Plasma activated compositions and methods of use thereof

Abstract

The present invention provides, in some aspects thereof, a plasma-activated liquid composition capable of gelling in situ upon stimulation such as temperature or pH changes or upon induction by light. The plasma-activated liquid compositions are useful in the prophylaxis, treatment and post-treatment of malignant tumors as well as the treatment of various infections. Some aspects of the invention also provide devices, systems, and methods for plasma-activating liquid media and for providing plasma-activated media from liquid compositions as described above.

Description

Field of the Invention

[0001] The present invention relates to plasma-activated compositions and related systems, devices, and methods of use in the prevention, treatment, and post-treatment of various diseases and disorders, particularly infections and cancer.

[0002] background

[0003] In recent years, low-temperature plasma has gained attention due to its use in various medical applications including wound healing, disinfection, and oncology. The mechanism of action of plasma therapy is attributed to the generation of reactive oxygen and nitrogen species (ROS and RNS, respectively, and collectively referred to as RONS) by plasma, which interact with cells to induce subsequent reactions within the cells that can trigger cell signaling cascades (Laroussi, Plasma 2018, 1(1), 47-60; https: / / doi.org / 10.3390 / plasma1010005 To ensure effective treatment, close contact between cells and RONS is required. However, even when localized delivery of plasma is provided by various devices and apparatuses, the gaseous plasma often escapes rapidly from the site of action, thereby reducing the efficacy of the treatment.

[0004] Plasma-activated media (PAMs) have also been used in medical treatments. A medium can be plasma-activated by exposing it to a gas phase, which itself is excited into a plasma, for example, by an electromagnetic (EM) field. ROS and RNS generated in the gaseous plasma dissolve in the medium, thereby plasma-activating it. Advantageously, the plasma-activated medium is most often a liquid—typically a saline or other aqueous solution—because the diffusion of the activated species in liquids is much faster than in solids, and thus the activation duration can be much shorter.

[0005] WO 2015 / 123720 describes a plasma treatment method comprising: providing a plasma source and a screen comprising a hydrogel, and positioning the screen between the plasma source and a surface of a target to be treated with the plasma such that substantially all of the plasma from the plasma source passes through the screen before contacting the surface of the target, and the screen reduces the concentration of one or more species from the plasma; and / or contacting the surface of the target to be treated with a gel composition comprising a gel-forming material and a liquid phase comprising a plasma-activated liquid.

[0006] Labay et al. (Scientific Reports 2019, 9, 16160; https: / / doi.org / 10.1038 / s41598-019-52673-w) studied the production of RONS in alginate hydrogels by comparing two atmospheric pressure plasma jets, KINPen and helium needle, over a range of plasma treatment conditions. The hydrogels demonstrated the ability to sustainably release RONS with cytotoxic potential against bone cancer cells.

[0007] Labay et al. (ACS Appl Mater Interfaces 2020, 12(42), 47256-47269; doi:10.1021 / acsami.0c12930) studied the formation and release of RONS produced in 2% gelatin. In vitro studies of osteosarcoma (SaOS-2) cell lines exposed to plasma-treated gelatin resulted in a time-dependent increase in cytotoxicity with increasing plasma treatment time of gelatin. Although SaOS-2 cell viability decreased to 12%-23% after 72h for cells exposed to 3min treated gelatin, the viability of healthy cells (hMSCs) was retained (~90%), confirming the selectivity of plasma-treated gelatin for cancer cells.

[0008] WO 2021 / 255179 describes a composition comprising an aqueous polymer solution, a bioceramic material and reactive oxygen and nitrogen species (RONS) and their use for the treatment of bone cancer and / or bone tissue regeneration.

[0009] Overview

[0010] Due to the short lifetime of RONS generated by cold plasma devices in the gas phase, there is a great unmet need for compositions with increased residence time at the site of action to induce effective release of RONS for the treatment of various diseases and disorders.

[0011] The present invention provides a composition in liquid form that has been plasma-activated to produce RONS, comprising a synthetic biocompatible polymer capable of in situ gelation in response to a stimulus. The composition can be used for the prevention, treatment, and post-treatment of cancer, as well as for the treatment of viral, bacterial, yeast, mold, and fungal infections, and other medical conditions.

[0012] The present invention is based in part on the unexpected discovery that administration of a plasma-activated liquid that undergoes an in situ phase transition to form a hydrogel has therapeutic efficacy in treating premalignant lesions and preventing tumor recurrence following tumor resection. The compositions of the present invention may also be used to treat various types of cancer, particularly including cancers in soft tissue. Also within the scope of the present invention is the treatment of infections by topical application of a plasma-activated liquid composition, wherein the in situ gel formation provides an extended residence time of the active substance at the site of action, resulting in improved therapeutic efficacy.

