Treatment system for multiple types of microbial infections

Abstract

A treatment system for a microbial infection including a solution having a source of ions in the amount of about 25-225 mM, and a source of an oxidizing agent in the amount of about 25-225 mM, water, and a delivery device comprising polysaccharide(s) forming a three-dimensional polymeric biphasic network housing the solution.

Description

TREATMENT SYSTEM FOR MULTIPLE TYPES OF MICROBIAL INFECTIONSCROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of U.S. provisional application Serial No.63 / 527,930 filed July 20, 2023, and U.S. provisional application Serial No. 63 / 656,844 filed June 6, 2024, the disclosures of which are hereby incorporated in their entirety by reference herein.TECHNICAL FIELD

[0002] The present disclosure relates to a treatment or therapeutic system including a formulation for ionically charged chemical composition(s) and a delivery device as well as method of preparing and using the same.BACKGROUND

[0003] Microbial infections, including infections by bacteria, fungi, and / or viruses are a significant risk to humans and animals, and are a burden to healthcare systems worldwide. Many antimicrobial agents have been discovered or produced to combat these infections. Many of these agents, however, are subject to antimicrobial resistance. Additionally, these agents often elicit many unpleasant or even harmful side effects in both humans and animals. For example, antibiotics often cause digestive distress, partially due to their effect on the gut microbiome. Antibiotic use may also increase the incidence of fungal infections, e g., with Candida yeast, or infections with bacteria such as Clostridium difficile. Anti-fungal agents can be toxic due to their effect on cell membrane synthesis, a process that is largely similar between animal and fungal cells. Similarly, anti-viral agents often induce harmful side effects, including mitochondrial, neurologic, and hematopoietic toxicity.SUMMARY

[0004] In at least one embodiment, a treatment system for a microbial infection is disclosed. The treatment system may comprise a solution including a source of ions in the amount of about 25-225 mM, a source of an oxidizing agent in the amount of about 25-225 mM, water,and a delivery device comprising polysaccharide(s) forming a three-dimensional polymeric biphasic network housing the solution. The polysaccharide(s) may include polyvinyl alcohol. The network may form a body with a centrally-located aperture. The body may have a raised portion adjacent the centrally-located aperture. The body may include a second aperture running from an edge of the body to the centrally-located aperture. The source of ions may include zinc chloride (ZnCl2). The oxidizing agent may include sodium chlorite (NaClO2). The delivery device may be a hydrogel with a uniform thickness.

[0005] In another embodiment, a treatment system is disclosed. The treatment system may comprise an aqueous solution including a source of ions in the amount of about 25-225 mM, a source of an oxidizing agent in the amount of about 25-225 mM, and a thermally reversible delivery device housing the solution. The delivery device may have a first state of being a liquid at a first temperature range and a second state of being a gel at a second temperature range. The second temperature range may be different than the first temperature range. The second temperature range may be higher than the first temperature range. The delivery device in the second state may mold to a shape of a surface it is applied to. The thermally-reversible delivery device may be a hydrogel. The delivery device may be a dressing. The thermally-reversible delivery device may include one or more poloxamers.

[0006] In yet another embodiment, a treatment system is disclosed. The treatment system may comprise an aqueous solution comprising about 25-225 mM of a source of Zn2+, Ag+, or Cu2+ ions, about 25-225 mM of a source of an oxidizing agent forming CIO2' ions, about 0.5-10 mM of a succinate, and about 0-5 mM ascorbic acid. The aqueous solution may be configured to increase a redox potential at a target site. The treatment system may also include a delivery device housing the solution. The delivery device may be a hydrogel. Ascorbic acid may be present. The aqueous solution may additionally include chlorhexidine gluconate. The aqueous solution may additionally include hyaluronic acid. The aqueous solution may additionally include one or more poloxamers.BRIEF DESCRIPTION OF THE DRAWINGS

[0007] FIGURE 1A is a top view of a non-limiting example of a hydrogel including formula according to one or more embodiments disclosed herein;

[0008] FIGURE IB is a top view of an alternative non-limiting example of a hydrogel according to the disclosure;

[0009] FIGURES 1C, ID, IE show additional non-limiting example hydrogel structures according to the disclosure;

[0010] FIGURE IF shows a perspective view of a non-limiting example of a hydrogel disclosed herein cooperating with a medical instrument;

[0011] FIGURE 2 shows a non-limiting example of a dressing impregnated with a formula disclosed herein; and

[0012] FIGURES 3-10 shows formulas and test results for bacterial inhibition on hydrogels, according to one or more embodiments disclosed herein, of Examples 11-18.DETAILED DESCRIPTION

[0013] Reference will now be made in detail to presently preferred compositions, embodiments and methods of the present disclosure, which constitute the best modes of practicing the disclosure presently known to the inventors. The Figures are not necessarily to scale. However, it is to be understood that the disclosed embodiments are merely exemplary of the disclosure that may be embodied in various and alternative forms. Therefore, specific details disclosed herein are not to be interpreted as limiting, but merely as a representative basis for any aspect of the disclosure and / or as a representative basis for teaching one skilled in the art to variously employ the present disclosure.

[0014] Except in the examples, or where otherwise expressly indicated, all numerical quantities in this description indicating amounts of material or conditions of reaction and / or use are to be understood as modified by the word "about" in describing the broadest scope of the disclosure. Practice within the numerical limits stated is generally preferred. Also, unless expressly stated to the contrary: the description of a group or class of materials as suitable or preferred for a given purpose in connection with the disclosure implies that mixtures of any two or more of the members of the group or class are equally suitable or preferred; description of constituents inchemical terms refers to the constituents at the time of addition to any combination specified in the description, and does not necessarily preclude chemical interactions among the constituents of a mixture once mixed; the first definition of an acronym or other abbreviation applies to all subsequent uses herein of the same abbreviation and applies mutatis mutandis to normal grammatical variations of the initially defined abbreviation; and, unless expressly stated to the contrary, measurement of a property is determined by the same technique as previously or later referenced for the same property.

[0015] It must also be noted that, as used in the specification and the appended claims, the singular form "a," "an," and "the" comprise plural referents unless the context clearly indicates otherwise. For example, reference to a component in the singular is intended to comprise a plurality of components.

[0016] As used herein, the term “about” means that the amount or value in question may be the specific value designated or some other value in its neighborhood. Generally, the term “about” denoting a certain value is intended to denote a range within + / - 5% of the value. As one example, the phrase “about 100” denotes a range of 100+ / - 5, i.e. the range from 95 to 105. Generally, when the term “about” is used, it can be expected that similar results or effects according to the disclosure can be obtained within a range of + / - 5% of the indicated value.

[0017] As used herein, the term “and / or” means that either all or only one of the elements of said group may be present. For example, “A and / or B” shall mean “only A, or only B, or both A and B”. In the case of “only A”, the term also covers the possibility that B is absent, i.e. “only A, but not B”.

[0018] It is also to be understood that this disclosure is not limited to the specific embodiments and methods described below, as specific components and / or conditions may, of course, vary. Furthermore, the terminology used herein is used only for the purpose of describing particular embodiments of the present disclosure and is not intended to be limiting in any way.

[0019] The term “comprising” is synonymous with “including,” “having,” “containing,” or “characterized by.” These terms are inclusive and open-ended and do not exclude additional, unrecited elements or method steps. The phrase “consisting of’ excludes any element, step, oringredient not specified in the claim. When this phrase appears in a clause of the body of a claim, rather than immediately following the preamble, it limits only the element set forth in that clause; other elements are not excluded from the claim as a whole. The phrase “consisting essentially of’ limits the scope of a claim to the specified materials or steps, plus those that do not materially affect the basic and novel characteristic(s) of the claimed subject matter. The phrase “composed of’ means “including” or “consisting of.” Typically, this phrase is used to denote that an object is formed from a material. With respect to the terms “comprising,” “consisting of,” and “consisting essentially of,” where one of these three terms is used herein, the presently disclosed and claimed subject matter can include the use of either of the other two terms.

[0020] The term “one or more” means “at least one” and the term “at least one” means “one or more.” The terms “one or more” and “at least one” include “plurality” as a subset.

[0021] The term “substantially,” “generally,” or “about” may be used herein to describe disclosed or claimed embodiments. The term “substantially” may modify a value or relative characteristic disclosed or claimed in the present disclosure. In such instances, “substantially” may signify that the value or relative characteristic it modifies is within ± 0%, 0.1%, 0.5%, 1%, 2%, 3%, 4%, 5% or 10% of the value or relative characteristic.

[0022] It should also be appreciated that integer ranges explicitly include all intervening integers. For example, the integer range 1-10 explicitly includes 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10. Similarly, the range 1 to 100 includes 1, 2, 3, 4 . . . 97, 98, 99, 100. Similarly, when any range is called for, intervening numbers that are increments of the difference between the upper limit and the lower limit divided by 10 can be taken as alternative upper or lower limits. For example, if the range is 1.1. to 2.1 the following numbers 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, and 2.0 can be selected as lower or upper limits.

[0023] In the examples set forth herein, concentrations, temperature, and reaction conditions (e.g., pressure, pH, flow rates, etc.) can be practiced with plus or minus 50 percent of the values indicated rounded to or truncated to two significant figures of the value provided in the examples. In a refinement, concentrations, temperature, and reaction conditions (e.g., pressure, pH, flow rates, etc.) can be practiced with plus or minus 30 percent of the values indicated rounded to or truncated to two significant figures of the value provided in the examples. In anotherrefinement, concentrations, temperature, and reaction conditions (e.g., pressure, pH, flow rates, etc.) can be practiced with plus or minus 10 percent of the values indicated rounded to or truncated to two significant figures of the value provided in the examples.

[0024] Throughout this application, where publications are referenced, the disclosures of these publications in their entireties are hereby incorporated by reference into this application to more fully describe the state of the art to which this disclosure pertains.

[0025] The following examples illustrate the various embodiments of the present disclosure. Those skilled in the art will recognize many variations that are within the spirit of the present disclosure and scope of the claims.

[0026] Healthcare associated infections, including urinary tract infections, surgical site infections, bloodstream infections, pneumonia, and other similar infections are a significant cause of morbidity and mortality in the United States. These infections also contribute substantially to increased healthcare costs.

