 5000 mg / L. In some embodiments, the concentration of the polymer in the composite material is about 300 mg / L to about 4000 mg / L, about 300 mg / L to about 3000 mg / L, about 300 mg / L to about 2500 mg / L, about 300 mg / L to about 2000 mg / L, about 300 mg / L to about 1500 mg / L, about 300 mg / L to about 1000 mg / L or about 300 mg / L to about 750 mg / L. In particular embodiments, the concentration of the polymer in the composite material is about 350 mg / L to about 1500 mg / L, about 350 mg / L to about 750 mg / L, or about 750 mg / L to about 1500 mg / L. It should be understood that when referring to the polymer, the polymer encompasses one or more polymers that may be present in the composite material.

[0067] In various embodiments, the composite material includes about 93 % to about 99.9 % by weight of the vehicle based on the total weight of the composite material. In certain embodiments the vehicle comprises water. In some embodiments, the vehicle is water. In various embodiments, the vehicle includes one or more of water, ethylene glycol, methylene glycol, diethylene glycol, an acid, and combinations thereof.

[0068] In some embodiments, the composite material further includes an acid. The acid can be acetic acid, hydrochloric acid, or combinations thereof. In some embodiments, the concentration of acetic acid in the composite material is about 0.001% to about 1 % by weight based on the total weight of the composite material. In some embodiments, the concentration of acetic acid in the composite material is about 0.01% to about 0.5%, about 0.01% to about 0.25%, or from about 0.01% to about 0.1% based on the total weight of the composite material. In particular embodiments, the concentration of hydrochloric acid in the composite material is about 0.0001% to about 0.001%, about .0001% to about 0.0005%, or about 0.00025% about 0.0005% based on the total weight of the composite material.

[0069] In certain embodiments, the composite material further includes zinc oxide (ZnO). In some embodiments, the concentration of the ZnO in the composite material is about 0.001 mg / L to about 75 mg / L. In various embodiments, the concentration of the ZnO in the composite material is about 0.01 mg / L to about 50 mg / L about 0. 1 mg / L to about 50 mg / L, about 0.1 mg / L to about 40 mg / L about 0.1 mg / L to about 30 mg / L, about 0.1 mg / L to about 20 mg / L, about 0.1 mg / L to about 10 mg / L, or about 0.1 mg / L to about 5 mg / L. In some embodiments, the concentration of ZnO in the composite material is about 1 mg / L to about 50 mg / L, about 1 mg / L to about 40 mg / L, about 1 mg / L to about 30 mg / L, about 1 mg / L to about 20 mg / L, about 1 mg / L to about 10 mg / L, or from about 1 mg / L to about 5 mg / L. In particular embodiments, the concentration of ZnO in the composite material is about 5 mg / L to about 50 mg / L, about 5 mg / mL to about 30 mg / L, about 5 mg / L to about 20 mg / L, or from about 5 mg / L to about 10 mg / L.

[0070] In particular embodiments, the composite material includes chitosan. The weight ratio of the chitosan to the metals and metal oxides of the composite material can be in a range of about 0:1 to about 10:1. The weight ratio of the chitosan to the polymer can be in a range of about 0:1 to about 7:1. The concentration of the chitosan can be about 1 mg / L to about 2500 mg / L.

[0071] In particular embodiments, a composite material includes titanium dioxide coated silver particles; titanium dioxide coated copper particles; polyvinylpyrrolidone, and water. In certain embodiments, a composite material includes titanium dioxide coated silver particles; titanium dioxide coated copper particles; selenium; polyvinylpyrrolidone, and water. In such embodiments, the composite material can also include one or more of zinc oxide, acetic acid, and chitosan. The composite material also can include polyethylene glycol.

[0072] Depending on the application, the composite material can further contain precious or noble metals such as gold (Au) and / or platinum (Pt) or transition metal components such as nickel (Ni) and cobalt (Co) in addition to Ag and Cu.

[0073] The pH of the composite material typically is in the range of about 3 to about 6. In some embodiments the pH of the composite material is in the range of about 4 to about 5. The pH of the composite material can be adjusted using the appropriate acid or base.

[0074] Forms of Composite Materials

[0075] A composite material can be prepared in any of a variety of forms for use such as for administering an active agent to a patient, who may be in need thereof. For example, the composite materials of the present teachings can be formulated for administration in liquid or cream-like form, including those adapted for the following: (1) topical administration, for example, as a cream, ointment, or a controlled-release patch or spray applied to the skin; and (2) intravaginal or intrarectal administration, for example, as a pessary, cream or foam.

[0076] In various embodiments, the composite material is in the form of a spray. In certain embodiments, the composite material is positioned or mounted on a solid support, for example, as a solution or a cream, or other type of delivery types discussed herein.

[0077] The composite materials of the present teachings can be topical formulations. The topical formulations may be formulated as a liquid, a solution, an emulsion, a cream, a lotion, a suspension, a triturate, a gel, a jelly, a foam, a paste, an ointment, a shampoo, an adhesive, a patch, a foam, an aerosol foam, or the like. In certain embodiments, the topical formulations of the present teachings may be formulated as a liquid. In certain embodiments, the topical formulations of the present teachings may be formulated as a solution. In certain embodiments, the topical formulations of the present teachings may be formulated as an emulsion. In certain embodiments, the topical formulations of the present teachings may be formulated as a cream. In certain embodiments, the topical formulations of the present teachings may be formulated as a lotion. In certain embodiments, the topical formulations of the present teachings may be formulated as a suspension. In certain embodiments, the topical formulations of the present teachings may be formulated as a triturate. In certain embodiments, the topical formulations of the present teachings may be formulated as a gel. In certain embodiments, the topical formulations of the present teachings may be formulated as a jelly. In certain embodiments, the topical formulations of the present teachings may be formulated as a foam. In certain embodiments, the topical formulations of the present teachings may be formulated as a paste. In certain embodiments, the topical formulations of the present teachings may be formulated as an ointment. In certain embodiments, the topical formulations of the present teachings may be formulated as a shampoo. In certain embodiments, the topical formulations of the present teachings may be formulated as an adhesive. In certain embodiments, the topical formulations of the present teachings may be formulated as a patch. In certain embodiments, the topical formulations of the present teachings upon application to a skin of a subject may form a patch.

[0078] Ointments, as is well known in the art of pharmaceutical formulation, are semisolid preparations that are typically based on petrolatum or other petroleum derivatives. The specific ointment base to be used, as will be appreciated by those skilled in the art, is one that will provide for optimum drug delivery, and, preferably, will provide for other desired characteristics as well, e.g., emolliency or the like. As with other carriers or vehicles, an ointment base should be inert, stable, nonirritating and nonsensitizing. As explained in Remington: The Science and Practice of Pharmacy, 19th Ed. (Easton, Pa.: Mack Publishing Co., 1995), at pages 1399-1404, ointment bases may be grouped in four classes: oleaginous bases; emulsifiable bases; emulsion bases; and water-soluble bases. Oleaginous ointment bases include, for example, vegetable oils, fats obtained from animals, and semisolid hydrocarbons obtained from petroleum. Emulsifiable ointment bases, also known as absorbent ointment bases, contain little or no water and include, for example, hydroxystearin sulfate, anhydrous lanolin, and hydrophilic petrolatum. Emulsion ointment bases are either water-in-oil (W / O) emulsions or oil-in-water (O / W) emulsions, and include, for example, cetyl alcohol, glyceryl monostearate, lanolin, and stearic acid. In embodiments, water-soluble ointment bases are prepared from polyethylene glycols of varying molecular weight; again, see Remington: The Science and Practice of Pharmacy for further information.