[0013] According to a first aspect, there is provided a liquid composition comprising more than one reactive oxygen and nitrogen species generated by a plasma and a synthetic biocompatible polymer, wherein the composition undergoes a phase transition to a gel form upon stimulation.

[0014] In one embodiment, the more than one reactive oxygen and nitrogen species generated by the plasma include H2O2, OH * HO2 * 、O2 - 、O3、NO * 、NO2 - 、NO3 - and ONOO * Each possibility represents a separate embodiment.

[0015] In other embodiments, the synthetic biocompatible polymer comprises polyethylene glycol (PEG), polypropylene glycol (PPG), poly(meth)acrylic acid, poly(meth)acrylate, or a combination thereof. Each possibility represents a separate embodiment. In a specific embodiment, the synthetic biocompatible polymer comprises a poloxamer. In another embodiment, the synthetic biocompatible polymer comprises a poloxamer selected from the group consisting of: poloxamer 101, poloxamer 105, poloxamer 108, poloxamer 122, poloxamer 123, poloxamer 124, poloxamer 181, poloxamer 182, poloxamer 183, poloxamer 184, poloxamer 185, poloxamer 188, poloxamer 212, poloxamer 215, Poloxamer 217, Poloxamer 231, Poloxamer 234, Poloxamer 235, Poloxamer 237, Poloxamer 238, Poloxamer 282, Poloxamer 284, Poloxamer 288, Poloxamer 331, Poloxamer 333, Poloxamer 334, Poloxamer 335, Poloxamer 338, Poloxamer 401, Poloxamer 402, Poloxamer 403, and Poloxamer 407, or mixtures thereof. Each possibility represents a separate embodiment.

[0016] In certain embodiments, the synthetic biocompatible polymer comprises a poloxamer selected from the group consisting of poloxamer 407, poloxamer 188, and poloxamer 338, or mixtures thereof. Each possibility represents a separate embodiment. In specific embodiments, the synthetic biocompatible polymer may comprise a mixture of poloxamers. In certain embodiments, the synthetic biocompatible polymer comprises poloxamer 407 or a mixture of poloxamer 407 and poloxamer 188. In other embodiments, the synthetic biocompatible polymer comprises a mixture of poloxamer 407 and poloxamer 188 in a weight ratio of 10:1 to 1:1, including all iterations of the ratio within the specified range. In other embodiments, the synthetic biocompatible polymer comprises a mixture of poloxamer 407 and poloxamer 188 in a weight ratio of 6:1 to 1:1, including all iterations of the ratio within the specified range. In other embodiments, the synthetic biocompatible polymer comprises a mixture of poloxamer 338 and poloxamer 188. In a specific embodiment, the synthetic biocompatible polymer comprises a mixture of poloxamer 338 and poloxamer 188 in a weight ratio of 10:1 to 1:1, including all iterations of ratios within the specified range. In yet other specific embodiments, the synthetic biocompatible polymer comprises a mixture of poloxamer 338 and poloxamer 188 in a weight ratio of 6:1 to 1:1, including all iterations of ratios within the specified range.

[0017] In various embodiments, the stimulus comprises a temperature change. In other embodiments, the stimulus comprises a pH change. In yet other embodiments, the stimulus comprises light-induced cross-linking.

[0018] In other embodiments, the composition further comprises a fluid medium. In one embodiment, the fluid medium comprises a buffer or a pH adjusting agent. In various embodiments, the buffer or pH adjusting agent is selected from the group consisting of: 2-amino-2-hydroxymethyl-1,3-propanediol (Tris), 2-[bis(2-hydroxyethyl)imino]-2-(hydroxymethyl)-1,3-propanediol (bis-Tris), 4-morpholineethanesulfonic acid (MES) buffer, ammonium chloride, N,N-dihydroxyethylglycine (bicine), N-[tris(hydroxymethyl)methyl]glycine (tricine), sodium dihydrogen phosphate, disodium hydrogen phosphate, sodium carbonate, sodium bicarbonate, sodium acetate, sodium phosphate, glutamic acid, citrate Buffer, histidine buffer, Dulbecco's phosphate buffered saline, 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES), methoxypsoralen (MOPS), N-cyclohexyl-3-aminopropanesulfonic acid (CAPS), N-cyclohexyl-2-hydroxy-3-aminopropanesulfonic acid (CAPSO), N-cyclohexyl-2-aminoethanesulfonic acid (CHES), 3-[4-(2-hydroxyethyl)-1-piperazinyl]propanesulfonic acid (HEPPS), phosphate buffered saline, tris buffered saline, Hank's solution and Ringer's solution, and mixtures or combinations thereof. Each possibility represents a separate embodiment. In another embodiment, the composition has a pH in the range of about 5.0 to about 7.5, including each value within the specified range.

[0019] In various embodiments, the fluid medium comprises a solvent. In particular embodiments, the solvent is an aqueous solvent. In specific embodiments, the fluid medium comprises water.