[0027] Antibiotics were used for thousands of years to treat infections even before people realized that certain infections were caused by bacteria. The discovery in 1909 by Paul Ehrlich that the chemical arsphenamine (Salvarsan) effectively treated syphilis led to this compound serving as the first modem antibiotic. The term antibiotic, however, was not commonly used until nearly 30 years later after Alexander Fleming discovered penicillin. By 1941 , penicillin had been adapted from the laboratory to become a life-saving clinical treatment, due in part to the large-scale production of penicillin led by scientists and engineers during World War II. Penicillin also served as a replacement for the sulfa drugs that had previously been used as the first line of defense against bacterial infections. At this point, resistance to sulfa drugs had already been discovered. Penicillin was thus a welcomed defense to this deadly resistance.

[0028] Penicillin resistance was discovered almost simultaneously with the first mass production of the drug for clinical use. Fifteen years after a Nobel prize for the work establishing penicillin as an accessible clinical treatment was awarded, Methicillin-resistant Staphylococcus aureus (MRSA) was discovered. Since then, numerous additional multidrug-resistant bacteria have been discovered, including Vancomycin-resistant Enterococcus (VRE) and Vancomycin-resistant Staphylococcus aureus (VRSA). According to the Centers for Disease Control (CDC), multidrug-resistant bacteria cause 2.8 million infections and 35,000 deaths in the United States each year, and 5 million worldwide.

[0029] In addition to the existence of multidrug-resistant bacteria, antibiotics have other limitations. For example, antibiotics may cause numerous side effects including allergic reactions, gastrointestinal stress, physiologically harmful changes to the gut microbiome, weakening of one’s immune system, secondary infections with either yeast or bacteria that are resistant to the antibiotic, and the like. Antibiotics also do not work instantly, often taking up to 48 hours to produce measurable relief from the pathology and / or symptoms of an infection. This lag can lead to significant morbidity or even death. Additionally, antibiotics tend to be specific to certain types of bacteria.

[0030] Resistance to anti-fungal drugs has also been discovered. Candida auris, which is a multidrug-resistant pathogenic yeast, has recently emerged as a global health threat causing often fatal nosocomial infections in hospitals worldwide. Given that currently only three types of antifungal drugs exist, anti-fungal drug resistance is a serious threat. Similar to antibiotics, anti-fungal drugs may cause allergic reactions and gastrointestinal distress. They may also stress the liver and kidneys. Additionally, anti-fungal drugs may take weeks or months to reduce the level of the pathogenic fungus which may increase the incidence of side effects, or possibly lead to significant morbidity or even death.

[0031] Resistance to anti-viral drugs has also been observed, with acyclovir-resistant HSV being discovered in 1982. The most common mechanism for anti-viral drug resistance is genetic mutations in the viral DNA or RNA. While anti-viral resistant strains have been most commonly identified in immunocompromised patients, the rise of resistant strains may occur in animal reservoirs as well. For example, anti-virals consumed by humans may be excreted into wastewater. Animals coming into contact with such wastewater may develop resistant virus infections through selective mutation that occurs when the viruses come into sustained contact with the anti-virals. In addition to promoting resistance, anti-virals may also cause side effects similar to antibiotics and anti-fungal drugs, and take on the order of days or weeks to reduce infection, possibly increasingmorbidity or mortality. Additionally, antiviral medications tend to be specific to a specific virus, strain, type, or species.

[0032] Hence, there is an urgent need to develop alternate antimicrobial treatments that are timely and effective at preventing, reducing, and / or eliminating a broad spectrum of infections or the symptoms thereof without inducing antimicrobial resistance, or being susceptible to resistance, or causing undue side effects to a human or animal patient.

[0033] In one or more embodiments, a formulation, formula, composition, or solution is disclosed herein. The formulation may be applied to any site on or within a human or animal body, including but not limited to the skin, surgical sites, body cavities, organs, tissues, abscesses, and acute or chronically occurring wounds, bum wounds, graft sites (e.g., skin grafts). The formulation may be used to treat infections of any kind, such as, but not limited to, infections of the skin.

[0034] The formulation may be used to prevent, control, reduce, treat, and / or remove microorganisms, including gram positive and gram negative bacteria, aerobic and anaerobic bacteria, viruses, fungi, or any other organisms / microorganisms having infectious properties. The formulation may be a treatment or therapeutic formulation or solution.

[0035] The formulation may be a solution, liquid, solid, gel, or aerosol depending on the type of treatment site, type of delivery device, and components included in the formulation. For example, as will be discussed below, the formulation may be dispensed via a nebulizer or atomizer as a mist such that any solid components may be minimized. In another non-limiting embodiment, the formulation may be structured as a hydrogel including components enabling gelation. Other examples are described below.

[0036] The formulation may include a solution or a chemical mixture where one substance dissolves in another. The solution may be an aqueous solution. The solution may include a source of ions; a source of an oxidizing agent; and water. The solution may optionally include a buffer, a degradation inhibitor, a source of Vitamin C, a surfactant, or a combination thereof. The ions may be metal ions. The ions may be Zn2+, Ag+, Cu2+, Au+, or the like. The oxidizing agent may be, for example, sodium chlorite (NaCIO2). The buffer may include, but is not limited to, succinate.

[0037] The formulation may be formed by providing a first fluid having, comprising, consisting of, or consisting essentially of a source of ions; and by providing a second fluid having, comprising, consisting of, or consisting essentially of the oxidizing agent. The first fluid may include a source of Vitamin C. The first fluid may include a buffer and / or a surfactant. The second fluid may include a buffer and / or a surfactant. Upon mixing the first and second fluids together, the formula includes an ionically charged fluid with an oxidation-reduction potential higher than the oxidation-reduction potential at an infection site. The ionically charged fluid increases the antimicrobial activity at the infection site upon application. The solution may include the first and second fluid mixed together.

[0038] The solution providing the source of ions may be a solution forming Zn2+, Ag+, or Cu2+ions. The solution providing the source of ions, or mixed with such solution, may have preferred solubility for the specific ion selected, as well as provide an effective concentration of ions without permitting the redox potential to fall to unwanted levels. In one or more embodiments, the solution providing the source of ions may include salts of a weak acid to provide the free available ions in the charged compound. In at least one embodiment, the weak acid is soluble in water. Examples of zinc compounds that form solutions with free available zinc ions include, but are not limited to, zinc chloride, zinc acetate, zinc lactate, zinc salicylate, zinc sulfate, zinc nitrate, zinc stearate, zinc gluconate, zinc ammonium sulfate, zinc chromate, zinc citrate, zinc dithionate, zinc fluorosilicate, zinc tartrate, zinc formate, zinc iodide, zinc phenol sulfonate, zinc succinate, zinc glycerophosphate, or other zinc halides. Examples of copper compounds that form solutions with free available copper ions include, but are not limited to copper acetate, copper formate, copper citrate, copper lactate, copper oxalate, copper propionate, copper benzoate, copper succinate, copper malonate, copper stearate, copper gluconate, copper ammonium sulfate, copper chromate, copper dithionate, copper fluorosilicate, copper tartrate, copper iodide, copper nitrate, copper phenol sulfonate, copper salicylate, copper sulfate, copper glycerophosphate, or other copper halides. Examples of silver compounds that form solutions with free available silver ions include, but are not limited to silver sulfate, silver oxide, silver acetate, silver nitrate, silver citrate, silver chloride, silver lactate, silver phosphate, silver stearate, silver thiocyanate, silver saccharinate, silver anthranilate, silver carbonate, or other silver halides.

[0039] The solution providing the source of an oxidizing agent may be a solution forming CIO2- ions. These CIO2- ions may oxidize the metal ions provided by the solution providing the source of ions, charging them, and thereby increasing the concentration of charged ions. For example, upon mixing the first and second fluids together, the chlorite from the solution providing the oxidizing agent may oxidize the zinc from the solution providing the source of ions, such that the concentration of Zn2+ions in the mixed solution increases.

[0040] The metabolism and growth of microorganisms at sites of infections, including but not limited to wounds and surgical sites (target sites), may increase when the oxidation-reduction (redox) potential (Eh) of the infection site is decreased. A decrease in oxygen at the infection site may decrease the redox potential of that site. Other changes in the biological and chemical properties of the infection site may also lead to a reduction in the redox potential of that site. By increasing the redox potential at the infection site, the metabolism and growth of harmful microorganisms, including but not limited to, bacteria, fungi or viruses, may be decreased, thus preventing, minimizing, and / or curing an infection.

[0041] In addition to ions and an oxidizing agent, the formula may include a Vitamin C component. The Vitamin C may be any form of Vitamin C such as, but not limited to, ascorbic acid, ascorbates such as mineral ascorbates, flavonoids, or mixtures or variations thereof having antioxidant properties. The Vitamin C component may have wound healing properties in addition to the antioxidant properties. In some instances, the Vitamin C component may have antimicrobial properties. The Vitamin C component may be included in an effective concentration, as determined by the type of Vitamin C component and the solutions provided. For example, the Vitamin C component may be about 0 mM to 5 mM, about 1 to 4 mM, or about 2 to 3 mM concentration of an ionically charged zinc solution. Additionally, in some embodiments, the formula may include additional vitamins, including, but not limited to, Vitamin A, Vitamin D, Vitamin E, Vitamin Bi- 6, Vitamin B12, and others and mixtures thereof. For example, B Vitamins may provide prophylactic properties against potential host tissue toxicity and / or oxidative damage of components in the formula.

[0042] The formula may also include chlorhexidine gluconate to prevent biofilm formation. The chlorhexidine gluconate may be provided in any effective amount in the formula.For example, the chlorhexidine gluconate may be about 0.01 to 2, 0.05 to 1.8, or 0.1 to 0.5% of the charged biofilm formula by volume. For example, the chlorhexidine gluconate may be about 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.1, 0.15, 0.2, 0.25, 0.3, 0.35, 0.4, 0.45, 0.5, 0.55, 0.6, 0.65, 0.7, 0.75, 0.8, 0.85, 0.9, 0.95, 1.0, 1.15, 1.2, 1.25, 1.3, 1.35, 1.4, 1.45, 1.5, 1.55, 1.6, 1.65, 1.7, 1.75, 1.8, 1.85, 1.9, 1.95, or 2 vol.% of the charged biofilm formula. The formula may alternatively or additionally include cetylpyridinium chloride, an antiseptic to kill bacteria and microorganisms in tissue and wounds. The addition of chlorohexidine gluconate and / or the cetylpyridinium chloride provides additional antimicrobial properties to the formula.