[0079] Creams, as also well known in the art, are viscous liquids or semisolid emulsions, either oil-in-water or water-in-oil. Cream bases are water- washable, and contain an oil phase, an emulsifier, and an aqueous phase. The oil phase, also called the “internal” phase, is generally included of petrolatum and a fatty alcohol such as cetyl or stearyl alcohol. The aqueous phase usually, although not necessarily, exceeds the oil phase in volume, and generally contains a humectant. The emulsifier in a cream formulation is generally a nonionic, anionic, cationic, or amphoteric surfactant.

[0080] Gels are semisolid, suspension-type systems. Single-phase gels contain organic macromolecules distributed substantially uniformly throughout the carrier liquid, which is typically aqueous, but also contains an alcohol and, optionally, an oil. In embodiments, “organic macromolecules,” i.e., gelling agents, are crosslinked acrylic acid polymers such as the “carbomer” family of polymers, e.g., carboxypolyalkylenes that may be obtained commercially under the CARBOPOL™ trademark. Hydrophilic polymers such as polyethylene oxides, polyoxyethylene-polyoxypropylene copolymers, and polyvinyl alcohol; cellulosic polymers such as hydroxypropyl cellulose, hydroxyethyl cellulose, hydroxypropyl methylcellulose, hydroxypropyl methylcellulose phthalate, and methyl cellulose; gums such as tragacanth and xanthan gum; sodium alginate; and gelatin may also be included. In order to prepare a uniform gel, dispersing agents such as alcohol or glycerin can be added, or the gelling agent can be dispersed by trituration, mechanical mixing, or stirring, or combinations thereof.

[0081] Lotions are preparations to be applied to the skin surface without friction, and are typically liquid or semiliquid preparations in which solid particles, including the active agent, are present in a water or alcohol base. Lotions are usually suspensions of solids, and preferably, for the present purpose, include a liquid oily emulsion of the oil-in-water type. In embodiments, lotions are used for treating large body areas, because of the ease of applying a more fluid composition. It is generally necessary that the insoluble matter in a lotion be finely divided. Lotions will typically contain suspending agents to produce better dispersions as well as compounds useful for localizing and holding the active agent in contact with the skin, e.g., methylcellulose, sodium carboxymethylcellulose, or the like.

[0082] Pastes are semisolid dosage forms in which the compound of Formula (I), or a pharmaceutically acceptable salt thereof, is suspended in a suitable base. Depending on the nature of the base, pastes are divided between fatty pastes or those made from a single-phase aqueous gels. The base in a fatty paste is generally petrolatum, hydrophilic petrolatum, or the like. The pastes made from single-phase aqueous gels generally incorporate carboxymethylcellulose or the like as a base.

[0083] Formulations may also be prepared with liposomes, micelles, and microspheres. Liposomes are microscopic vesicles having a lipid wall including a lipid bilayer, and can be used as drug delivery systems herein as well. Generally, liposome formulations are poorly soluble or insoluble pharmaceutical agents. Liposomal preparations for use in the instant disclosure may include cationic (positively charged), anionic (negatively charged), and neutral preparations. Cationic liposomes are readily available. For example, N[l-2,3- dioleyloxy)propyl]-N,N,N-triethylammonium (DOTMA) liposomes are available under the trade name LIPOFECTIN™. (ThermoFisher). Similarly, anionic and neutral liposomes are readily available as well, e.g., from Avanti Polar Lipids (Birmingham, Ala.), or can be easily prepared using readily available materials. Such materials include phosphatidyl choline, cholesterol, phosphatidyl ethanolamine, dioleoylphosphatidyl choline (DOPC), dioleoylphosphatidyl glycerol (DOPG), and dioleoylphoshatidyl ethanolamine (DOPE), among others. These materials can also be mixed with DOTMA in appropriate ratios. Methods for making liposomes using these materials are well known in the art.

[0084] Micelles are known in the art as included of surfactant molecules arranged so that their polar head groups form an outer spherical shell, while their hydrophobic, hydrocarbon chains are oriented towards the center of the sphere, forming a core. Micelles form in an aqueous solution containing surfactant at a high enough concentration so that micelles naturally result. Surfactants useful for forming micelles include, but are not limited to, potassium laurate, sodium octane sulfonate, sodium decane sulfonate, sodium dodecane sulfonate, sodium lauryl sulfate, docusate sodium, decyltrimethylammonium bromide, dodecyltrimethylammonium bromide, tetradecyltrimethylammonium bromide, tetradecyltrimethylammonium chloride, dodecylammonium chloride, polyoxyl 8 dodecyl ether, polyoxyl 12 dodecyl ether, nonoxynol 10, and nonoxynol 30. Micelle formulations can be used in conjunction with the present teachings either by incorporation into the reservoir of a topical or transdermal delivery system, or into a formulation to be applied to the body surface.

[0085] Microspheres, similarly, may be incorporated into the present formulations and drug delivery systems. Like liposomes and micelles, microspheres essentially encapsulate a drug or drug-containing formulation. Microspheres are generally, although not necessarily, formed from synthetic or naturally occurring biocompatible polymers, but may also be included of charged lipids such as phospholipids. Preparation of microspheres is well known in the art and described in the pertinent texts and literature.

[0086] Various additives, known to those skilled in the art, may be included in the topical formulations. For example, solvents, including relatively small amounts of alcohol, may be used to solubilize certain formulation components. In embodiment, the formulation includes a suitable enhancer, e.g., but are not limited to, ethers such as diethylene glycol monoethyl ether (available commercially as TRANSCUTOL™) and diethylene glycol monomethyl ether; surfactants such as sodium laurate, sodium lauryl sulfate, cetyltrimethylammonium bromide, benzalkonium chloride, Poloxamer (231, 182, 184), Tween (20, 40, 60, 80), and lecithin (U.S. Patent No. 4,783,450); alcohols such as ethanol, propanol, octanol, benzyl alcohol, and the like; polyethylene glycol and esters thereof such as polyethylene glycol monolaurate (PEGML); amides and other nitrogenous compounds such as urea, dimethylacetamide (DMA), dimethylformamide (DMF), 2-pyrrolidone, l-methyl-2- pyrrolidone, ethanolamine, diethanolamine, and triethanolamine; terpenes; alkanones; and organic acids, particularly citric acid and succinic acid. AZONETM and sulfoxides such as DMSO and CIO MSO may also be used.

[0087] The present formulations may also include conventional additives such as opacifiers, antioxidants, fragrance, colorant, gelling agents, thickening agents, stabilizers, surfactants, and the like. Other agents may also be added, such as antimicrobial agents, to prevent spoilage upon storage, i.e., to inhibit growth of microbes such as yeasts and molds. Suitable antimicrobial agents are typically selected from the group consisting of the methyl and propyl esters of p-hydroxybenzoic acid (i.e., methyl and propyl paraben), sodium benzoate, sorbic acid, imidurea, and combinations thereof.

[0088] The formulations may also contain irritation-mitigating additives to minimize or eliminate the possibility of skin irritation or skin damage resulting from the pharmacologically active base or other components of the composition. Suitable irritationmitigating additives include, for example: a-tocopherol; monoamine oxidase inhibitors, particularly phenyl alcohols such as 2 -phenyl- 1 -ethanol; glycerin; salicylic acids and salicylates; ascorbic acids and ascorbates; ionophores such as monensin; amphiphilic amines; ammonium chloride; N-acetylcysteine; cis-urocanic acid; capsaicin; and chloroquine. The irritant-mitigating additive, if present, may be incorporated into the present formulations at a concentration effective to mitigate irritation or skin damage, typically representing not more than about 20 wt. %, more typically not more than about 5 wt. %, of the composition.