[0020] In certain embodiments, the compositions disclosed herein can be used to treat premalignant lesions. Thus, according to these embodiments, a method of treating a premalignant lesion in a subject in need thereof is provided, the method comprising contacting the lesion with a therapeutically effective amount of a liquid composition disclosed herein or a therapeutically effective amount of a composition disclosed herein in gel form.

[0021] In other embodiments, the compositions disclosed herein can be used to treat tumors. Thus, according to these embodiments, a method of treating a tumor in a subject in need thereof is provided, comprising contacting the tumor with a therapeutically effective amount of a liquid composition disclosed herein or a therapeutically effective amount of a composition disclosed herein in gel form.

[0022] In other embodiments, the compositions disclosed herein can be used to prevent or delay tumor recurrence after tumor resection. Thus, according to these embodiments, a method of preventing or delaying tumor recurrence after tumor resection in a subject in need thereof is provided, the method comprising contacting the remaining tissue surrounding the tumor with a therapeutically effective amount of a liquid composition disclosed herein or a therapeutically effective amount of a composition disclosed herein in gel form.

[0023] In some embodiments, the compositions disclosed herein can be used to treat viral infections, bacterial infections, yeast infections, mold infections, or fungal infections. Each possibility represents a separate embodiment. Thus, according to these embodiments, a method of treating an infection in a subject in need thereof is provided, the infection being selected from bacterial infections, viral infections, yeast infections, mold infections, and fungal infections, the method comprising topically administering to the subject a therapeutically effective amount of a liquid composition disclosed herein or a therapeutically effective amount of a composition disclosed herein in gel form.

[0024] According to another aspect of the present invention, a system for providing a plasma-activated medium (PAM) for medical use is provided. The system includes a container configured to hold a liquid medium. The system also includes an electrode electrically connected to a high-voltage power supply, the electrode configured to apply an electromagnetic field within the container to generate plasma. The system also includes an actuator configured to cause agitation of the liquid medium within the container. The system also includes a refrigerator having a cold portion, the refrigerator configured to be thermally coupled to the liquid medium within the container. According to aspects of the present invention, the system is configured to generate plasma within the container at atmospheric pressure while simultaneously agitating the liquid medium and maintaining it at a temperature below room temperature.

[0025] It should be noted that the term "chiller" is used herein in a broad sense and refers to any type of cooling device, cooling system, or cooling method.

[0026] According to some embodiments, the liquid medium is a liquid composition comprising a biocompatible polymer, the liquid composition being capable of undergoing a phase transition to a gel form upon stimulation, as described above. According to some embodiments, the biocompatible polymer is synthetic.

[0027] According to another aspect of the present invention, a transportable sealed container is provided. The sealed container includes a liquid medium stored in the container, intended for medical use, and electrodes electrically connected to a high-voltage (HV) connector. The electrodes are configured to apply an EM field within the container that generates a plasma when a high voltage is received via the HV connector. The container is bio-sealed to prevent bacteria or viruses from penetrating the container. According to some embodiments, the container is hermetically sealed.

[0028] According to some embodiments, the liquid medium is sterile. According to some embodiments, the liquid medium comprises a biocompatible polymer forming a liquid composition capable of undergoing a phase transition to a gel form upon stimulation, as described above.

[0029] According to aspects of the present invention, a portable, passive cooler configured to refrigerate a container is also provided. The passive cooler includes a cool pack having at least one chamber containing a coolant. The coolant preferably has a freezing temperature between -10°C and +5°C. The at least one chamber of the cool pack is shaped to surround a slot in the cool pack, wherein the slot is configured and sized to receive the container therein.

[0030] According to another aspect of the present invention, there is provided a foam comprising reactive substances H2O2 and NO2 - and the reactive species OH * HO2 * 、O2 - 、O3、NO * 、NO3 - and ONOO * According to some embodiments, the foam is made from a liquid composition as described above that is activated by plasma and converted into foam by intense agitation.

[0031] From the detailed description given below, additional embodiments and the full scope of applicability of the present invention will become apparent. However, it should be understood that the detailed description and specific examples, while indicating preferred embodiments of the present invention, are given by way of illustration only, as various changes and modifications within the spirit and scope of the present invention will become apparent to those skilled in the art based on this detailed description. BRIEF DESCRIPTION OF THE DRAWINGS

[0033] Figure 1A-Figure 1C Embodiments of a capsule configured to contain a liquid composition therein and a corresponding activation unit are schematically depicted.

[0034] Figure 1D An embodiment of an apparatus for plasma activation of a liquid medium held in a tank is schematically depicted.

[0035] Figure 1E An embodiment of an inlet tube and a hollow needle serving as an electrode is schematically depicted.

[0036] Figure 2A Another embodiment of an apparatus for plasma activation of a liquid medium in a tank is depicted.

[0037] Figure 2B Yet another embodiment of an apparatus for plasma activation of a liquid medium in a tank is schematically depicted.