[0043] Furthermore, the formula may include at least one antibiotic agent, antiviral agent, antifungal agent, other antimicrobial agent, or combinations thereof. The antibiotic agent, antiviral agent, antifungal agent, other antimicrobial agent, or combinations thereof may be provided in a separate solution to be added to the formula. The antibiotic agent, antiviral agent, antifungal agent, other antimicrobial agent, or combinations thereof, may be selected specifically for the type of wound, surgical site, infection, or tissue to be irrigated / treated.

[0044] The formula may also include a protein such as a transforming growth factor beta (TGF-β) protein to promote wound healing. The TGF-β additive may be any of three isoforms ofTGF-β (β1, β2, β3) or a combination thereof.

[0045] The formula may include a peptide such as a proinsulin C-peptide to decrease inflammation.

[0046] The formula may include a pain-relieving agent including but not limited to an anesthetic, or analgesic, and / or combinations thereof. Non-limiting examples of anesthetics include bupivacaine hydrochloride, lidocaine, benzocaine, prilocaine, and the like. Non-limiting examples of analgesics include acetaminophen, ibuprofen, aspirin, naproxen, naproxen sodium, diclofenac, celecoxib, ketorolac, and the like.

[0047] The ions present in the solution providing the source of ions may be provided by ZnCl2solution. The concentration of the ZnCl2in the solution may be at least about 25 mM to about 225 mM. The concentration of the ZnCl2in the solution may be at least about, at most about, or about 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 115, 120, 125,130, 135, 140, 145, 150, 155, 160, 165, 170, 175, 180, 185, 190, 195, 200, 205, 210, 215, 220, or 225 mM. In a non-limiting example, the concentration of ZnCl2in the solution providing the source of ions may be about, at least about, or at most about 25 mM. In another non-limiting example, the concentration of ZnCl2in the solution providing the source of ions may be about, at least about, or at most about 50 mM. The pH of the solution providing the source of ions may be acidic, at about 4-6. In a non-limiting example, the pH of the solution providing the source of the ions may be about 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, or 7.

[0048] The Eh increasing component present in the solution providing the oxidizing agent may be provided by NaCIO2(sodium chlorite) solution. The concentration of NaCIO2in the solution providing the oxidizing agent may be at least about 25 mM to about 225 mM. The concentration of the NaCIO2in the solution may be at least about, at most about, or about 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 115, 120, 125, 130, 135, 140, 145, 150, 155, 160, 165, 170, 175, 180, 185, 190, 195, 200, 205, 210, 215, 220, or 225 mM. In a nonlimiting example, the concentration of NaCIO2may be at least about 25 mM. In another nonlimiting example, the concentration of NaCIO2is at least about 50 mM.

[0049] The ratio of the source of ions to the Eh increasing component may be about 1:1 such as about 50:50 mM, 100:100 mM, or 200:200 mM. Other ratios are also contemplated, for example 1 :2, 1 :3, or 1 :4 for the source of ions to the Eh increasing component or vice versa.

[0050] The concentration of ascorbic acid in the formula may be about 0.1 mM to about 5 mM. The concentration of ascorbic acid in the formula may be at least about, at most about, about 0, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, or 5.0 mM. It was unexpectedly discovered that ascorbic acid concentrations greater than 5 mM inhibit the effectiveness of the formula in eradicating planktonic bacteria. Multiple formulations containing varying amounts of ascorbic acid are contemplated. The varied levels of ascorbic acid from one formulation versus another provide differing levels of effectiveness against planktonic versus biofilm bacteria. Additional concentrations of ascorbic acid are contemplated based on the effectiveness of formulations containing such concentrations against other microbial organisms including, but not limited to, fungi and viruses.

[0051] Ascorbic acid may be absent such that that formulation is free of ascorbic acid. Higher concentrations of ascorbic acid may inhibit the prevention, reduction, speed, or elimination of free-floating planktonic bacteria. Ascorbic acid may aid in the prevention, reduction, and / or elimination of biofilm bacteria. It was observed that low concentrations of ascorbic acid were effective against biofilm bacteria while still remaining effective against planktonic bacteria as well. For example, ascorbic acid at a concentration of around 3 mM or less were effective against both biofilm and planktonic bacteria.

[0052] The formula may include additional components. The additional components may be part of the formula and / or part of the delivery device for the formula, as is discussed below. The one or more additional components may be part of both — the formula and the delivery device. The one or more components may include sodium chloride (NaCl), succinate or succinic acid, polysaccharide, alcohol, acid, starch, glycerin, hyaluronic acid, polyvinyl alcohol, poloxamer(s), or a combination thereof. The formula may be free of NaCl, succinate or succinic acid, alcohol, acid, polysaccharide, starch, glycerin, hyaluronic acid, polyvinyl alcohol, poloxamer(s), or a combination thereof.

[0053] NaCl may be included in the amount of about 0-50, 5-40, or 10-30 mM. The amount of sodium chloride may be about, at least about, or at most about 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, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 50 mM, or any range including any two of the numerals disclosed herein.

[0054] Succinate or succinic acid may be included in the amount of 0-25, 1-20, or 2-15 mM. Succinate or succinic acid may be included in the amount of about, at least about, or at most about 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 mM, or any range including any two of the numerals disclosed herein.

[0055] The formula / delivery device may include alcohol such as polyvinyl alcohol (PVA) at 0-30, 5-25, 8-20 wt. or vol.%, based on the weight of the formulation. PVA may be included in the amount of about, at least about, or at most about 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, or 30 wt. or vol.%, or any range including any two of the numerals disclosed herein.

[0056] The formulation / delivery device may include hyaluronic acid (HA) at about 0-5, 0.1-4, or 0.5-3 wt. or vol.%, based on the total weight or volume of the formulation. HA may be included in the amount of about, at least about, or at most about 0, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, or 5.0 wt. or vol.%, based on the total weight or volume of the formulation, or any range including any two of the numerals disclosed herein.

[0057] The formulation / delivery device may include a starch, and / or glycerin at about 1- 30, 2-25, or 3-20 wt. or vol.% each, based on the total weight or volume of the formulation. The starch and / or glycerin may be included in the amount of about, at least about, or at most about 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 wt. or vol.%, based on the total weight or volume of the formulation, or any range including any two of the numerals disclosed herein.

[0058] The formulation / delivery device may include one or more poloxamers. Example poloxamers may include Poloxamer 188, Poloxamer 407, and / or others. The amount of poloxamers in the formulation, when present, may be about 1-40 wt. or vol.%, based on the total weight or volume of the formulation. The one or more poloxamers may be present in the amount of about, at least about, or at most about 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, 40 wt.%, based on the total weight or volume of the formulation, or any range including any two of the numerals disclosed herein. Non-limiting example amounts of poloxamers may be 4, 20, 23, 29 wt. or vol.%.

[0059] The formula disclosed herein may be used at various types of infection sites to increase the redox potential to decrease metabolism and growth of bacteria in the wound or tissue, and also to decrease or eradicate the formation of biofilms.

[0060] The contemplated formulations or formulas are configured to be effective within minutes such that the formula may be used in surgical situations. The relatively fast speed of effectiveness of the formula may additionally decrease the opportunity for side effects in a patient. The speed also aids in the effectiveness of the formula as a prophylactic to be used in pre-surgical settings to prevent microbial infections at the surgical site.

[0061] The contemplated formulations may be configured to remain effective for days to weeks, and to gradually elute active formula solution to provide long-lasting antimicrobial effects.

[0062] A treatment system or therapeutic system or healing system utilizing the formula disclosed herein may include one or more components. The first component may be the formula / formulation / composition disclosed herein. The second component may include a delivery vehicle or device for the formula. The delivery device may include a hydrogel, dressing, colloidal gel, intravenous device, intra-articular injection, catheter, nebulizer, atomizer, mister, or other aerosol dispensing device, dispensing device, fabric such as wet wipe, the like, or a combination thereof. The delivery device may also be a container such as a squeeze tube, packet to be cut to remove the solution, or the like such that the solution may be directly applied onto the target site.

[0063] A hydrogel is a three-dimensional hydrophilic polymeric biphasic network arranged to absorb and retain one or more fluids without the structural network being dissolved. Hydrogels may deliver fluids, drugs, and biological molecules to delivery sites in human and animal bodies and tissues. The hydrogels designed to deliver the formula disclosed herein may be of a suitable composition, including but not limited to polysaccharide-based hydrogels that include chitosan, cellulose, dextran, starch, hyaluronic acid (HA) and the like; protein-based hydrogels that include collagen, gelatin, albumin and the like; poloxamers; poly(ethylene imine)- poly(ethylene glycol) methacrylate (PEI-PEGMA); polyvinyl-alcohol (PVA); or other such suitable materials or combinations thereof. Additionally, injectable hydrogel microspheres are contemplated as an example of a suitable delivery mechanism for the formula.

[0064] The hydrogel may house the solution disclosed herein. Upon placement on the body, the hydrogel may start releasing the solution at a rate. After a predetermined amount of time, the solution may be released into the body such that the hydrogel remains and the solution is all absorbed by the body. Subsequently, the hydrogel sans the solution may be removed from the target site.

[0065] The hydrogels used to deliver the formula disclosed herein may be used in conjunction with one or more percutaneous devices, including but not limited to, catheters, longterm implants, fixator pins, the like, or a combination thereof. Hydrogels used to deliver the formula to the site of a percutaneous pin may be of various shapes, sizes, and configurationsincluding, but not limited to, donut-shaped, discoid, cylindrical, semi-cylindrical, or the like. The hydrogel may be configured as a gasket seal around the entrance to the wound through which the percutaneous pin is inserted. The hydrogel may both seal the entrance to the wound around the pin, and deliver the formula to the wound area, thereby reducing microbial colonization and / or growth.

[0066] The hydrogel may include a void, opening, aperture, or through-hole in the center of its network. The diameter of the hole may correspond to a diameter of a percutaneous pin or another device being inserted through the hole. The fit of the device within the hole may be tight such that a gasket-like seal forms around the inserted device. The diameter may be about 1-10, 1.5-8, or 2-7 mm. The diameter may be about, at least about, or at most about 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 mm.