[0089] Amounts of a disclosed compound as described herein in a formulation may vary according to factors such as the disease state, age, sex, and weight of the individual. Dosage regimens may be adjusted to provide the optimum therapeutic response. For example, a single bolus may be administered, several divided doses may be administered over time or the dose may be proportionally reduced or increased as indicated by the exigencies of the therapeutic situation. It is especially advantageous to formulate parenteral compositions in dosage unit form for ease of administration and uniformity of dosage. Dosage unit form as used herein refers to physically discrete units suited as unitary dosages for the mammalian subjects to be treated; each unit containing a predetermined quantity of active compound calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier.

[0090] The specification for the dosage unit forms are dictated by and directly dependent on (a) the unique characteristics of the compound selected and the particular therapeutic effect to be achieved, and (b) the limitations inherent in the art of compounding such an active compound for the treatment of sensitivity in individuals.

[0091] Therapeutic compositions typically must be sterile and stable under the conditions of manufacture and storage. The composition can be formulated as a solution, microemulsion, liposome, or other ordered structure suitable to high drug concentration. The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol, and the like), and suitable mixtures thereof. The proper fluidity can be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants. In many cases, it will be preferable to include isotonic agents, for example, sugars, polyalcohols such as mannitol, sorbitol, or sodium chloride in the composition.

[0092] The compounds can be administered in a time release formulation, for example in a composition which includes a slow release polymer. The compounds can be prepared with carriers that will protect the compound against rapid release, such as a controlled release formulation, including implants and microencapsulated delivery systems. Biodegradable, biocompatible polymers can be used, such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, polylactic acid and polylactic, polyglycolic copolymers (PLG). Many methods for the preparation of such formulations are generally known to those skilled in the art.

[0093] In some embodiments, a compound can be formulated with one or more additional compounds that enhance the solubility of the compound. In certain embodiments, composite materials described herein can be administered in combination with one or more additional therapeutic agents to treat a disorder described herein.

[0094] Methods of Making the Composite Materials

[0095] In another aspect, the present teachings generally provide a method of making a composite material. The methods can generally comprise mixing TiCT Ag salts (e.g., AgNCh), and Cu salts (e.g., Cu(NOs)*3H2O) in a first vehicle to form a first mixture that is mixed with a second mixture containing a polymer in a second vehicle to form a final mixture; and exposing the final mixture to a reducing agent until less than about 20% of the Ag is Ag+thereby forming a composite material.

[0096] The composition for a composite material of the present teachings can comprise an first mixture containing TiOz, Ag, Cu in a first vehicle; and a second mixture containing a polymer in a second vehicle. The first vehicle with TiC can include an acid such as hydrochloric acid to initially solubilize the Ag. Other components as described herein can be present in the appropriate mixture prior to forming a composite material of the present teachings. For example, the first mixture can also contain selenium salts (e.g., Na2SeO4), or other precious metal salts as described herein.

[0097] The two mixtures are mixed together and then exposed to a reducing agent such as microwaves, ultraviolet (UV) radiation (e.g., UV-C), or both until the amount of Ag present is less than about 15% Ag+, or less than about 10% Ag+, or less than about 5% Ag+based on the total amount of silver present. The amount of Ag+ present can be determined by using UV spectrometry, for example, at a wavelength of about 415 nm to about 430 nm.

[0098] The total amount of the first vehicle and the second vehicle (also referred to herein as the “vehicle” of the composite material) in the composite material can be between about 95% to about 99.9 % by weight based on the total weight of the composite material. In various embodiments, the composite material includes about 95% to about 99.9% by weight of the first vehicle and the second vehicle based on the total weight of the composite material. In some embodiments, the composite material includes about 80 to about 99.9% by weight of the first vehicle and the second vehicle based on the total weight of the composite material.

[0099] In various embodiments, the reducing agent comprises ultraviolet radiation (e.g., UV- C), micro waves, or a combination thereof. In certain embodiments the mixture is exposed to a reducing agent until less than about 15% or less than about 10% by weight of the Ag is Ag+. Methods of Using the Composite Materials

[0100] In another aspect, the present teachings provide a method of wound healing in a subject in need thereof comprising contacting the composite material as described herein with a wound of the subject to promote healing of the wound. The present teachings also provide a method of treating or preventing an infection comprising contacting the composite material as described herein with an infection or an area of potential infection of the subject to treat or prevent the infection. The various delivery types and vehicles as discussed herein elsewhere and will not be expanded upon other than to incorporate those teachings into the methods of using the composite materials.

[0101] The methods can be effective for treating a variety of conditions and wound healing where the methods can treat a condition selected from the group consisting of first, second, or third degree burns; a compromise of the superficial layer, keratin layer, epithelial layer or deeper layers of skin; a disease that cause compromise of the superficial or deep layers of skin; skin colonization with any bacterial, microbial or fungal species; a compromise of superficial layers, internal epithelium, including superficial or deep layers of the gastrointestinal system; a compromise of internal epithelium or superficial and deeper layers of the anorectal system; a compromise of superficial and internal layers of the oral and dental mucosa; surgical debridement or intervention; pre- or post-surgical wound in any part of the body; an intervention or prevention of facial, scalp, oral, periportal, anal, peri-anal or any mucosal infection; a puncture; a rash; and an acneiform disorder, including for treating first, second, or third degree burns, a compromise of the superficial layer, keratin layer, epithelial layer or deeper layers of skin, a disease that can compromise the superficial or deep layers of skin, skin colonization with any bacterial, microbial or fungal species, a compromise of superficial layers, internal epithelium including superficial or deep layers of the gastrointestinal system, a compromise of internal epithelium or superficial and deeper layers of the anorectal system, a compromise of superficial and internet layers of the oral and dental mucosa, surgical debridement or intervention or pre- or post-surgical wound in any part of the body, an intervention or prevention of facial, scalp, oral, periporal, anal, peri-anal or any mucosal infection, wound, puncture, bum, rash, or acneiform disorder.

[0102] The composite materials described herein also can be used to coat a medical device to be used on or in the body of a subject such as a human. Such devices can include scalpels and clips to pacemakers to other implantable devices. The present teachings also provides a combination therapy, which includes the administration of a composite material described herein and a second agent as part of a specific treatment regimen intended to provide the beneficial effect from the co-action of these therapeutic agents. The beneficial effect of the combination may include pharmacokinetic or pharmacodynamic co-action resulting from the combination of therapeutic agents.

[0103] Examples

[0104] Example 1. Production of Composite Material

[0105] Example 1A. One liter (L) of composite material was prepared by preparing a first mixture by mixing polyvinylpyrrolidone (PVP) (380 mg) and polyethylene glycol (PEG) (380 mg) with about 987.5 mb distilled water at ambient temperature.

[0106] A second mixture is prepared by mixing TiO2 solution (TiO (250 mg) dispersed in 18.5% HC1 (0.4625 mL HC1 in 2.0375 mb distilled water), AgNCh (25 mg / L dissolved in 4.975 mL distilled water), cupric nitrate trihydrate (Cu(NO3)2*3H2O) (10.8 mg dissolved in 4.99 mL distilled water) and sodium selena te (Na2SeO4) (3.16 mg dissolved in 4.997 mL distilled water) for about 24 hours at ambient temperature. The first mixture is mixed with the second mixture to provide a final mixture of about 1 L. The final mixture was mixed and exposed to UV radiation until the conversion of Ag+ to Ag became about 90% (i.e., less than about 10 % by weight of Ag+remains in the final mixture).

[0107] The amount of effective material considered for this formulation’s concentration is about 270 mg / mL (i.e., 250 mg / L TiCL + 15.87 mg / mL Ag + 2.84 mg / mL Cu + 1.44 mg / mL Se).

[0108] Example IB. One liter (L) of composite material was prepared by preparing a first mixture by mixing polyvinylpyrrolidone (PVP) (750 mg) and polyethylene glycol (PEG) (750 mg) with about 981.8 mL distilled water at ambient temperature.