[0038] Figure 3A An embodiment of a sealed canister containing a liquid medium according to the teachings herein is schematically illustrated.

[0039] Figure 3B Schematic diagram of the plasma activation process Figure 3A Embodiment of the device of the liquid medium in the tank.

[0040] Figure 4A An embodiment of a passive cooler is illustrated that includes an ice pack in the form of a torus surrounding the canister.

[0041] Figure 4B is an exploded view of an embodiment of a passive cooler including an ice pack having an outer chamber and an inner chamber surrounded by the outer chamber.

[0042] Figure 4C An embodiment of an apparatus for plasma activation of a liquid medium incorporating the passive cooler of the present invention is illustrated.

[0043] Figure 5 Shown are images of agar plates on which secretions from infected ears were incubated and treated with a plasma-activated composition (left) or a control (right).

[0044] Figure 6 An embodiment of an endoscope for delivering a plasma-activated composition in liquid form to a cyst in a patient's pancreas is schematically depicted.

[0045] Details

[0046] The present invention provides compositions containing RONS that can be used to treat various diseases and disorders. The compositions are advantageously administered in liquid form and are capable of undergoing a phase transition to a gel form in response to a stimulus, such as upon contact with a site of action. This provides increased residence time at the site of action, thereby enhancing therapeutic efficacy.

[0047] Disclosed herein for the first time are compositions comprising plasma-generated RONS, wherein the compositions gel in situ, i.e., at the site of action, to provide controlled release of the RONS for a prolonged, long, or extended duration. The compositions are highly effective in treating premalignant conditions, malignant tumors, and preventing recurrence of malignant tumors following surgical resection. The compositions can also be used to treat viral, bacterial, yeast, mold, or fungal infections, particularly infections of the ear or skin.

[0048] According to the principles of the present invention, the liquid compositions disclosed herein undergo an in situ phase transition to a gel form in response to a stimulus. As used herein, the term "phase transition" refers to a liquid-gel phase transition and is intended to encompass an increase in the viscosity or hardness of the composition in order to provide a longer residence time at the site of action. It should be understood that the transition can be reversible or irreversible, with each possibility representing a separate embodiment.

[0049] Thus, provided herein is a composition that has been plasma activated to contain more than one reactive oxygen and nitrogen species. In accordance with the principles of the present invention, the composition contains more than one charged ion, free radical, and electron, with a net charge that is substantially neutral. As used herein, the term "reactive oxygen and nitrogen species" or "RONS" refers to a reactive moiety containing oxygen, nitrogen, or both. Typically, the reactive moiety is characterized by a short half-life. Included within this term are moieties such as, but not limited to, H2O2 (hydrogen peroxide), OH * (hydroxyl radical), HO2 * (hydroperoxyl radical), O2 - (superoxide radicals), O3 (ozone), NO * (nitric oxide), NO2 - (nitrogen dioxide), NO3 - (nitrate) and ONOO * (Peroxynitrite). Each possibility represents a separate embodiment.

[0050] According to the teachings herein, the plasma is preferably (but not necessarily) generated in an arc mode at ambient pressure. The arc mode is characterized by a volatile filamentary discharge and has been found by the inventors to be very effective in producing the reactive species described herein. It has been found that the reactive species H2O2, NO2 - and NO3 - Typical production rates of RONS are 10 mg / L, 60 mg / L, and 300 mg / L, respectively, in 5 cc of the liquid composition of the present invention within 2.5 minutes. Typically, the amount of RONS in the composition ranges from about 1 mg / L to about 2,000 mg / L, including each value within the specified range. Exemplary ranges include, but are not limited to, about 10 mg / L to about 1,500 mg / L, about 50 mg / L to about 1,200 mg / L, or about 100 mg / L to about 1,000 mg / L, including each value within the specified range. Each possibility represents a separate embodiment.

[0051] Liquids, particularly liquid compositions comprising polymers, that are configured to transform into gels when exposed to a stimulus can be plasma activated by various methods. Any method capable of plasma activation of a liquid is contemplated in the context of the present invention. Plasma activation is intended to include the generation of a plasma within the medium under consideration (i.e., within the liquid) and, additionally or alternatively, in a gaseous medium that is in contact with or brought into contact with the medium. Furthermore, in order to be considered 'plasma activated' in the context of the present invention, the medium must include, as a result of the plasma activation process as described above, excited species that are not present in the medium in the absence of activation, such as ROS and RNS. Explicit examples of plasma activation of liquids by exposing the liquid to a plasma-excited gas are provided in the Examples below, particularly in Examples 2 to 7.

[0052] According to the principles of the present invention, the composition comprises a biocompatible polymer capable of gelling in response to a stimulus. Thus, the composition is configured to undergo a phase transition to a gel form upon stimulation. In some aspects and embodiments, the phase transition is reversible.