[0067] The thickness of the hydrogel may be uniform, non-uniform, regular, irregular. The hydrogel body may include one or more portions with different thickness than the body thickness. The different thickness may be greater or smaller than the body thickness. A non-limiting example of such portion may be a raised portion adjacent to a through-hole. Another example portion may include a portion which may be penetrated to become a through-hole. The hydrogel body may include one or more such portions. The hydrogel body may thus be adjustable to one or more devices requiring different though-hole locations. The raised portion may be configured to cooperate with the wound, for example by embedding in the wound.

[0068] A non-limiting example of a hydrogel for delivery of the formula disclosed herein the shown in Figs. 1 A, IB. The hydrogel 20 has a generally circular body 22 with a through-hole 24. The hydrogel body 22 also include a secondary void 26 running from an edge or perimeter 28 of the body 22 towards the center 30 with the through-hole 24. The hydrogel body 22 also includes a raised portion 25 adjacent the though-hole 24. The raised portion 25 may be a knob or an elevation. The raised portion 25 may provide a tight fit between a device (not depicted) to be inserted through the hole 24 and potentially the wound.

[0069] An alternative non-limiting example of a hydrogel disclosed herein is shown in Fig. IB. The hydrogel 20 includes a through-hole 24 and indentations 32. The indentations may bethrough-holes 24 as well or portions with a different thickness than the hydrogel body 22. The indentations may be regular or irregular.

[0070] Figs. 1C, ID, and IE shows alternative non-limiting embodiments of the hydrogel disclosed herein.

[0071] Fig. IF shows a non-limiting example of the hydrogel 20 applied around a medical device 50 such as a percutaneous pin. The hydrogel 20 may be provided around the device 50. Upon placement of the medical device 50 in a target site, the hydrogel 20 may form a gasket.

[0072] The delivery device of the solution may be thermally reversible or temperature dependent. A thermally reversible delivery device may be configured to transition from a liquid to a solid (gel) phase and back to the liquid phase depending on the temperature of the target site, environment, or both. The thermally reversible delivery device may include one or more compounds which are temperature-dependent, rending the delivery device a temperaturedependent solution / liquid / gel.

[0073] The thermo-reversible properties of the delivery device may be conferred by a poloxamer. Poloxamers are triblock copolymers. They are composed of a central block of polypropylene oxide with a block of polyethyleneoxide on either side. Poloxamers exist in a liquid phase at low temperatures and undergo gelation above a lower critical gelation temperature. Therefore, a poloxamer-based delivery device may be administered in a liquid form at room temperature via a dropper, syringe, or another type of applicator. Upon contact with a surface such as a human or animal tissue having a temperature higher than the lower critical gelation temperature, the hydrogel will undergo gelation.

[0074] The thermally reversible delivery device may be applied onto a target site such as a wound as a liquid. Upon contact with the target site— a body part— having a different temperature, the delivery device such as a hydrogel may form a gel / solid in the shape of the wound or tissue. The delivery device may thus mold into the shape of the body part which is being treated.

[0075] The thermally reversible gel may contain a concentration of one or more poloxamers that confer a lower critical gelation temperature that is higher than the temperature ina location such as a typical operating room, physician’s office, or average home, for example. The lower critical temperature of the thermally reversible delivery device may be such that the delivery device will undergo gelation upon contact with a wound site in human skin or tissue.

[0076] The thermally reversible delivery device may have at least a first state and a second state. The first state is a state in which the delivery device is a liquid. The first state corresponds to a first temperature range which is lower than the critical gelation temperature. The delivery device second state corresponds to a temperature range at and above the critical gelation temperature. The second temperature range is thus different, specifically higher, than the first temperature range. In the second state, the delivery device may mold or attain the shape of a location it is applied to, for example a body cavity, wound, etc.

[0077] The thermally reversible delivery device may include hyaluronic acid and at least one poloxamer with thermally reversible properties. The hydrogel may also include additional poloxamers that may alter the temperature at which the liquid to solid phase transition occurs.

[0078] A non-limiting example formula of a thermally reversible delivery device may be a hydrogel. An example hydrogel may include ZnCl2, NaCIO2, ascorbic acid, succinic acid, sodium chloride, poloxamer(s), and HA. Additional hydrogels are listed in the Examples.

[0079] An alternative component or delivery device of the treatment system may include a dressing. The formula may be applied via a dressing impregnated with the formula. The dressing may be a gauze dressing. The gauze may be applied wet and then may dry similar to a cast. The gauze dressing may be compressive. The formula present in the gauze may be slowly delivered to wound sites, including skin tears, dermal ulcers, venous stasis, arterial or neuropathic ulcers, or minor bums to improve healing and prevent new or worsening infections. The formula may be delivered to the gauze dressing by combining components of the formula (such as a non-limiting example of ZnCl2, NaClO2, ascorbic acid, starch, HA, and glycerin) to form a hydrogel with a consistency similar to a neomycin / polymyxin B / bacitracin ointment. The hydrogel may then be placed in plastic wrap or a plastic bag along with the gauze dressing and sealed so that the gauze may be impregnated with the hydrogel.

[0080] A non-limiting example of a dressing is shown in Fig. 2. The dressing 40 includes the formula described herein. The dressing may also include the thermos-reversible or temperature-dependent solution, as was described above.

[0081] Another type of delivery device may be a colloidal gel. A colloidal gel is a transient but arrested network of colloidal particles. The transition between liquid-like colloidal suspensions and the formation of colloidal gels depends on the volume fraction occupied by particles and the range and type of interaction between the particles.

[0082] Another type of delivery device may be a disposable cloth also known as wipes or wet wipes. Wet wipes typically include a cloth made from a non-woven fabric saturated with a solution. In this case, the cloth may be saturated with the charged formula disclosed herein. The charged formula may be included as an aqueous solution within the wipes or a hydrogel within the wipes.

[0083] The fabric of the wipes may be a natural or synthetic textile. Non-limiting example fabrics may include fibers of polyester, polypropylene, polyethylene, cotton, rayon, bamboo, the like, or a combination thereof. The fabric may be a non-woven fabric made directly from short fibers, long fibers, or both assembled in individual sheets. The boding of the fibers together may be facilitated chemically, mechanically, via heat, via solvent, or a combination thereof.

[0084] The fabric may also include one or more of the following components: softeners, lotions, perfumes, surfactants such as amphoteric surfactants, preservatives such as methylisothiazolinone, gluconolactone, the like, or their combination, pH buffers. The wet wipes may be flushable, sewer-safe, septic-safe, or a combination thereof. The wet wipes may be biodegradable or compostable.

[0085] The treatment system may include an intravenous delivery device to deliver the formula being an intravenous treatment solution. For example, the intravenous treatment may include zinc and sodium chlorite, C-peptide, Vitamin C (and other Vitamins), and other substances described herein.

[0086] The treatment system may include an injection such as an intra-articular injection or an injection entering a joint. The formula or a hydrogel containing the solution may be administered to a native joint or to a joint containing hardware, including but not limited to a total shoulder, knee, or hip replacement prosthesis, or hardware used to repair fractures.

[0087] The treatment system may include a catheter (e.g., an indwelling catheter or a catheter solely for irrigation). The catheter may be a catheter to the bladder and / or urethra to treat and / or prevent urinary tract infections or bacterial colonization.

[0088] The formula may be delivered through aerosolization for treatment of pulmonary infections such as pneumonia, particularly nosocomial or hospital-acquired pneumonia, bronchitis, or other respiratory infections. The device for distributing the formula may include, but is not limited to, a nebulizer, atomizer, mister, or other aerosol dispensing device.

[0089] The formula may be applied via a mechanism appropriate for the type of biomaterial present at the infected site. Non-liming examples of suitable dispensing, application or delivery mechanisms are disclosed in U.S. Patent Application 15 / 686,612, which is herein incorporated in its entirety. The formula may be applied to an infection site on the skin, for example, at a wound that is not healing due to a bacterial infection. In this example, the treatment system may include a pump such that the formula may be applied by pumping an effective amount onto the infected area or onto a bandage to be placed on the infected area.

[0090] As described above, the treatment system may be applied to infection sites including but not limited to surgical sites, body cavities, abscesses, acute or chronically occurring wounds, bum wounds, graft sites (e.g., skin grafts), joints, percutaneous pin sites, or any other biological tissue that is infected with microorganisms. The treatment system may be used to treat infections of any kind, and to manage, control, minimize, treat, and / or remove microorganisms, gram positive and gram negative bacteria, aerobic and anaerobic bacteria, viruses, fungi, or any other organisms / microorganisms having infectious properties.

[0091] The treatment system may also be applied onto body sites prone to odor generation due to presence of bacteria or fungi. Non-limiting example odor generation sites may include areas in proximity of sweat glands in the dermis such as armpits, inner thigs, small of the back, palmsof the hands, soles of the feet, forehead, etc. The secretion from the sweat glands typically includes a fluid, sweat, which contains proteins, lipids, and steroids. While sweat is naturally odorless, the skin microbiota metabolize the fluid which results in a production of malodorous byproducts causing unwanted body odor. Example of the skin microbiota producing body odor include aerobes, anaerobes, and fungi, such as Corynebacterium, Staphylococcus, Cutibacterium genera, Candida species, Aspergillus species, and Malassezia species. The treatment system may suppress, reduce, or minimize bacterial presence on the sites and thus reduce odor generation. The odor generation minimization or suppression by the treatment system may also apply to the infectious sites such as wounds.

[0092] Non-limiting examples of bacterial species that may be treated using the formula disclosed herein include ESKAPE pathogens, including Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter species. ESKAPE pathogens are nosocomial pathogens that are multidrug-resistant and virulent. They are associated with a high risk of mortality and impose a significant burden on the healthcare system worldwide.

[0093] Non-limiting examples of infections that are suitable for topical treatment with the formula include oral candidiasis and vaginal yeast infections caused by yeast such as Candida albicans', dandruff caused by Malassezia yeasts; Ringworm or Tinea caused by fungi including Trichophyton, Microsporum, or Epidermophyton', Intertrigo caused by Streptococcus or Corynebacterium minutissimum, fungi such as Candida albicans, or viruses; sores (cold and anogenital) and herpetic whitlow caused by the Herpes simplex virus; and rashes caused by the Varicella zoster virus. The topical treatment is also suitable for the above-mentioned skin microbiota including aerobic and anaerobic bacteria and fungi, such as Corynebacterium, Staphylococcus, Cutibacterium genera, Malassezia, Aspergillus, and Candida species.