[0109] A second mixture is prepared by mixing TiCL solution (TiO (316 mg) dispersed in 18.5% HC1 (0.592 mL HC1 in 2.608 mL distilled water), AgNOa (42.48 mg / L dissolved in 4.958 mL distilled water), cupric nitrate trihydrate (Cu / NOsh^HsO) (10.8 mg dissolved in 4.99 mL distilled water) and sodium selenate (Na2SeO4) (3.16 mg dissolved in 4.997 mL distilled water) for about 24 hours at ambient temperature. The first mixture is mixed with the second mixture to provide a final mixture of about 1 L. The final mixture was mixed and exposed to UV radiation until the conversion of Ag+ to Ag became about 90% (i.e., less than about 10 % by weight of Ag+remains in the final mixture).

[0110] The amount of effective material considered for this formulation’s concentration is about 347 mg / mL (i.e., 316 mg / L TiCb + 26.97 mg / mL Ag + 2.84 mg / mL Cu + 1 .44 mg / mL Se).

[0111] Example 1C. One liter (L) of composite material was prepared by preparing a first mixture by mixing polyvinylpyrrolidone (PVP) (380 mg) with about 988 mL distilled water at ambient temperature.

[0112] A second mixture is prepared by mixing TiCL solution (TiOz (200 mg) dispersed in 14.8% HC1 (0.37 mL HC1 in 2.13 mL distilled water), AgNCh (20.0 mg / L dissolved in 4.98 mL distilled water), and cupric nitrate trihydrate (Cu(NO3)ze3H2O) (5.4 mg dissolved in 4.995 mL distilled water) for about 24 hours at ambient temperature. The first mixture is mixed with the second mixture to provide a final mixture of about 1 L. The final mixture was mixed and exposed to UV radiation until the conversion of Ag+ to Ag became about 90% (i.e., less than about 10 % by weight of Ag+remains in the final mixture).

[0113] The amount of effective material considered for this formulation’s concentration is about 214 mg / mL (i.e., 200 mg / L TiCL + 12.7 mg / mL Ag + 1.42 mg / mL Cu).

[0114] Example 2. In-vitro Microbiological Susceptibility Tests

[0115] Example 2a. Cutibacterium acnes

[0116] Composite material concentration received: 347.25 mg / mL (concentration of effective materials in formulation; prepared according to Example IB)

[0117] Composite material concentrations tested: 100%, 50%, 25%, 12.5%, 6.25%, 3.13%, 1.56%, 0.78%, 0.39%, 0.2% & 0.1% of formulation

[0118] Minimum Inhibitory Concentration (MIC) assays by microbroth dilution method determine the lowest concentration of an antimicrobial agent that prevents visible growth of a microorganism in broth after overnight incubation. This precise method of susceptibility testing allows for quantification of the exact concentration of the antimicrobial agent (or client selected test article) needed to inhibit growth of microorganisms. For C. acnes, the media used for MIC assay is Brucella broth.

[0119] Minimum Bactericidal Concentration (MBC) assays determine the lowest concentration of an antimicrobial agent required to achieve f bacterial killing, defined as a 99.9% reduction in the initial inoculum. To establish MBC values, the total number of viable cell counts in each well was determined by plating 10 pL of culture from the broth wells for all wells above the MIC and monitored for bacterial growth.

[0120] The MIC test shows the lowest level of antimicrobial agent required to inhibit the growth of microorganisms, while MBC shows the lowest level of antimicrobial agent that causes the death of microorganisms. This means that even if a specific MIC exhibits a good inhibitory effect, inoculating microorganism on agar may still cause proliferation, because the antimicrobial agent at this concentration will not cause the complete death of the organism.

[0121] MBC:MIC ratios are usually interpreted using established criteria such that the antimicrobial agent is considered bactericidal if the MBC:MIC ratio is < 4, and bacteriostatic when the ratio is > 4.

[0122] Results: MIC by broth microdilution assay (visual): 25% (equivalent to 86.81 mg / L)

[0123] MIC by broth microdilution assay (spectrophotometry): Not determined due to the interference with Brucella broth media color.

[0124] MBC: 86.61 mg / mL (No growth observed in subculture done after exposure to 25%, 50% and 100% formulation of Example IB at 48 hrs)

[0125] MBC: MIC = 4

[0126] Example 2b. Pseudomonas sp .

[0127] Composite material concentration received: 347.25 mg / mL (concentration of effective materials in formulation; prepared according to Example IB)

[0128] Composite material concentrations tested: 100%, 50%, 25%, 12.5%, 6.25%, 3.13%, 1.56%, 0.78%, 0.39%, 0.2% & 0.1% of formulation

[0129] Results: MIC by broth microdilution assay (visual): 25% (86.61 mg / mL)

[0130] MIC by broth microdilution assay (spectrophotometry): 25% (86.61 mg / mL) MBC: 195 ppm (No growth observed in subculture done after exposure to 345 mg / mL composite material at 24 hrs and 72 hrs)

[0131] MBC: MIC = 3.9

[0132] Example 2c. Methicillin-resistant Staphylococcus aureus (MRSA)

[0133] Composite material concentration received: 347.25 mg / mL (concentration of effective materials in formulation; prepared according to Example IB)

[0134] Composite material concentrations tested: 100%, 50%, 25%, 12.5%, 6.25%, 3.13%, 1.56%, 0.78%, 0.39%, 0.2% & 0.1% of formulation

[0135] Results: MIC by broth microdilution assay (visual): 25% (86.61 mg / mL)

[0136] MIC by broth microdilution assay (spectrophotometry): 50% (173.63 mg / mL)

[0137] MBC: Growth observed in the subculture done after exposure to composite material. Bactericidal activity not observed against MRSA.

[0138] MBC: MIC = N / A

[0139] Example 2d. Candida auris

[0140] Composite material concentration received: 347.25 mg / mL (concentration of effective materials in formulation; prepared according to Example IB)

[0141] Composite material concentrations tested: 85%, 50%, 25%, 12.5%, 6.25%, 3.13%, 1.56%, 0.78%, 0.39%, 0.2% & 0.1% of formulation

[0142] Results: MIC by broth microdilution assay (visual): 50% (173.63 mg / mL)

[0143] MIC by broth microdilution assay (spectrophotometry): Not determined

[0144] MFC: Although no growth was observed at 24 hrs in the subculture done after exposure to 85% of formulation of composite material (295.16 mg / mL), growth was observed at 48 hrs

[0145] MFC: MIC = N / A Example 3. Determination of the Effects of Composite Material on Second Degree Burn Wounds Using a Porcine Model

[0146] OBJECTIVE

[0147] The objective of this study was to determine the healing and microbial effects of a new formulation using a porcine second degree burn wound model. The composite material formulation used was that prepared in Example 1 A and also referred to herein as the “spray formulation.”

[0148] MATERIALS AND METHODS

[0149] Experimental Animals

[0150] A porcine model was used for our experimental research due to the morphological similarities between swine skin and human skin. One female animal specific pathogen free (B. G. Looper Farm 4673 Petra Mill Road Granite Falls, NC 28630) pig weighing 40-45 kg was kept in house for at least 5 days prior to initiating the experiment to allow the animals to acclimatize. The animal was fed a basal diet ad libitum and housed individually in an animal facilities (meeting American Association for Accreditation of Laboratory Animal Care [AAALAC] accredited) with controlled temperature (19-21 °C) and lighting (12 h / 12 h LD).