[0053] In certain aspects and embodiments, the phase transition is induced by a change in temperature. In some embodiments, the phase transition can be induced by a decrease in temperature. Thus, for example, a composition in liquid form can be plasma activated at a temperature above body temperature. Then, upon application of the activated composition to a body part and upon reaching body temperature or even below body temperature, the composition can be converted into a gel.

[0054] However, the phase transition from liquid form to gel is generally gradual (as a function of temperature). That is, there can be a significant temperature difference between the gel phase and the liquid phase, for example 10, 15, or even 20 degrees; therefore, to ensure low viscosity during plasma treatment, the composition must be at a temperature significantly above body temperature to ensure gelation. Therefore, a potential disadvantage of this method is that applying the composition to a subject at a temperature when it is a liquid, i.e., a temperature significantly above body temperature, may cause discomfort, pain, or even actual damage.

[0055] In some embodiments, the phase transition can be induced by an increase in temperature. For example, a phase transition can be induced when a temperature of 15°C or higher is reached. In other embodiments, the phase transition is induced when a temperature of 20°C or higher is reached. In further embodiments, the phase transition is induced when a temperature of 25°C or higher is reached. Currently preferred is a phase transition that occurs upon contact with tissue at physiological body temperature. Therefore, within the scope of the present invention are compositions comprising thermoreversible gel-forming polymers that are liquid at low temperatures and are capable of undergoing a phase transition to a gel form at body temperature, wherein they are designed to exert their therapeutic effect, thereby allowing the RONS to be slowly released at a selected site.

[0056] According to some embodiments, the phase transition can be induced by a change in acidity (pH). In some embodiments, the liquid composition of the present invention can gel upon mixing with an aqueous solution such that its acidity (or alkalinity) changes by more than 1 over the pH range.

[0057] According to some embodiments, the liquid compositions of the present invention can be gelled by light-induced cross-linking, as described in further detail below.

[0058] In accordance with the principles of the present invention, the polymer used in the composition is a biocompatible polymer. In some aspects and embodiments, the polymer is biodegradable.

[0059] Within the scope of the present invention are synthetic polymers as well as natural polymers, with each possibility representing a separate embodiment. Suitable polymers include, but are not limited to, polyanhydrides; poly(sebacic acid), SA; poly(ricinoleic acid), RA; poly(fumaric acid), FA; poly(fatty acid dimer), FAD; poly(terephthalic acid), TA; poly(isophthalic acid), IPA; poly(p-{carboxyphenoxy}methane), CPM; poly(p-{carboxyphenoxy}propane), CPP; poly(p-{carboxyphenoxy}hexane), CPH; polyamines, polyurethanes, polyesteramides, polyorthoesters {CHD M: cis / trans-cyclohexanedimethanol, HD: 1,6-hexanediol, DETOU: (3,9-diethylene-2,4,8,10-tetraoxaspiroundecane)}; polydioxanone; polyhydroxybutyrate; polyalkylene oxalate; polyamide; polyesteramide; polyacetal; polyketal; polycarbonate; polyorthocarbonate; polysiloxane; polyphosphazene; succinate; hyaluronic acid; poly(malic acid); poly(amino acid); polyhydroxyvalerate; polyalkylene succinate; polyvinylpyrrolidone Pyrrolidone; polystyrene; synthetic cellulose esters; polyacrylic acid; polybutyric acid; triblock copolymers (PLGA-PEG-PLGA), triblock copolymers (PEG-PLGA-PEG), poly(N-isopropylacrylamide) (PNIPAAm), poly(ethylene oxide)-poly(propylene oxide)-poly(ethylene oxide) triblock copolymers (PEO-PPO-PEO), polyvaleric acid; polyethylene glycol; polyhydroxyalkyl cellulose; chitin; chitosan; polyorthoesters and copolymers, terpolymers; lipids such as cholesterol, lecithin; poly(glutamic acid-co-ethyl glutamate), poly(D,L-lactide-co-glycolide) (PLGA), poly(D,L-lactide) (PLA), polyglycolide (PGA), polycaprolactone (PCL), polyhydroxybutyrate, polyorthoesters, polyalkaneanhydride, gelatin, alginate, collagen, oxidized cellulose, polyphosphazene, and any combination thereof. Each possibility represents a separate embodiment.

[0060] According to the principles of the present invention, preferred polymers are synthetic biocompatible polymers at present. Not subject to the constraint of any theory or mechanism of action, it is expected that the use of synthetic polymers provides, without compromising biocompatibility, the means of producing biocompatible hydrogels with controlled physical properties such as density, hardness and proteolytic degradability by universal synthetic components. The other possible advantage of using synthetic biocompatible polymers within the scope of the present invention can be the reversibility of phase transition, which avoids the use of cross-linking agents and is controlled by the stimulation present in physiological conditions. In some embodiments, the polymer used in the compositions disclosed herein is not a natural polymer, such as alginate or gelatin.