[0094] As noted above, the formula may be used to treat surgical sites, wounds, and / or infections as well as odor generating sites in or on human bodies. The formula may also be used at infection sites as well as odor generating sites on or in animal bodies in the veterinary setting, as animals are subject to similar infection risks from open wounds, surgeries, and exposure to microorganisms .

[0095] EXAMPLES

[0096] Example 1: Prophylactic formulation

[0097] A non-limiting example of the herein-disclosed formulation for treating or preventing infections caused by planktonic bacteria was prepared as Example 1. The formula of Example 1 is in Table 1 below.

[0098] Table 1 - Example 1 formula

[0099] The zinc chloride and sodium chlorite may be present in equal molar amounts within the formula, or the zinc chloride and sodium chlorite may be present in non-equal molar amounts within the formula. The succinate / succinic acid serves as a buffer. Other buffers may be used.

[0100] Example 2: Treatment formulation

[0101] A non-limiting example of a formulation for treating infections caused by biofilm bacteria was prepared as Example 2. The formula of Example 2 is in Table 2 below.

[0102] Table 2 - Example 2 formula

[0103] The zinc chloride and sodium chlorite may be present in equal molar amounts, or the zinc chloride and sodium chlorite may be present in non-equal molar amounts within the formula of Example 2. The succinate / succinic acid serves as a buffer.

[0104] Example 3: Testing the effect of pH and ascorbic acid concentrations on effectiveness of the formula

[0105] Biofilm forming ESKAPE pathogens were treated with the formula containing 50 mM ZnCl2, with a pH of 4, 5 or 6, and 50 mM NaCIO2, as well as varying concentrations of ascorbic acid. In Tables 3-16 below, T5, T15, and T30 indicate data obtained after 5, 15, or 30 minutes of treatment respectively.

[0106] It was observed that a pH of 4 and an ascorbic acid concentration of 1 or 3 mM were most effective for eradicating biofilm bacteria at 5 minutes after treatment.

[0107] Planktonic ESKAPE pathogens were treated with another embodiment of the formula containing 25 mM ZnCl2, with a pH of 4, 5 or 6, and 25 mM NaCIO2, as well as varying concentrations of ascorbic acid. The formula was effective at eradicating planktonic bacteria at 1 minute after treatment when the concentration of ascorbic acid in the formula was 0 or 3 mM.

[0108] In the data sets below, bold font indicates a log biofilm reduction (kill) value of less than 1. Underlined_with _ dashed line indicates a log value between 1 and 2. Underlined with solid line indicates a log biofilm reduction (kill) value between 2 and 3. Italics indicates a value higher than 3. “CE” denotes “Complete Eradication.” “AA00” - “AA5” indicates concentration of ascorbic acid (AA) in [mM] in the formulas.

[0109] Tables 3-5 - Data for Acinetobacter baumannii

[0110] Tables 6-9 - Data for Pseudomonas aeruginosa

[0112] Tables 12-14 - Data for Klebsiella pneumoniae

[0113] Tables 15, 16 - Data for Enterococcus faecium

[0114] Treatment of additional planktonic bacterial species with the formula containing 25 mM ZnCl2, pH 4, and 25 mM NaCIO2, as well as varying concentrations of ascorbic acid was prepared. This formula was efficient at eradicating planktonic bacteria at 1 minute after treatment when the concentration of ascorbic acid in the formula was 0 or 1.5 mM. The results are shown in Table 17 below.

[0115] Table 17 - Data for Escherichia coli

[0116] Table 18 - Data for Staphylococcus epidermidis[0117[ Example 4: Testing effectiveness of the formula depending on the length of shelf storage time

[0118] The effect of shelf storage time (after combining ZnCl2with NaCIO2) on the effectiveness of various embodiments of the formula was tested using ESKAPE pathogen bacteria. The formulas included 50 mM ZnCl2with either 0, 0.5, 1.5, or 5 mM ascorbic acid and 50 mM NaCIO2. The indicated bacteria were exposed to the formula for 5 or 15 minutes, at which time the number of bacterial colonies killed was measured. In the tables below, “CE” denotes complete eradication, “AA” denotes concentration of ascorbic acid, “C” denotes concentration of NaCIO2, and “Z” denotes concentration of ZnCl2.

[0119] Table 19 - Data for Enterococcus faecium

[0120] Table 20 - Data for Staphylococcus aureus

[0121] Table 21 - Data for Klebsiella pneumoniae

[0122] Table 22 - Data for Acinetobacter baumannii

[0123] Table 23 - Data for Pseudomonas aeruginosa

[0124] Example 5: Testing formula stored for 7 weeks after the components were combined

[0125] Formulations with differing concentrations of the various components were tested for their efficiency in eradicating ESKAPE pathogen bacteria 7 weeks after the components were combined to produce the formulation. The rows refer to the concentration of ZnCl2in the formulation (rnM) designated as “Z,” and the columns refer to the concentration of NaClO2in theformulation (mM), designated as “C.” The bacteria were exposed to each formulation for 1, 5, 15, or 30 minutes (T1, T5, T15, T30), at which time the number of bacteria killed was measured.

[0126] Tables 24, 25 - Data for Pseudomonas aeruginosa

[0127] Tables 26, 27 - Data for Staphylococcus aureus

[0128] Example 6: Testing stability and efficacy of solution with prolonged storage after combining

[0129] Regarding formulas listed in Table 28, all of the formulas listed in Rows 1-5 were tested on the same day. The formula in Row 1 was made and left on the shelf for 4 months before testing. The NaClO2and ZnCl2used to make the formula in Row 2 were made and left on the shelf for 4 months, and combined on the day of testing. The NaCIO2used to make the formula in Row3 was made and left on the shelf for 4 months. The ZnCl2used to make the formula in Row 3 was made and combined with the four-month-old NaCIO2on the day of testing. The ZnCl2used to make the formula in Row 4 was made and left on the shelf for 4 months. The NaCIO2used to make the formula in Row 4 was made and combined with the four-month-old ZnCl2on the day of testing. The solutions used to make the formula in Row 5 were made and combined on the day of testing.

[0130] Table 28 — Data for Pseudomonas aeruginosa biofilms

[0131] Example 7: Preparation of a non-limiting example of a PVA / HA hydrogel

[0132] Preparation of 20% polyvinyl alcohol (PVA): 80.01 g (80 g) of PVA was added to320 mL of distilled water on a hot plate magnetic stirrer at about 80-90°C. The solution was stirred until fully dissolved, then allowed to cool to room temperature. Periodic manual stirring was used to prevent the surface from forming a thin membrane. Any evaporated water was re-added and mixed thoroughly once the solution was cooled.

[0133] Preparation of 2% Hyaluronic Acid (HA) Solution: 2.5000 g (2.5 g) of hyaluronic acid was added to about 122.5 mL of distilled water. The solution was stirred until fully hydrated.

[0134] Preparation of Hydrogel Solution: 20% PVA and 2% HA were prepared as above, succinic acid, ascorbic acid, sodium chloride, and zinc chloride were weighed out as per Table 29 below and dissolved in a small amount of distilled water. The mixture was added to the 2% HA solution and thoroughly mixed. The sodium chlorite was weighed and dissolved in a small amount of distilled water and added to the 2% HA solution as well. The solution was quickly diluted to 500 mL with the 20% PVA solution. The solution was mixed thoroughly, and the pH was adjusted to 5.34 @ 33.3°C using 10% NaOH and 10% HC1. The solution was stirred until the pH was stable. The final concentration of this solution was 15% polyvinyl alcohol, 0.5% hyaluronic acid, 7.5 mMsuccinic acid, 30 mM sodium chloride, 6 mM ascorbic acid, 200 mM zinc chloride, and 160 mM sodium chlorite. The final concentration of sodium chlorite was 160 mM instead of 200 mM because the highest available purity of sodium chlorite is 80%.

[0135] Preparation of Hydrogel: A gasket design was created using MatterControl Software. The gaskets were 3D-printed on a Dremel 3D45 3D printer, and silicone molds were made from the printed parts. The Hydrogel Solution was poured into the silicone molds and put through 3 freeze-thaw cycles, which consisted of placing the samples into the -20°C freezer for 18 hours, thawing the samples at 5 °C for 6 hours, then repeating the process two more times. This caused the PVA molecules to cross-link, forming a solid hydrogel.

[0136] Table 29 - Hydrogel formula

[0137] Example 8: Preparation of a non-limiting example of a starch HA hydrogel

[0138] Succinic acid, ascorbic acid, sodium chloride, and zinc chloride were added to 87 mL of distilled water. The pH was adjusted to about 5.0 with 10% NaOH. Sodium chlorite was added to the solution, and the pH was adjusted to 5.21 with sodium hydroxide and HA (1% and 10%). 8.0013 (8 g) of starch was added to the solution and pH was rechecked. The pH remained at 5.21 , so the solution was heated on a hot plate magnetic stirrer until the starch gelatinized (about 60-80°C). The gel was removed from the heat, and 2.0032 g of HA was added and mixed thoroughly with a glass stir rod. The mixture was placed in the fridge (5°C) to help it cool. Mixingwas continued until the HA was fully hydrated into the gel. The gel had a consistency of an over- the-counter triple antibiotic ointment.

[0139] Table 30 - Hydrogel formulation

[0140] Example 9: Preparation of starch, HA, glycerin hydrogel impregnated gauze

[0141] Succinic acid, ascorbic acid, sodium chloride, and zinc chloride was added to about 75 mL of distilled water. pH was adjusted to about 5.0 with 10% NaOH. Sodium chlorite was added to the solution, and the pH was adjusted to 5.02 with sodium hydroxide and hydrochloric acid (1% and 10%). 8.0021 (8 g) of starch was added to the solution and pH was rechecked. The pH remained at 5.02, so the solution was heated on a hot plate magnetic stirrer until the starch gelatinized (~60-80°C). The gel was removed from the heat and 2.0005 g of HA was added and mixed thoroughly with a glass stir rod. About 5 mL of glycerin was also stirred into the mix. The mixture was placed in the freezer (-20°C) to help it cool. Mixing was continued until the HA was fully hydrated into the gel. The gel had a consistency of an over-the-counter triple antibiotic ointment. The hydrogel was placed in a container with gauze to form a hydrogel-impregnated gauze.