[0151] Wounding Technique

[0152] Forty-eight (48) second degree bum wounds were created using five specially designed cylindrical brass rods weighing 358 g each that were heated in a boiling water bath to 100 °C (see FIG. 1). A rod was removed from the water bath and wiped dry before it was applied to the skin surface to prevent water droplets from creating a steam burn on the skin. The brass rod was held at a vertical position on the skin (6 seconds), with all pressure supplied by gravity, in order to make a bum wound that is 8.5 mm in diameter and approximately 0.8 mm deep. Immediately after burning, the roof of the bum blister was removed with a sterile spatula. The wounds were separated from one another by about 5-7 cm of un wounded skin. After wounding, the wounded area was divided in two sections: one for wound healing analysis (thirty-six uninoculated wounds were assigned to 3 groups with 12 wounds each) and the other for microbiology analysis (12 wounds were assigned to 3 groups with 4 wounds each), respectively as shown in FIG. 1. All wounds were treated within 20 minutes after wounding and / or wounding and infection, covered with a polyurethane film dressing (Tegaderm™; 3M, St. Paul, MN), secured with surgical tape and the entire animal loosely wrapped with self-adherent bandages (Coban™; 3M, St. Paul, MN). Wound Inoculation

[0153] A pathogenic strain of Methicillin-resistant Staphylococcus aureus MRSA USA300 (MRS A USA300) was utilized for the inoculum preparation. The challenge inoculum suspension was prepared by scraping the overnight grown bacteria from a culture plate into 4.5 mL of normal saline. This resulted in a suspension concentration of approximately IO10colony forming units / mL (CFU / mL). Serial dilutions were made until a concentration of 106CFU / mL was achieved. The inoculum was vortexed and 25 pL of the suspension was inoculated into the twelve wounds assigned for microbiology analysis. Serial dilutions of the suspension were plated onto culture media to quantify the exact concentration of viable organisms used for this experiment.

[0154] Treatment Regimen

[0155] Spray formulation wounds were sprayed with approximately 200 pL of test article from a spray bottle placed over the wounded area, which was then covered with a Tegaderm dressing (see FIGS. 2 to 4 for an example of this process).

[0156] The second treatment group had their wounds treated with 200 pL of silver sulfadiazine (SSD) each, which were then covered with a Tegaderm dressing (as shown in FIGS. 5 and 6).

[0157] Untreated control wounds were left without any treatment and covered only with a Tegaderm dressing (see FIG. 7).

[0158] All wounds were treated as described above daily for 10 days. All dressings were secured in place with tape and covered with Coban wrap (3M, St. Paul MN).

[0159] Clinical Observations

[0160] Wounds from each group were photographed and during each assessment time, the amount of erythema (redness) around the wounds were clinically scored.

[0161] Erythema - indicative of the amount of inflammation present*

[0162] * Score: 1 = absent, 2 = mild, 3 = moderate, 4 = marked, 5 = exuberant

[0163] All wounds inoculated with MRSA USA300, and non-inoculated wounds had erythema (mild score) observed on Day 1. By Day 4, only slight erythema was observed in all wounds. On Days 7 and 10, no erythema (absent score) was observed on all wounds. Recovery Methods

[0164] Microbiology Assessment

[0165] Four wounds were cultured on Day 10 after wounding and infection for enumeration of bacteria (in the assigned section for microbiology analysis). To recover bacteria from the wounds, a 6 mm punch biopsy was used in the wound area. The punch biopsies were taken from the center of the wound. Biopsies were weighed and immediately placed in 1 mL of All Purpose Neutralizing Solution. The sample was combined with an additional 4 mL of Neutralizing Solution and homogenized in a sterile homogenization tube. Serial dilutions were made from all culture samples and the extent of microbiological contamination assessed using the Spiral Plater System (Spiral Biotech, Norwood, MA). This system deposits a 50 mL aliquot of the scrub bacterial suspension over the surface of a rotating agar plate. Oxacillin Resistance Screening Agar (ORSAB) was used to isolate MRSA USA300 from the wounds. All plates were incubated aerobically overnight (24 - 48 hours) at 37°C, after which the number of viable colonies were counted. This method has been used for over 30 years to evaluate the antimicrobial efficacy of various topical agents and / or dressings.

[0166] Histological Assessment

[0167] For histological assessement, one incisional biopsy was taken on Days 4, 7 and 10 postwounding. Biopsies were obtained through the center of the wounds including normal adjacent skin on both sides. These specimens were placed in formalin then stained with hematoxylin and eosin (H&E). One section per block was analyzed. The specimens were then scanned on an Olympus VS 120 slide scanner and blindly evaluated for the following parameters to determine a potential treatment response:

[0168] Percent of wound epithelialized (%). Measurement of the length of the wound surface that has been covered with epithelium.

[0169] Epithelial thickness (cell layers inn). The epithelial thickness may vary from area to area within the biopsy. The thickness of the epithelium in pm was measured on five equal distance points from each other in the biopsy and averaged.

[0170] White cell infiltrate. Measured by the presence and amount of subepithelial mixed leukocytic infiltrates. Mean Score: 1 = absent, 2 = mild, 3 = moderate, 4 = marked, 5 = exuberant. Granulation Tissue Formation. The approximate amount of new granulation tissue formation (dermis) will be graded as follows: 0 = 0, 0.5 = 1-10%, 1 = 11-30%, 2 = 31-50%, 3 = 51-70%, 4 = 71-90%, 5= 91-100%

[0171] RESULTS

[0172] The data was combined, and analysis of the data was performed to determine the treatment response.

[0173] Microbiology Results

[0174] After counting the colonies, the data was tabulated and the Log of colony forming units / g (Log CFU / g) was determined with the mean of the Log (CFU / g) and standard deviation being calculated for each treatment.

[0175] On day 10, those wounds treated with spray formulation exhibited the lowest bacterial counts (4.84 ±0.32 Log CFU / g) in comparison to those treated with silver sulfadiazine and untreated control (5.64 ±0.19 and 6.32 ±0.10 Log CFU / g, respectively). Spray formulation also showed a bacterial reduction of 1.48 ±0.23 and 0.69 ±0. 10 Log CFU / g when compared to untreated control and silver sulfadiazine, respectively, representing 96.70% and 83.91% reduction of bacterial load reduction when compared to untreated control and silver sulfadiazine, respectively. Silver sulfadiazine treatment resulted in a reduction of 79.47% compared to untreated control wounds (FIG. 8).

[0176] Histology Results

[0177] The histological analyses were performed blindly without knowing the treatment of each group. The percentage of epithelialization and the other histological parameters were plotted against days after treatment. Since there were four samples for each treatment group per assessment time, mean and standard deviation were used for tabulation and graphical representation.

[0178] I. Percentage of re-epithelialization

[0179] The percentage of re-epithelialization represents the proportion of the wound area covered by newly formed epidermis with one or more layers of keratinocytes, which is a good index for the speed of keratinocyte migration and the first step of the re-epithelialization.

[0180] On day 4, wounds treated with the spray formulation exhibited a healing rate of approximately 21.9%, while wounds treated with silver sulfadiazine and untreated control had similar percentage of epithelialization 32.7% and 33.7%, respectively (FIG. 9). On day 7, wounds left untreated exhibited a re-epithelialization rate of 85.8%, while wounds treated with spray formulation had 82.5% of their wounds re-epithelialized and wounds treated with silver sulfadiazine showed the lowest re-epithelialization (80.4%). By day 10, all wounds were fully re-epithelialized (FIG. 9).

[0181] IL Epithelial thickness

[0182] The epithelial thickness was a measure of an average thickness of five points of newly formed epithelium. Epithelial thickness reflects the process of keratinocy te proliferation, differentiation, and epidermal maturation.

[0183] On Day 4, wounds treated with spray formulation and those wounds left untreated showed similar epithelial thickness (161.0 and 157.9 pm, respectively), whereas wounds treated with silver sulfadiazine were the thinnest on day 4 (134.0 pm) as shown in FIG. 10.