[0061] Within the scope of the present invention are synthetic biocompatible polymers comprising polyoxyethylene-polyoxypropylene block copolymers known as "poloxamers." Exemplary and non-limiting poloxamers include poloxamer 101, poloxamer 105, poloxamer 108, poloxamer 122, poloxamer 123, poloxamer 124, poloxamer 181, poloxamer 182, poloxamer 183, poloxamer 184, poloxamer 185, poloxamer 188, poloxamer 212, poloxamer 215, poloxamer 217, poloxamer 218, poloxamer 219, poloxamer 220, poloxamer 221, poloxamer 222, poloxamer 223, poloxamer 224, poloxamer 225, poloxamer 226, poloxamer 227, poloxamer 228, poloxamer 229, poloxamer 230, poloxamer 231, poloxamer 232, poloxamer 233, poloxamer 234, poloxamer 235, poloxamer 236, poloxamer 237, poloxamer 238, poloxamer 239, poloxamer 240, poloxamer 241, poloxamer 242, poloxamer 243, poloxamer 244, poloxamer 245, poloxamer 246, poloxamer 247, poloxamer 248, poloxamer 249, poloxamer 250, poloxamer 251, poloxamer 252, poloxamer 253, poloxamer Preferably, poloxamer 231, poloxamer 234, poloxamer 235, poloxamer 237, poloxamer 238, poloxamer 282, poloxamer 284, poloxamer 288, poloxamer 331, poloxamer 333, poloxamer 334, poloxamer 335, poloxamer 338, poloxamer 401, poloxamer 402, poloxamer 403, and poloxamer 407 or a mixture thereof. Each possibility represents a separate embodiment. Currently preferred are poloxamer 407, poloxamer 188, and poloxamer 338 or a mixture thereof. Each possibility represents a separate embodiment.

[0062] In certain aspects and embodiments, the composition comprises a mixture of poloxamer 407 and poloxamer 188. Suitable weight ratios of poloxamer 407 and poloxamer 188 include, but are not limited to, ratios of 10:1 to 1:1, including any ratios therebetween. Currently preferred is a ratio of 6:1 to 1:1, such as 4:1.

[0063] In specific aspects and embodiments, the composition comprises a mixture of poloxamer 338 and poloxamer 188. Suitable weight ratios of poloxamer 338 to poloxamer 188 include, but are not limited to, ratios of 10:1 to 1:1, including any ratios therebetween. Currently preferred is a ratio of 6:1 to 1:1, such as 4:1.

[0064] In various aspects and embodiments, the phase transition is induced by a change in pH, for example, when an acidic pH or a neutral to alkaline pH is reached. Each possibility represents a separate embodiment. To achieve a phase transition in response to pH, the composition typically contains a pH-responsive polymer, such as an enteric polymer or a reverse enteric polymer. Each possibility represents a separate embodiment.

[0065] The term "enteric polymer" as used herein refers to a polymer characterized by increased permeability at pH values ​​(e.g., intestinal fluid) above pH 5.0 while remaining insoluble at low pH values ​​(such as those found in the gastric environment). Exemplary and non-limiting enteric polymers include acrylic acid and (meth) acrylic acid copolymers, polyvinyl acetate phthalate, and the like. Each possibility represents a separate embodiment. Acrylic acid and methacrylic acid copolymers are anionic copolymers based on (meth) acrylic acid and (meth) acrylic acid alkyl esters, such as, but not limited to, polymethacrylic acid, polymethyl methacrylate, polyethyl methacrylate, and polyethyl acrylate. Commercial acrylic acid and methacrylic acid copolymers are available under the trade name Eudragit (Evonik Industries AG, Essen, Germany) and are typically provided as powders or aqueous dispersions, including, but not limited to, Eudragit L 30D-55, Eudragit L 100-55, Eudragit L 100, Eudragit L 12.5, Eudragit NE 40D, Eudragit RL 100, Eudragit S100, Eudragit S12.5, Eudragit FS 30D, Eudragit RLPO, Eudragit RL 12.5, Eudragit RL 30D, Eudragit RS100, Eudragit RS PO, Eudragit RS 30D, Eudragit RS12.5, Eudragit NE 30D, Eudragit NM 30D, or combinations and mixtures thereof. Each possibility represents a separate embodiment.

[0066] As used herein, " enteric polymers" refer to polymers that are insoluble at a pH greater than that found in the stomach, i.e., insoluble at a pH greater than 5.0 but soluble at an acidic pH. Exemplary enteric polymers include, but are not limited to, (meth) acrylate polymers or copolymers, such as acrylate and methacrylate copolymers with primary, secondary, or tertiary amino groups or quaternary ammonium groups. These enteric polymers are commercially available as Eudragit E 100, Eudragit E 12.5, Eudragit EPO, Eudragit RL 100, or combinations and mixtures thereof. Each possibility represents a separate embodiment.