[0142] Table 31 - Hydrogel formula

[0143] Example 10: Preparation of Poloxamer 407 and Poloxamer 188 mixes for thermal reversible hydrogels

[0144] Solutions were prepared by weighing out (in order) HA, Poloxamer 407, and Poloxamer 188 into separate 50 mL conicals. The powder was mixed by twisting the conical. Subsequently, into separate 50 mL beakers, the succinic acid, ascorbic acid, sodium chloride, and zinc chloride were mixed with approximately 25 mL of distilled water. 8 drops of 10% sodium hydroxide was added, then the sodium chlorite was added, and the volume was adjusted to about 23 mL and about 28 mL, respectively, with distilled water. The pH and temperature were recorded. The solutions were then added to the 50 mL conicals containing the poloxamers and HA. Using a glass stir rod, the solutions were stirred to break up the poloxamer / HA powders. The volume was then brought up to the 50 mL mark using distilled water. The conicals were closed and placed in the refrigerator to finish dissolving overnight. These solutions, once fully dissolved / hydrated, were thermally reversible at temperatures of around 21-23 °C.

[0145] Table 32 - Thermally reversible hydrogel formula

[0146] Table 33 - Thermally reversible hydrogel formula

[0147] Examples 11-18 - Hydrogels

[0148] Examples 11-18 represent hydrogels prepared according to the methods described above. Formulas and visual development of bacterial inhibition is depicted in Figs. 3-10. In Examples 11-18, the hydrogels were placed onto a lawn of Staphylococcus aureus bacteria and imaged after 1, 2, 3, 4, or 5 days. The inner and outer radii and the weight were measured. The inner radii represent the zones of complete inhibition, while the outer radii represent zones of partial inhibition (from the hydrogel to where minimal bacteria transitions to a complete lawn of bacteria). The weight of the hydrogel may decrease as the chlorite / zinc chloride elutes out. The weight may increase as material from the agar or bacteria on the agar enters the hydrogel.

[0149] Example 11 included hydrogels made with a formulation of the formula containing 50 mM ZnCl2, 50 mM NaCIO2, 1.5 mM ascorbic acid, and 10% PVA. Fig. 3 shows test results for Example 11 at 24-120 hours or 5 days of bacterial inhibition. Example 12 included hydrogels made with a formulation of the formula containing 50 mM ZnCl2, 50 mM NaCIO2, 1.5 mM ascorbic acid, and 10% PVA. Fig. 4 shows Example 12 imaged after 1-5 days. Example 13 included hydrogels made with a formulation of the formula containing 50 mM ZnCl2, 50 mM NaCIO2, 1.5 mM ascorbic acid, 7.5% PVA, and 0.5% HA. Fig. 5 shows results after 24-96 hours. Example 14 included hydrogels made with a formulation of the formula containing 100 mM ZnCl2, 100 mM NaCIO2, 3 mM ascorbic acid, and 10% PVA. Fig. 6 shows results after 1-5 days. Example 15 included hydrogels made with a formulation of the formula containing 25 mM ZnCl2, 25 mM NaCIO2, 0.75 mM ascorbic acid, and 10% PVA. Fig. 7 shows results after 1, 2 and 3 days. Example 16 included hydrogels made with a formulation of the formula containing 50 mM ZnCl2, 50 mM NaCIO2, 1.5 mM ascorbic acid, 5% PVA, and 1% hyaluronic acid (HA). Fig. 8 shows results of bacterial inhibition after 1 , 2, and 3 days. Example 17 included hydrogels made with a formulation of the formula containing 50 mM ZnCl2, 50 mM NaCIO2, 1.5 mM ascorbic acid, 2.5% PVA, and 1.5% hyaluronic acid (HA). Fig. 9 shows results after 1-3 days. Example 18 included hydrogels made with a formulation of the formula containing 100 mM ZnCl2, 100 mM NaCIO2, 3 mM ascorbic acid, and 2% hyaluronic acid (HA). Example 18 was PVA free. Fig. 10 shows imaged results after 1 day.

[0150] Example 19: Preparation of 2% Hyaluronic Acid Hydrogel with Cl 00 Z100 A3 Solution

[0151] Using an Analytical Balance, the following were weighed out to give the desired concentrations listed in the tables below. Preparation of ~2% hyaluronic acid: 1.0002 g (1 g) of hyaluronic acid was added to about 45 mL of distilled water and mixed until fully hydrated into a thick gel-like viscous solution.

[0152] Preparation of Hydrogel: 0.0464 g (0.0443 g) succinic acid, 0.0867 g (0.0877 g) sodium chloride, 0.0252 g (0.0264 g) ascorbic acid, and 0.6843 g (0.6815 g) zinc chloride was added to the ~2% hyaluronic acid solution. The pH was adjusted to around 4.5 using 1% and 10% sodium hydroxide and hydrochloric acid. 0.4540 g (0.4522 g) sodium chlorite was then added anddissolved into the solution. The pH was adjusted to 4.92 at 21.2°C (pH ~5) using 1% and 10% sodium hydroxide and hydrochloric acid. Distilled water was added to make up the volume to 50 mL. The solution was thoroughly mixed into a viscous, light green, hydrogel; with the consistency of a gel shampoo. The final concentration of the solution was 2% hyaluronic acid, 7.5 mM succinic acid, 30 mM sodium chloride, 3 mM ascorbic acid, 100 mM zinc chloride, and 80 mM sodium chlorite. NOTE: The final concentration of sodium chlorite is 80 mM instead of 100 mM because the highest available purity of sodium chlorite was 80%.

[0153] Table 34 - Hyaluronic acid hydrogel formula

[0154] Example 20: Optimized preparation of PVA HA hydrogels

[0155] Example 20 provides data and procedural details involving the preparation of PVA HA hydrogels. Preparation of 20% PVA: 20.02 g (20 g) of PVA was added to 80 mL of DMSO and stirred with heat on a hotplate. The solution was stirred for approximately 15 minutes, however, no changes were being observed with the PVA. Preparation of 2% hyaluronic acid solution: 0.5051 g (0.5 g) of hyaluronic acid was added to about 24.5 mL of distilled water. The solution was stirred until fully hydrated. Preparation of the main solution: 20% PVA and 2% HA were prepared as above. Since no changes were observed in the PVA, the entire solution was added to the 2% HA solution. The solution stirred on the hotplate in the fume hood for an additional hour. The solution began to thicken, so more water was added back up to a volume of about 120mL.

[0156] The succinic acid, ascorbic acid, NaCl, and ZnCl2were weighed out and dissolved in the hot mixture. 16 drops of 10% NaOH was added to the solution. Periodic manual stirringwith a glass stir rod was done to prevent a film from forming on the surface. The solution was allowed to stir and cool back down to room temperature. The NaC102was weighed and dissolved in the mixture once cool. The solution was mixed thoroughly and the pH was adjusted to 5.04 @ 35.1 °C using 10% NaOH and 10% HC1. The solution was stirred until pH was stable. The solution was placed in a vacuum chamber to attempt to remove the bubbles. The solution was poured into molds and the freeze-thaw cycles were started.

[0157] Table 35 - Freeze-thaw cycle data for PVA HA hydrogels of Example 20 samples

[0158] After thawing, it was observed that the samples may not have fully hydrated the PVA before forming the gels. The gels were used for zone of inhibition testing. The samples had a silicone-like feel. Very strong and slightly transparent. After sitting out for 1 day, 6 samples were vacuum sealed to check long term storage and malformation. Removed from freezer on day 6. Sealed in vacuum sealed bags on day 7 at 8:15am - 8:40am.

[0159] Table 36 - PVA HA hydrogel formula

[0160] Preparation of 20% PVA in 80:20 DMSO:water: 20.00 g (20 g) of PVA was added to 80 mL of 80:20 DMSO:water and stirred with heat on a hotplate. The solution was stirred for about 2 hours at 80-100°C. The solution became clear and was allowed to cool while stirring for about 1 hour. The solution was reheated back to a syrup-like consistency and poured into molds (plates) while still hot. The solution was then left to cool in the hood before covering, sealing, and placing in the -20°C freezer to begin the freeze-thaw process.

[0161] Table 37 - Freeze-thaw cycle data for PVA HA hydrogels

[0162] Preparation of 20% PVA: 20.05 g (20 g) of PVA was added to 80 mL of an 80:20 DMSO:water mixture and stirred with heat on a hotplate. The solution was stirred until fully hydrated. Preparation of 2% hyaluronic acid solution: 0.5006 g (0.5 g) of hyaluronic acid was added to about 24.5 mL of an 80:20 DMSO: water mixture. The solution was stirred until fully hydrated.

[0163] Preparation of Hydrogel: 20% PVA and 2% HA were prepared as above. The ZnCl2, NaCl, ascorbic acid, and succinic acid were weighed out and dissolved in the 2%HA solution and thoroughly mixed. The pH was raised with a few drops of 10% NaOH. The 20%PVA solution was added to the 2% hyaluronic acid solution and thoroughly mixed to a final volume of 100mL. The NaClO2was weighed and added mixed solution. The sodium chlorite spontaneously combusted from the heat of the solution. The solution was left in the fume hood until the reaction was no longer occurring.

[0164] Table 38 - PVA HA hydrogel formula

[0165] Preparation of 20% PVA: 20.00 g (20 g) of PVA was added to 80 mL of DMSO and stirred with heat on a hotplate. The solution was stirred until fully hydrated. Preparation of 2% hyaluronic acid solution: 1.0001 g (1.0 g) of hyaluronic acid was added to about 49 mL of distilled water. The solution was stirred until fully hydrated.

[0166] Preparation of hydrogel: The above were added to 25mL of the 2% Hyaluronic Acid solution in the following order: succinic acid, ascorbic acid, sodium chloride, and zinc chloride. Water was used to rinse each ingredient into solution. The pH was adjusted to about 4.5, then the sodium chlorite was added and mixed well. Final volume was about 35 mL. Once cooled to about 70°C, the 20% PVA solution in DMSO was added to make a final volume of about 115 mL. The solution was mixed well and poured into molds to begin the freeze-thaw cycles.