[0184] On days 7 and 10, the same trend was observed with silver sulfadiazine showing the thinnest epidermal thickness on both days (131.3 and 122.3 pm, respectively). On day 7, wounds treated with spray formulation had the thickest epidermis (157.8 pm), followed for untreated wounds (141.2 pm), with similar results observed with day 10 wounds (141.3 and 139.8 pm, respectively). It should be noted however, that for all treatments, epithelial thickness decreased from day 4 to day 10.

[0185] III. White cell infiltration

[0186] White cell infiltration (WCI) is used to access the inflammation reaction that could be a part of the normal process of wound repair, due to microbial infection, or the tissue reaction to foreign materials in the wound.

[0187] On day 4, wounds treated with silver sulfadiazine or left untreated exhibited the highest WCI scores at 1.19 and 0.96, respectively (FIG. 11), whereas on days 7 and 10, wounds treated with silver sulfadiazine and left untreated showed a higher WCI in comparison to the spray formulation treated wounds, which exhibited the lowest WCI on all assessment days (0.90, 0.43 and 0.39, respectively).

[0188] IV. Granulation tissue formation

[0189] The dermal reconstitution begins in about 3 to 4 days of injury with the hallmark of granulation tissue formation, which include new blood vessel formation (angiogenesis), and the accumulation of fibroblasts and collagen extracellular matrices. The granulation tissue formation measures the percent of wound bed filled with newly formed granulation tissue.

[0190] In all assessment days, all wounds had similar granulation tissue formation scores with values between 0.50 and 0.75 as shown in FIG. 12. For spray formulation the granulation score was 0.50 for all assessment days.

[0191] CONCLUSIONS

[0192] For the microbiology analysis, wounds treated with a spray formulation of the present teachings exhibited better efficacy in MRSA bacterial reduction when compared against a well established formulation (silver sulfadiazine) or untreated control in this study. The bacterial reduction was almost one and a half Log CFU / g less when compared to untreated wounds, representing 96.7% of reduction. In a head-to-head comparison with a known positive control (silver sulfadiazine), the spray formulation exhibited a 0.69 Log CFU / g bacterial reduction, reflecting a potential to reduce MRSA bioburden in infected burn wounds commonly detrimental to the healing process.

[0193] Moreover, histological results showed no detrimental effects for the spray formulation compared to controls. The spray formulation had a rapidly increase in epithelialization from day 4 to day 7 for more than 60%. Overall, the test spray formulation exhibited a promising antimicrobial effect without delaying the healing process.

[0194] Example 4. Investigation of Antimicrobial and Disinfectant Properties of Composite Material Solution

[0195] Bacteria used: Escherichia coli, Staphylococcus aureus, Acinetobacter baumannii

[0196] Escherichia coli: The most common gram-negative pathogenic bacteria. This bacterium is representative of all gram-negative bacteria. The fact that disinfectants have a significant effect on this bacterium indicates that these agents can be effective against all gram-negative bacteria.

[0197] Staphylococcus aureus: The most common gram-positive pathogenic bacteria. This bacterium is representative of gram-positive bacteria. The fact that disinfectants have a significant effect on this bacterium indicates that these agents can be effective against most gram-positive bacteria. Acinetobacter baumanniv. This bacterium is one of the most resistant and dangerous bacteria that cause hospital-acquired infections. These bacteria are resistant to most antibiotics and are also widely present in the environment. Infections with Acinetobacter are particularly important in hospitalized patients with immune system deficiencies.

[0198] Fungi used: Candida albicans, Aspergillus niger, Aspergillus flavus

[0199] The fungi used are among the most common fungi that cause infections in patients and are also widely present in the surrounding environment. For this reason, they are among the common infections in hospitalized patients, especially those with immune deficiencies.

[0200] Tests carried out:

[0201] Microbial contamination test of the sample, and evaluation of the surface antimicrobial properties on different surfaces (steel, ceramic, stone, and fabric) at different concentrations and times were carried out. The composite material formulation used in this example is that prepared in Example 1C.

[0202] Each test was repeated 6 times. Standard times for evaluating antimicrobial properties are usually 12 and 24 hours after the use of the substance. At the company's request, a 48-hour time was also added, and the requested amounts were considered in the tests.

[0203] General notes to consider:

[0204] In the discussion of surface disinfection, the type of microbe under investigation is not important, and the presence of any type of bacteria, fungi, parasites, and other microorganisms is an indication of the loss of antimicrobial properties.

[0205] In this regard, both pathogenic and non-pathogenic environmental bacteria are of interest, and the type of microbe does not require identification.

[0206] Positive control refers to bacteria that were added to the environment, but in the samples under investigation, it can be any type of environmental contamination.

[0207] Example 4a. Test for Internal Microbial Contamination of Composite Material Solution

[0208] In this test, the sterility of the mentioned samples is examined by inoculating them onto bacterial culture media and incubating them at various temperatures. This test is performed to ensure the absence of bacteria in biological and non-biological materials. Method:

[0209] One Cubic Centimeter (1 milliliter) of the tested samples is inoculated onto a nutrient-rich culture medium of TSB, and incubated for 24 hours at 37°C. Another sample is incubated for 48 hours at 24°C, the same temperature as the environment. After this period, the samples are compared to a standard sample for turbidity measurement to ensure microbial growth and sterility.

[0210] One Cubic Centimeter (1 milliliter) of the tested samples is inoculated onto a non-specific culture medium of Blood Agar, using a standard microbiology sampler and loop, in a lawn culture form and cultured for 24 hours at 37°C. Another sample is incubated for 48 hours at 24 °C, (room temperature).

[0211] Results:

[0212] • No turbidity was observed in the cultured samples in the TSB Environment.

[0213] • No colonies were observed on the Blood Agar inoculated with the samples.

[0214] • As a result, the composite material solution has sterile conditions with no bacterial contamination.

[0215] Example 4b. Internal Fungal Contamination Tests for the Composite Material Solution

[0216] In this test, the sterility of the mentioned samples is examined by inoculating them onto specific fungal culture media and incubating them at different temperatures. This test is performed to ensure the absence of fungi in biological and non-biological materials.

[0217] Method: for the purpose of this test, one Cubic Centimeter (1 milliliter) of sample is cultured on a fungal-specific culture medium and incubated in an incubator at a temperature of 37 degrees Celsius for 48 hours. Another sample is incubated at 24 °C (room temperature) in an incubator.

[0218] Result:

[0219] • From the perspective of fungal contamination, the composite material solution was completely sterile and no fungal colony growth was observed in the culture media. Example 4c. Analysis of the Antibacterial Properties of the Composite Material

[0220] Solution |Against Escherichia coli, Staphylococcus aureus, and Acinetobacter baumannii in Laboratory Culture Media.

[0221] Method:

[0222] In this method, a specific amount (100 microliters) of the solution under investigation is inoculated into wells created in Mueller-Hinton agar medium, which has been contaminated with the desired bacterial strain by lawn culture form culture. The culture media are then incubated in an incubator at 37°C for 24 hours, and the results are observed in the form of a lack of bacterial growth, indicated by a clear zone.

[0223] Result:

[0224] • This test was performed on Escherichia coli as a representative of gram-negative bacteria, Staphylococcus aureus as a representative of gram-positive bacteria, and Acinetobacter baumannii as a representative of environmental-resistant bacteria. All of these microorganisms were prevented from growing, and a significant clear zone was observed, indicating the antibacterial activity of the composite material solution .

[0225] Example 4d. Investigation of Antifungal Properties of the Composite Material Solution on Candida albicans, Aspergillus niger, Aspergillus flavus in Laboratory Culture Media

[0226] Method:

[0227] In this method, the desired amount (100 microliters) of the sample is poured onto sterile gas and placed on a specific culture medium for fungi that have been contaminated with Candida albicans, Aspergillus niger, and Aspergillus flavus. After 24 hours of incubation at 37°C and 24 °C, it is examined.