[0067] Typically, the amount of the biocompatible polymer synthesized in the composition is in the range of from about 1% to about 50% of the total weight of the composition, including each value within the specified range. Currently preferred amounts include those that are greater than 5% of the total weight of the composition. Exemplary amounts include, but are not limited to, about 1%, about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45% or about 50% of the total weight of the composition, wherein each possibility represents a separate embodiment.

[0068] According to various aspects and embodiments, the biocompatible polymer is dispersed or dissolved in a fluid medium, which may further include a buffer or a pH regulator and a solvent. According to the principles of the present invention, one or more buffers or pH regulators may be included in the composition. Such buffers or pH regulators include, but are not limited to, 2-amino-2-hydroxymethyl-1,3-propanediol (Tris), 2-[bis(2-hydroxyethyl)imino]-2-(hydroxymethyl)-1,3-propanediol (bis-Tris), 4-morpholineethanesulfonic acid (MES) buffer, ammonium chloride, N,N-dihydroxyethylglycine, N-[tris(hydroxymethyl)methyl]glycine, sodium dihydrogen phosphate, disodium hydrogen phosphate, sodium carbonate, sodium bicarbonate, sodium acetate, sodium phosphate, glutamic acid, citrate buffer, histidine buffer,

[0014] The invention relates to a novel polysaccharide comprising: phosphate-buffered saline, 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES), methoxypsoralen (MOPS), N-cyclohexyl-3-aminopropanesulfonic acid (CAPS), N-cyclohexyl-2-hydroxy-3-aminopropanesulfonic acid (CAPSO), N-cyclohexyl-2-aminoethanesulfonic acid (CHES), 3-[4-(2-hydroxyethyl)-1-piperazinyl]propanesulfonic acid (HEPPS), phosphate-buffered saline, tris-buffered saline, Hank's solution, and Ringer's solution, or mixtures or combinations thereof. Each possibility represents a separate embodiment.

[0069] According to various aspects and embodiments, the composition has a pH in the range of about 5.0 to about 7.5, including each value within the specified range. Exemplary non-limiting ranges include about 5.3 to about 6.3, about 5.8 to about 6.8, about 6.0 to about 7.0, etc., including each value within the specified range. For example, the composition of the present invention can have a pH of about 5.0, about 5.1, about 5.2, about 5.3, about 5.4, about 5.5, about 5.6, about 5.7, about 5.8, about 5.9, about 6.0, about 6.1, about 6.2, about 6.3, about 6.4, about 6.5, about 6.6, about 6.7, about 6.8, about 6.9, about 7.0, about 7.1, about 7.2, about 7.3, about 7.4 or about 7.5, wherein each possibility represents a separate embodiment.

[0070] In various aspects and embodiments, the fluid medium of dispersion or dissolving polymer is the aqueous medium comprising water or salt solution as solvent.Usually, the amount of solvent comprises each value in the specified range from about 50% to about 99% scope of the gross weight of composition.Exemplary amount includes but is not limited to about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95% or about 99% of the gross weight of composition, wherein every kind of possibility represents independent embodiment.

[0071] In some aspects and embodiments, the composition further comprises an ionic tonicity agent, such as sodium chloride, or a non-ionic tonicity agent, such as a polyol.Additional excipients that may be present in the composition include, but are not limited to, surfactants and preservatives as known in the art.

[0072] According to the principles of the present invention, the compositions disclosed herein form gels (hydrogels) upon stimulation such as changes in temperature or pH or light-induced crosslinking. The term "hydrogel" according to the present invention refers to a three-dimensional hydrated assembly of biocompatible nanofibers. The liquid-gel phase transition can be characterized by an increase in the viscosity of the material, wherein the increase is at least one order of magnitude in a range of 20°C or less. Typically, the gel is characterized by a viscosity greater than 20,000 mPa·s, preferably greater than 50,000 mPa·s, and even more preferably greater than 100,000 mPa·s. Each possibility represents a separate embodiment. In the liquid phase, the composition is typically characterized by a viscosity less than 600 mPa·s, preferably less than 300 mPa·s, and even more preferably less than 100 mPa·s. Each possibility represents a separate embodiment. Viscosity measurements can be performed as known in the art, for example using a suitable viscometer, including but not limited to a Brookfield viscometer or an Anton Paar RheoPlus viscometer with appropriate settings. It was commented that by determining the viscosity of the gel phase, the temporal profile of the RONS release rate can be determined, whereby the greater the viscosity, the slower the release.