[0167] Table 39 - Freeze-thaw cycle data for PVA HA hydrogels

[0168] Table 40 - PVA HA hydrogel formula

[0169] Preparation of 20% PVA: 20.06g (20g) of PVA was added to 80mL of an 80:20 DMSO: Water mixture and stirred with heat on a hotplate. Solution was stirred until fully hydrated. (NOTE: Next time dissolve in 80mL DMSO Only). Preparation of 2% Hyaluronic Acid Solution: 0.5008g (0.5g) of hyaluronic acid was added to ~24.5mL of distilled water. The solution was stirred until fully hydrated. Preparation of main solution: Succinic acid, ascorbic acid, sodium chloride, and zinc chloride was dissolved into the 2% hyaluronic acid solution. pH was adjusted to -5.00 using 1% and 10% sodium chloride and hydrochloric acid. Sodium Chlorite was then added, and the pH was adjusted from 5.48 to 4.96. Once cooled to ~55°C, the 20% PVA solution was mixed into the 2% The solution was mixed thoroughly, then poured into molds to form the hydrogel shells. The mold was placed in the freezer to begin the freeze-thaw cycles. NOTE: The solution needed to remain around 50-60°C to prevent cross-linking.

[0170] Table 41 - Freeze-thaw cycle data for PVA HA hydrogels

[0171] Table 42 - PVA HA hydrogel formula

[0172] Preparation of 2% hyaluronic acid solution: 0.4997 g (0.5 g) of hyaluronic acid was added to ~24.5 mL of distilled water. The solution was stirred until fully hydrated. Preparation of 20% PVA: 20.02 g (20 g) of PVA was added to 80 mL of 80:20 DMSO:water and stirred with heat on a hotplate. The solution was stirred until fully dissolved. Preparation of hydrogel: succinic acid, ascorbic acid, sodium chloride, and zinc chloride were added to the 2% HA solution. The pH was raised to -4.9 using 10% sodium hydroxide. Sodium chlorite was then added to the solution and mixed thoroughly. pH was adjusted to 5.00 @ 27.3°C with 10% hydrochloric acid. While still warm, but not hot, the 20% PVA solution was slowly added to the 2%HA mixture to a volume of 100 mL. The solution was stirred thoroughly and kept on the hotplate to keep warm. The solutionwas injected with a syringe into small mold locations and the rest was poured into molds and placed in the -20°C freezer to begin freeze-thaw cycles.

[0173] Table 43 - Freeze-thaw cycle data for PVA HA hydrogels

[0174] Table 44: PVA HA hydrogel formula

[0175] Example 21: Optimizing preparation of alginate hydrogels:

[0176] Example 21 provides data and procedural details involving the preparation of alginate hydrogels. Sodium alginate hydrogel: Day 1: Preparation of ~1% Calcium Carbonate: 0.4990 g (0.5 g) calcium carbonate was added to 49 mL of distilled water. Concentrated hydrochloric acid was added dropwise to the solution until fully dissolved. pH was adjusted to ~3.6 using 10% Sodium Hydroxide and 10% Hydrochloric Acid. Preparation of 1% Sodium Alginate: 1.0036 g (1 g) sodium alginate was added to 99 mL of distilled water and stirred until dissolved. Preparation of test blank gel: A portion of the ~1% calcium carbonate solution was poured into a bottle. The 1% sodium alginate was drawn into a syringe and droplets were droppedinto the calcium carbonate solution. The droplets formed bead-like gels almost instantly. The sodium alginate solution was then injected as a stream into the calcium carbonate solution. The stream turned into a noodle-like gel. The gels were left out in open air to dry out.

[0177] Day 5 - The gels that were left out were completely dried to a film. A few were left under a vial. These gels remained gels, but shrank in size, meaning that the drying was slowed under the vial. Some distilled water was added to the dried films.

[0178] Day 6 - The dried films appeared to have rehydrated a little. More distilled water was added to submerge the jelly-like film. Calcium carbonate solution was poured into molds. Sodium alginate was added to the mold to see if it would form to the mold. It did not. The same was tried by pouring sodium alginate into molds and then placed calcium carbonate into the mold. Gel formed, however, it was not in the shape of the mold. Also, half of a gauze was soaked in calcium carbonate, and the other half was soaked in sodium alginate. The opposites were added to each half to trigger crosslinking. The half that was first soaked in calcium carbonate formed a globby gel on one side of the gauze. The half that was soaked in sodium alginate first formed a gel within the gauze that appeared to be evenly distributed among the gauze. The gauze was rinsed with distilled water and placed in a petri dish to dry out.

[0179] Day 7 - The jelly-like film appeared to have fully rehydrated back to the original flexible gel. In addition, the gauze samples were completely dried out. Distilled water was sprayed over them and the gauze immediately became flexible again. It appeared that the sodium alginate film began to rehydrate.

[0180] Day 25 - 1% sodium alginate made on Day 1 was added into a gasket mold. A few drops of 1% calcium carbonate were added to the sodium alginate. About 2 minutes were allowed to pass to allow gelation to occur. A gasket shaped hydrogel formed with rough edges. 1% sodium alginate was applied to some gauze in the shape of a pad. 1% calcium carbonate was dripped over the alginate, allowing gelation to occur. A gel pad was successfully impregnated into the gauze.

[0181] Day 26 - sodium alginate pad dried out overnight, making a stiff absorbant pad.

[0182] Sodium alginate hydrogel: Day 1: preparation of ~1% calcium carbonate: 1.0058 g (1.0 g) calcium carbonate was added to 99 mL of distilled water. Concentrated hydrochloric acid was added dropwise to the solution until fully dissolved. Zinc chloride was added to the solution, and pH was adjusted to about 3.3 using 10% sodium hydroxide and 10% hydrochloric acid. Preparation of 2% sodium alginate: 1.0013 g (1 g) sodium alginate was added to 49 mL of distilled water and stirred until dissolved. Preparation of 1% sodium alginate: 0.5031 g (0.5 g) sodium alginate was added to 49.5 mL of distilled water and stirred until dissolved. Preparation of 1% hyaluronic acid / 1% sodium alginate: 0.5026 g (0.5 g) hyaluronic acid was hydrated in 49 mL of distilled water. 0.5029 g (0.5 g) sodium alginate was added and stirred until hydrated.

[0183] Table 45: alginate hydrogel formula

[0184] The outlined solution above was formed into 5 4x4in pads on gauze, then vacuum sealed individually and stored protected from light.

[0185] Day 1 : preparation of 1% hyaluronic acid / 1% sodium alginate: 0.5005 g (0.5 g) hyaluronic acid was hydrated in 49 mL of distilled water. 0.5006 g (0.5 g) sodium alginate was added and stirred until hydrated. pH was measured to be 7.03 @ 24.6°C.

[0186] Table 46: alginate hydrogel formula

[0187] Sodium alginate hydrogel: Day 1 : preparation of 1% sodium alginate: 0.5015 g (0.5 g) sodium alginate was added and stirred until hydrated. Preparation of 1% calcium chloride, 50 mM zinc chloride solution: The below zinc chloride and caclium chloride amounts were added to 50 mL of distilled water. pH was adjusted to 3.40 at 21.6°C with 10% and 1% NaOH and HC1. Once all of the sodium alginate was hydrated, succinic acid, ascorbic acid, and sodium chloride was added to the solution. pH was checked and adjusted to ~4.6. The sodium chlorite was added to the solution and the pH changed to -5.62 @ 26.0°C. The solution was then impregnated into a gauze sheets and the 1% calcium chloride, 50 mM zinc chloride solution was dropped onto the gel solution with a dropper to cause gelation into the gauze. Sheets were bagged and testing on wounds were begun at 3:45 pm.

[0188] Table 47 - alginate hydrogel formula

[0189] Sodium alginate hydrogel: Day 1 : preparation of 1 % sodium alginate: 0.5030 g (0.5 g) sodium alginate was added and stirred until hydrated. Preparation of 1% calcium chloride, 50 mM zinc chloride solution: The below zinc chloride and calcium chloride amounts were added to 100 mL of distilled water. pH was adjusted to 3.40 at 23.0°C with 10% and 1% NaOH and HC1. Solution was transferred into conical flasks until needed. Once all of the sodium alginate was hydrated, succinic acid and sodium chloride was added to the solution. pH was determined to be 4.25 at 26.0°C. The solution was transferred to a conical flask and stored without sodium chlorite overnight. Sodium chlorite was added to the solution and the pH was measured to be 4.42 @ 22.2°C. Solution was left at that pH and was transferred to a syringe. Solution was impregnated into small, square pieces of gauze and gelled by using the 1% calcium chloride, 50 mM zinc chloride solution.

[0190] Table 48 - alginate hydrogel formula

[0191] Sodium alginate hydrogel: Day 1 : preparation of 1% sodium alginate: 0.5042 g (0.5 g) sodium alginate was added and stirred until hydrated. Preparation of 1% calcium chloride, 50 mM zinc chloride solution: used solution prepared 14 days earlier. The solution was transferred into conical flasks until needed. Once all of the sodium alginate was hydrated, succinic acid and sodium chloride was added to the solution. The solution was stirred overnight to fully hydrate and mix. Sodium chlorite was added to the solution and the pH was measured to be 4.64 @ 25.6°C. The solution was left at that pH and was transferred to a syringe. The solution was impregnated into small, square pieces of gauze and gelled by using the 1% calcium chloride, 50 mM zincchloride solution. Remaining 1% CaCl2, 50 mM ZnCl2was dumped out and gels were rinsed with distilled water. VI SA-2 gels were stored for use in zone of inhibition testing.

[0192] Day 8 - Gels were made by weighing 1g into wells containing gauze. 1% calcium chloride 50 mM zinc chloride solution was added to each well to gel the solutions. The gels were rinsed with distilled water.

[0193] Table 49 - alginate hydrogel formula

[0194] Sodium alginate hydrogel: Day 1: preparation of 2% sodium alginate: 2.0002 g (2 g) sodium alginate was added and stirred until hydrated. Preparation of 3% calcium chloride, 50 mM zinc chloride solution: the below zinc chloride and calcium chloride amounts were added to 100 mL of distilled water. pH was adjusted to 3.39 at 24.2°C with 10% and 1% NaOH and HC1. The solution was transferred into conical flasks until needed.