[0228] Result:

[0229] No fungal colonies grew in any of the fungal cultures on the area under the fabric soaked in the composite material solution. Example 4e. Investigation of Antimicrobial Properties of the Composite Material Solution by Spraying on the Surface of the Tile for Standard Amounts and Times

[0230] Method:

[0231] To examine the effect of the desired solution on disinfecting the environment (tiles), a positive control (tiles contaminated with gram-positive and gram-negative bacteria in the laboratory) and a negative control (tiles placed under ultraviolet radiation for several hours) were used. From each of the tiles, a sample was taken by sterile gas immersed in a rich liquid culture medium and cultured in a non-specific agar medium in a lawn culture form and incubated in an incubator at a temperature of 37°C and 24°C (ambient temperature). Positive and negative controls were also sampled and incubated like other samples and were examined after 24 and 48 hours.

[0232] Results:

[0233] Table 1

[0234] * Referring to Tables 1-7 in this Example, few colonies refers to the time when the disinfectant begins to lose its properties and the microorganisms start to colonize the surface, but not extensively, as there is still some effect of the substance on the surface.

[0235] The composite material solution at the highest concentration for at least 24 hours and at the lowest concentration for at least 12 hours can keep the surface of the tile sterile. Example 4f. Investigation of Antimicrobial Properties of the Composite Material Solution by Spraying on the Surface of the Stone for Standard Amounts and Times

[0236] Method:

[0237] To investigate the effect of the solution on disinfecting the environment (stone), positive control (stone contaminated with gram-positive and gram-negative bacteria in the laboratory) and negative control (stone placed under ultraviolet light for several hours) are used. A sterile cloth is soaked in the liquid and used to sample the surface of each stone. The samples are then cultured in a non-specific agar medium in a lawn culture form and incubated at 37°C and 24°C (ambient temperature). Positive and negative controls are also sampled and incubated like the other samples, and the results are examined after 24 and 48 hours.

[0238] Results:

[0239] Table 2

[0240] The composite material solution at the highest concentration for at least 24 hours and at the lowest concentration for at least 12 hours can keep the surface of the stone sterile.

[0241] Example 4g. Investigation of Antimicrobial Properties of the Composite Solution by Spraying on the Surface of the Ceramic for Standard Amounts and Times

[0242] Method:

[0243] To investigate the effect of the solution on disinfecting the environment (ceramic), a positive control (ceramic contaminated with gram-positive and gram-negative bacteria in the laboratory) and a negative control (ceramic exposed to ultraviolet radiation for several hours) are used. From each of the ceramics, a sample is taken by a sterile cloth soaked in a rich liquid culture medium and cultured in a non-selective medium in a in a lawn culture form and incubated at 37°C and 24°C (ambient temperature) in an incubator. Positive and negative controls are also taken and incubated like other samples. After 24 and 48 hours, they are examined.

[0244] Results:

[0245] Table 3

[0246] The composite material solution at the highest concentration for at least 24 hours and at the lowest concentration for at least 12 hours can keep the surface of the ceramic sterile.

[0247] Example 4h. Investigation of Antimicrobial Properties of the Composite Material Solution by Spraying on the Surface of Wood for Standard Amounts and Times

[0248] Method:

[0249] To investigate the effect of the solution on disinfecting the environment (Wood), a positive control (wood contaminated with gram-positive and gram-negative bacteria in the laboratory) and a negative control (wood exposed to ultraviolet radiation for several hours) are used. From each of the woods, a sample is taken by a sterile cloth soaked in a rich liquid culture medium and cultured in a non-selective medium in a in a lawn culture form and incubated at 37°C and 24°C (ambient temperature) in an incubator. Positive and negative controls are also taken and incubated like other samples. After 24 and 48 hours, they are examined.

[0250] Results:

[0251] Table 4

[0252] The composite material solution at the highest concentration for at least 24 hours and at the lowest concentration for at least 12 hours can keep the surface of the wood sterile.

[0253] Example 4i. Investigation of Antimicrobial Properties of the Composite Material Solution by Spraying on the Surface of Steel for Standard Amounts and Times

[0254] Method:

[0255] To investigate the effect of the solution on disinfecting the environment (Steel), a positive control (steel contaminated with gram-positive and gram-negative bacteria in the laboratory) and a negative control (steel exposed to ultraviolet radiation for several hours) are used. From each of the steels, a sample is taken by a sterile cloth soaked in a rich liquid culture medium and cultured in a non-selective medium in a lawn culture form and incubated at 37°C and 24°C (ambient temperature) in an incubator. Positive and negative controls are also taken and incubated like other samples. After 24 and 48 hours, they are examined.

[0256] Results:

[0257] Table 5

[0258] The composite material solution at the highest concentration for at least 48 hours and at the lowest concentration for at least 12 hours can keep the surface of the steel sterile.

[0259] Example 4j. Investigation of Antimicrobial Properties of the Composite Material Solution by Spraying on the Surface of the Fabric for Standard Amounts and Times

[0260] Method:

[0261] To investigate the effect of the solution on disinfecting the environment (Fabric), a positive control (fabric contaminated with gram-positive and gram-negative bacteria in the laboratory) and a negative control (fabric exposed to ultraviolet radiation for several hours) are used. From each of the fabrics, a sample is taken by a sterile swab soaked in a rich liquid culture medium and cultured in a non-selective medium in a lawn culture form and incubated at 37 °C and 24°C (ambient temperature) in an incubator. Positive and negative controls are also taken and incubated like other samples. After 24 and 48 hours, they are examined.

[0262] Results:

[0263] Table 6

[0264] The composite material solution at the highest concentration for at least 12 hours and at the lowest concentration for less than 12 hours can keep the surface of the fabric sterile.

[0265] Example 4k. Investigation of Antimicrobial Properties of the Composite Material Solution by Spraying on the Surface of the Nanofiber Fabric for Standard Amounts and Times

[0266] Method:

[0267] Two pieces of Nanofiber fabric (1 nr) immersed on solutions of 250 milliliters and 350 milliliters of the composite material solution in order, and placed them on the surface designated for the experiment. Then, at different times, samples were taken using sterile swabs from the surface covered with the fabric and cultured on non-specific blood agar media with a linear method. The cultured media were then incubated at 37°C and 24°C (ambient temperature) in an incubator. The area under UV radiation was used as a negative control, and the surface contaminated with gram-positive and gram-negative bacteria was used as a positive control. The results were examined and recorded after 24 hours.

[0268] Results:

[0269] Table 7

[0270] Using nano fiber fabric soaked in composite material solution at the highest concentration, for 24 hours and at the lowest concentration for 12 hours can keep the environment sterile. It seems that using nano fiber fabric is effective in preventing the rapid drying and evaporation of the solution. INCORPORATION BY REFERENCE

[0271] This application refers to various issued patents, published patent applications, journal articles, and other publications, all of which are incorporated herein by reference. If there is a conflict between any of the incorporated references and the instant specification, the specification shall control. In addition, any particular embodiment of the present teachings that falls within the prior art may be explicitly excluded from any one or more of the claims. Because such embodiments are deemed to be known to one of ordinary skill in the art, they may be excluded even if the exclusion is not set forth explicitly herein. Any particular embodiment of the disclosure can be excluded from any claim, for any reason, whether or not related to the existence of prior art.

[0272] EQUIVALENTS

[0273] The disclosure may be embodied in other specific forms without departing from the spirit or essential characteristics thereof. The foregoing embodiments are therefore to be considered in all respects illustrative rather than limiting the disclosure described herein. Scope of the disclosure is thus indicated by the appended claims rather than by the foregoing description, and all changes that come within the meaning and range of equivalency of the claims are intended to be embraced therein.

Claims

CLAIMSWhat is claimed is:

1. A composite material comprising TiCL coated Ag (TiCh / Ag) particles and TiCL coated Cu (TiCL / Cu) particles, wherein the TiOi / Ag particles and the TiCL / Cu particles are associated with a polymer in a vehicle.

2. The composite material of claim 1, further comprising TiCL coated Ag-Cu (TiCL / Ag- Cu) particles.

3. The composite material of claim 1 or 2, wherein the vehicle comprises water.

4. The composite material of any one of claims 1-3, wherein the composite material further comprises selenium.

5. The composite material of any of claims 1-4, wherein the polymer is polyvinylpyrrolidone, polyvinyl alcohol, polyethylene glycol, and combinations thereof.

6. The composite material of any one of claims 1-5, wherein the composite material further comprises chitosan.

7. The composite material of claim 6, wherein the weight ratio of the chitosan to the metals and metal oxides is in a range of about 0: 1 to about 10:1.

8. The composite material of claim 6 or 7, wherein the weight ratio of the chitosan to the polymer is in a range of about 0: 1 to about 7: 1.

9. The composite material of any one of claims 6-8, wherein the concentration of the chitosan is about 1 mg / L to about 2500 mg / L.

10. The composite material of any one of claims 1-9, wherein the concentration of the TiCL is about 50 mg / L to about 400 mg / L.

11. The composite material of any one of claims 1-10, wherein the concentration of the Ag is about 1 mg / L to about 60 mg / L.

12. The composite material of any one of claims 1-11, wherein the concentration of the Cu is about 1 mg / L to about 10 mg / L.CLAIMSWhat is claimed is:

1. A composite material comprising TiCL coated Ag (TiCh / Ag) particles and TiCL coated Cu (TiCL / Cu) particles, wherein the TiOi / Ag particles and the TiCL / Cu particles are associated with a polymer in a vehicle.

2. The composite material of claim 1, further comprising TiCL coated Ag-Cu (TiCL / Ag- Cu) particles.

3. The composite material of claim 1 or 2, wherein the vehicle comprises water.

4. The composite material of any one of claims 1-3, wherein the composite material further comprises selenium.

5. The composite material of any of claims 1-4, wherein the polymer is polyvinylpyrrolidone, polyvinyl alcohol, polyethylene glycol, and combinations thereof.

6. The composite material of any one of claims 1-5, wherein the composite material further comprises chitosan.

7. The composite material of claim 6, wherein the weight ratio of the chitosan to the metals and metal oxides is in a range of about 0: 1 to about 10:1.

8. The composite material of claim 6 or 7, wherein the weight ratio of the chitosan to the polymer is in a range of about 0: 1 to about 7: 1.

9. The composite material of any one of claims 6-8, wherein the concentration of the chitosan is about 1 mg / L to about 2500 mg / L.

10. The composite material of any one of claims 1-9, wherein the concentration of the TiCL is about 50 mg / L to about 400 mg / L.

11. The composite material of any one of claims 1-10, wherein the concentration of the Ag is about 1 mg / L to about 60 mg / L.

12. The composite material of any one of claims 1-11, wherein the concentration of the Cu is about 1 mg / L to about 10 mg / L.

13. The composite material of any one of claims 4-12, wherein the concentration of the Se is about 0.01 mg / L to about 6 mg / L.

14. The composite material of any one of claims 1-13, wherein the concentration of the polymer is about 300 mg / L to about 5000 mg / L.

15. The composite material of any one of claims 1-14, comprising about 93 % to about 99.9 % by weight of the vehicle based on the total weight of the composite material.

16. The composite material of any one of claims 1-15, wherein the composite material further comprises zinc oxide (ZnO).

17. The composite material of claim 16, wherein the concentration of the ZnO is about 0.001 mg / L to about 75 mg / L.

18. The composite material of any one of claims 1-17, wherein the composite material further comprises an acid.

19. The composite material of claim 18, wherein the acid comprises acetic acid, hydrochloric acid, or combinations thereof.

20. The composite material of claim 19, wherein the acid is acetic acid and the concentration of acetic acid is about 0.001% to about 1 %.

21. A composite material comprising: titanium dioxide coated silver particles; titanium dioxide coated copper particles; polyvinylpyrrolidone, and water.

22. A composite material comprising: titanium dioxide coated silver particles; titanium dioxide coated copper particles; selenium; polyvinylpyrrolidone, and water.

23. The composite material of claim 21 or 22, further comprising titanium dioxide coated silver-copper particles.

24. The composite material of any one of claims 21-23, further comprising one or more of zinc oxide, acetic acid, and chitosan.

25. The composite material of any one of claims 21-24, further comprising polyethylene glycol.

26. The composite material of any one of claims 1-25, wherein the composite material is in the form of a spray.

27. The composite material of any one of claims 1-25, wherein the composite material is positioned or mounted on a solid support.

28. A method of making a composite material comprising: mixing TiCh, AgNCh and Cu(NO3)*3H2O in a first vehicle to form a first mixture; mixing a polymer in a second vehicle to form a second mixture; mixing the first mixture with the second mixture to form a final mixture; and exposing the final mixture to a reducing agent until less than about 15% of the Ag is Ag+thereby forming a composite material.

29. The method of claim 28, wherein the reducing agent comprises ultraviolet radiation, microwaves, or a combination thereof.

30. The method of claim 28 or 29, wherein less than 10% by weight of the Ag is Ag+.

31. The method of any one of claims 28-30, wherein mixing to form the first mixture comprising mixing Na2SeC>4 with the TiCh, AgNCh and Cu(NO3)*3H2O.

32. A method of promoting wound healing in a subject in need thereof comprising contacting an effective amount of the composite material of any one of claims 1 -25 with a wound of the subject.

33. A method of claim 32, further comprising exposing the composite material to visible light, sunlight and / or UV light for a suitable time and under conditions suitable for the composite material to promote wound healing.

34. The method of claim 32 or 33, wherein the subject is a human.

35. The method of any one of claims 32-34, wherein the method comprises contacting fresh, unused composite material with the wound numerous times.

36. The method of claim 35, wherein numerous times comprises contacting fresh, unused photocatalytic composite material with the wound once per day, or once every other day, or once every three days.

37. The method of any one of claims 32-36, wherein the wound healing comprises treating a condition selected from the group consisting of first, second, or third degree burns; a compromise of the superficial layer, keratin layer, epithelial layer or deeper layers of skin; a disease that cause compromise of the superficial or deep layers of skin; skin colonization with any bacterial, microbial or fungal species; a compromise of superficial layers, internal epithelium, including superficial or deep layers of the gastrointestinal system; a compromise of internal epithelium or superficial and deeper layers of the anorectal system; a compromise of superficial and internal layers of the oral and dental mucosa; surgical debridement or intervention; pre- or post-surgical wound in any part of the body; an intervention or prevention of facial, scalp, oral, periportal, anal, peri-anal or any mucosal infection; a puncture; a rash; and an acneiform disorder.

38. The method of any one of claims 32-37, wherein contacting the composite material comprises topically applying the composite material, optionally, spraying the composite material, or applying the composite material mounted on a solid phase.

39. A method of treating or preventing an infection of a subject in need thereof comprising contacting an effective amount of the composite material of any one of claims 1 - 25 with an infection or an area of potential infection of the subject to treat or prevent the infection.

40. The method of claim 39, wherein the composite material is in the form of a mouth rinse.

41. A method of disinfecting a surface comprising contacting a surface with an effective amount of the composite material of any one of claims 1-25.5 42. The method of claim 41, wherein the surface is tile, wood, fabric, metal, stone, ceramic, or combinations thereof.

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