[0195] Day 2: Once all of the sodium alginate was hydrated, succinic acid and sodium chloride was added to the solution. Sodium chlorite was added to the solution and the pH was measured to be 5.05 @ 24.3°C. The solution was left at that pH and was transferred to a syringe. V2SA-1 gels were made by weighing 1 g into wells containing gauze. 3% calcium chloride 50 mM zinc chloride solution was added to each well to gel the solutions. The gels were rinsed with distilled water.

[0196] Table 50 - alginate hydrogel formula

[0197] Sodium alginate hydrogel: Day 1: preparation of 2% sodium alginate: 5.0180 g (5 g) sodium alginate was added to 245 mL distilled water and stirred until hydrated. Preparation of 3% calcium chloride, 50 mM zinc chloride solution: the below zinc chloride and calcium chloride amounts were added to about 240 mL of distilled water. pH was adjusted to 3.38 at 25.4°C with 10% and 1% NaOH and HC1. The solution was transferred into conical flasks. Once all of the sodium alginate was hydrated, succinic acid and sodium chloride was added to the solution. Sodium chlorite was added to 50 mL of the solution and the pH was measured to be 4.46 @ 25.0°C for the 12.5 mM solution. The solution was left at that pH and was transferred to a syringe. Sodium chlorite was added to 50 mL of the solution and the pH was measured to be 4.55 @ 24.6°C for the 25 mM solution. The solution was left at that pH and was transferred to a syringe. Sodium chlorite was added to 50 mL of the solution and the pH was measured to be 4.96 @ 24.3°C for the 75 mM solution. The solution was left at that pH and was transferred to a syringe. V2SA-1 gels were made by weighing 1 g into wells containing gauze. 3% calcium chloride 50 mM zinc chloride solution was added to each well to gel the solutions. The gels were rinsed with distilled water.

[0198] Table 51 - alginate hydrogel formula

[0199] Sodium alginate hydrogel: Day 1: preparation of 2% sodium alginate: 2.0014 g (2 g) sodium alginate was added to 98 mL distilled water and stirred until hydrated. Preparation of 3% calcium chloride, 50 mM zinc chloride solution: The below zinc chloride and calcium chloride amounts were added to about 96 mL of distilled water. pH was adjusted to 3.32 at 25.8°C with 10% and 1% NaOH and HC1. The solution was transferred into conical flasks until needed.

[0200] Table 52 - alginate hydrogel formula

[0201] Sodium alginate hydrogel: Day 1: preparation of 2% sodium alginate: 2.0032 g (2 g) sodium alginate was added to 98 mL distilled water and stirred until hydrated. Preparation of 3% calcium chloride, 50 mM zinc chloride solution: the below zinc chloride and calcium chloride amounts were added to about 96 mL of distilled water. pH was adjusted to 3.32 at 26.6°C with 10% and 1% NaOH and HC1. The solution was transferred into conical flasks. Once all of the sodium alginate was hydrated, succinic acid and sodium chloride were added to the solution.Sodium chlorite was added to 50 mL of the solution and the pH was measured to be 4.43 @ 26.1 °C for the 12.5 mM solution. The solution was left at that pH and was transferred to a syringe. Sodium chlorite was added to 50 mL of the solution and the pH was measured to be 4.58 @ 26.2°C for the 25 mM solution. The solution was left at that pH and was transferred to a syringe. V2SA-1 12.5 mM and 25 mM solutions were weighed out to about 1 g into molds. The 3% CaCl250 mM ZnCl2solution was added to the molds to gel the V2SA-1 and allowed to sit for about 5 minutes. The remaining solution was drained, and the gels were rinsed with distilled water. The gels were vacuum sealed and placed in the fridge until use.

[0202] Table 53- alginate hydrogel formula

[0203] Sodium alginate hydrogel: Day 1: preparation of 2% sodium alginate: 16.01 g (16 g) sodium alginate was added to 784 mL distilled water and stirred until hydrated. Preparation of 3% calcium chloride, 50 mM zinc chloride solution: the below zinc chloride and calcium chloride amounts were added to about 963 mL of distilled water. pH was adjusted to 3.37 at 27.9°C with 10% and 1% NaOH and HC1. The solution was transferred into conical flasks until needed.

[0204] Day 6: Once all of the sodium alginate was hydrated, succinic acid, and sodium chloride were added to the solution. Sodium chlorite was added to 250 mL of the solution and the pH was measured to be 4.52 @ 23.6°C for the 12.5 mM solution. The solution was left at that pH and was transferred to a syringe. Sodium chlorite was added to 250 mL of the solution and the pH was measured to be 4.58 @ 23.3 °C for the 25 mM solution. Solution was left at that pH and wastransferred to a syringe. V2SA-1 12.5 mM and 25 mM solutions were weighed out to about 1 g into molds. The 3% CaCl250 mM ZnCl2solution was added to the molds to gel the V2SA-1 and allowed to sit for about 5 minutes. The remaining solution was drained, and the gels were rinsed with distilled water. The gels were vacuum sealed and placed in the fridge until use. Any leftover solution was bottle / vialed and saved until future use.

[0205] Table 54 - alginate hydrogel formula

[0206] Example 22: Preparing poloxamer-alginate hydrogels:

[0207] Example 22 provides data and procedural details involving the preparation of poloxamer-alginate hydrogels.

[0208] Table 55 - poloxamer-alginate hydrogel formula

[0209] The above was prepared by first hydrating the hyaluronic acid in 96 mL of distilled water. Once fully hydrated, the sodium alginate was added to the solution and stirred until fully dissolved. This was labelled as 3% SA solution. To a separate beaker, 2.0008 g poloxamer 188 was added and hydrated in 94 mL distilled water. Then 3.0013 g calcium carbonate and 1.3657 g zinc chloride was thoroughly mixed in. Alternative mixtures were then explored. NOTES: Making the solution with too high of a poloxamer concentration weakened the gel.

[0210] Preparation of Poloxamer 407 solution: 15.24 g (15 g) of poloxamer 407 was added to 85 mL of distilled water and dissolved in the fridge / freezer. Preparation of Poloxamer 188 Solution: 5.07 g (5 g) of Poloxamer 188 was added to 95 mL of distilled water and dissolved in / out of the fridge. Preparation of Solution A: 50 mL of 5% Pl 88 solution was mixed with 50 mL of 3% SA solution. The above succinic acid and sodium chloride was added to the solution. pH was adjusted to ~4.7 and then sodium chlorite was added to the Pl 88 solution and fully dissolved before adding the SA. pH of this solution was determined to be 5.66 @22.3°C. Preparation of Solution B: 3.0068 (3 g) Calcium Carbonate and the above zinc chloride was added to the 15% Poloxamer 407. Preparation of Hydrogel: 5 mL of Solution A was mixed with 5 mL of Solution B and stirred thoroughly. The solution was then covered and placed in the incubator at 37°C for 2 hours to complete gelation. Upon initial mixing, it appeared that the solution still gelled right away. Into Incubator: 2:00pm. Out of Incubator: 4:00pm. Incubator Temperature: 37°C. Sample was removed from the incubator. The gel was weak and improperly shaped.

[0211] While exemplary embodiments are described above, it is not intended that these embodiments describe all possible forms encompassed by the claims. The words used in the specification are words of description rather than limitation, and it is understood that various changes can be made without departing from the spirit and scope of the disclosure. As previously described, the features of various embodiments can be combined to form further embodiments that may not be explicitly described or illustrated. While various embodiments could have been described as providing advantages or being preferred over other embodiments or prior artimplementations with respect to one or more desired characteristics, those of ordinary skill in the art recognize that one or more features or characteristics can be compromised to achieve desired overall system attributes, which depend on the specific application and implementation. These attributes can include, but are not limited to cost, strength, durability, life cycle cost, marketability, appearance, packaging, size, serviceability, weight, manufacturability, ease of assembly, etc. As such, to the extent any embodiments are described as less desirable than other embodiments or prior art implementations with respect to one or more characteristics, these embodiments are not outside the scope of the disclosure and can be desirable for particular applications.

Claims

WHAT IS CLAIMED IS:

1. A treatment system for a microbial infection comprising: a solution including: a source of ions in the amount of about 25-225 mM; and a source of an oxidizing agent in the amount of about 25-225 mM; water; and a delivery device comprising polysaccharide(s) forming a three-dimensional polymeric biphasic network housing the solution.

2. The system of claim 1, wherein the polysaccharide(s) include polyvinyl alcohol.

3. The system of claim 1, wherein the network forms a body with a centrally-located aperture.

4. The system of claim 3, wherein the body has a raised portion adjacent the centrally- located aperture.

5. The system of claim 3, wherein the body includes a second aperture running from an edge of the body to the centrally-located aperture.

6. The system of claim 1, wherein the source of ions includes zinc chloride (ZnCh).

7. The system of claim 1, wherein the oxidizing agent includes sodium chlorite (NaC102).

8. The system of claim 1, wherein the delivery device is a hydrogel with a uniform thickness.

9. A treatment system comprising: an aqueous solution including:a source of ions in the amount of about 25-225 mM; and a source of an oxidizing agent in the amount of about 25-225 mM; and a thermally reversible delivery device housing the solution, the delivery device having a first state of being a liquid at a first temperature range and a second state of being a gel at a second temperature range, the second temperature range being different than the first temperature range.

10. The system of claim 9, wherein the second temperature range is higher than the first temperature range.

11. The system of claim 9, wherein the delivery device in the second state molds to a shape of a surface it is applied to.

12. The system of claim 9, wherein the thermally-reversible delivery device is a hydrogel.

13. The system of claim 9, wherein the delivery device is a dressing.

14. The system of claim 9, wherein the thermally-reversible delivery device includes one or more poloxamers.

15. A treatment system comprising: an aqueous solution comprising: about 25-225 mM of a source of Zn2+, Ag+, or Cu2+ions; about 25-225 mM of a source of an oxidizing agent forming CIO2- ions; about 0.5-10 mM of a succinate; and about 0-5 mM ascorbic acid, the aqueous solution being configured to increase a redox potential at a target site, and a delivery device housing the solution.

16. The system of claim 15, wherein the delivery device is a hydrogel.

17. The system of claim 15, wherein ascorbic acid is present.

18. The system of claim 15, wherein the aqueous solution further includes chlorhexidine gluconate.

19. The system of claim 15, wherein the aqueous solution further includes hyaluronic acid.

20. The system of claim 15, wherein the aqueous solution further includes one or more poloxamers.

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