[0073] In certain aspects and embodiments, the compositions disclosed herein can be used for the prevention, treatment, and post-treatment of cancer. As used herein, the term "cancer" refers to a disorder in which a cell population becomes, to varying degrees, unresponsive to the control mechanisms that normally govern proliferation and differentiation. Cancer refers to various types of malignant neoplasms and tumors, including primary tumors and tumor metastases. Non-limiting examples of cancers that can be treated by the compositions of the present invention are brain cancer, ovarian cancer, colon cancer, prostate cancer, kidney cancer, bladder cancer, breast cancer, lung cancer, oral cancer, and skin cancer. Each possibility represents a separate embodiment. In one embodiment, the cancer is a soft tissue cancer. Specific examples of cancer include, but are not limited to, carcinomas, sarcomas, myelomas, leukemias, lymphomas, and mixed tumors. Each possibility represents a separate embodiment. Additional examples of cancer include, but are not limited to, lymphoproliferative disorders, breast cancer, ovarian cancer, prostate cancer, cervical cancer, endometrial cancer, liver cancer, bladder cancer, stomach cancer, colon cancer, colorectal cancer, pancreatic cancer, thyroid cancer, esophageal cancer, head and neck cancer, central nervous system cancer, peripheral nervous system cancer, skin cancer, kidney cancer, and metastases of all of the above. Each possibility represents a separate embodiment. Specific types of cancer include, but are not limited to, hepatocellular carcinoma, hepatoma, hepatoblastoma, rhabdomyosarcoma, esophageal cancer, thyroid cancer, ganglioblastoma, fibrosarcoma, myxosarcoma, liposarcoma, chondrosarcoma, osteosarcoma, chordoma, angiosarcoma, endotheliosarcoma, Ewing's tumor, leiomyosarcoma, rhabdotheliosarcoma, invasive ductal carcinoma, papillary adenocarcinoma, melanoma, squamous cell carcinoma, basal cell carcinoma, adenocarcinoma (well differentiated, moderately differentiated, poorly differentiated, or undifferentiated) and ulcerative colitis. The invention also includes but is not limited to: chemokines, cytokines, cytokines, leukemia, multiple myeloma, myeloid lymphoma, Hodgkin lymphoma, non-Hodgkin lymphoma, and hepatocarcinoma. Each possibility represents a separate embodiment.

[0074] In some representative embodiments, the cancer is selected from the group consisting of colorectal cancer and bladder cancer.Each possibility represents a separate embodiment.

[0075] In the context of the present invention, the term "treatment of cancer" includes at least one of the following: a reduction in the growth rate of the cancer (i.e., the cancer is still growing, but at a slower rate); an arrest of the growth of the cancerous growth, i.e., tumor growth stagnation, and in preferred cases, a reduction or reduction in the size of the tumor. The term also includes a reduction in the number of metastases, a reduction in the number of new metastases formed, a slowing of the progression of cancer from one stage to another, and a reduction in angiogenesis induced by cancer. In the most preferred case, the tumor is completely eliminated. The term additionally includes prolonging the survival of subjects undergoing treatment, prolonging the time to disease progression, tumor regression, and the like. It should be understood that the term "treating cancer" also refers to inhibiting the proliferation of malignant (cancer) cells, including tumor formation, primary tumors, tumor progression, or tumor metastasis. The term "inhibit proliferation" with respect to cancer cells may also refer to at least one of the following: a decrease in cell number (due to cell death, which may be necrosis, apoptosis, or any other type of cell death, or a combination thereof) compared to a control; a decrease in cell growth rate, i.e., the total number of cells may increase, but at a lower level or at a lower rate compared to the increase in the control; a decrease in the invasiveness of the cells (as determined, for example, by a soft agar assay), even if their total number does not change, compared to a control; progression from a less differentiated cell type to a more differentiated cell type; a deceleration of neoplastic transformation; or alternatively, a slowing of the progression of cancer cells from one stage to the next. Each possibility represents a separate embodiment.

[0076] The specific groups for which the compositions of the present invention are beneficial include subjects with premalignant tumors. Therefore, the use of the compositions disclosed herein in the preparation of a medicament for treating premalignant lesions is provided. As used herein, the term "premalignant" refers to cysts, polyps, lesions, etc., or any form of cell structure or growth disorder that is not malignant but has an increased probability of canceration. In particular, the term refers to tissues that exert one or more of the following: morphological or structural changes, histological changes showing cell atypia or dysplasia, and initial molecular changes in gene expression or expression of specific isoforms of proteins. Each possibility represents a separate embodiment. Specific types of premalignant tumors include, but are not limited to, myelodysplastic disorders, cervical carcinoma in situ, familial intestinal polyposis (e.g., Gardner syndrome), oral leukoplakia, histiocytosis, keloids, hemangiomas, hyperkeratosis, and papulosquamous rashes. Each possibility represents a separate embodiment. Non-cancerous hyperproliferative diseases such as warts and psoriasis are also included. Each possibility represents a separate embodiment.

[0077] Another patient population suitable for treatment with the compositions disclosed herein is a subject undergoing tumor resection to prevent or delay tumor recurrence. Therefore, the use of a composition as disclosed herein in the preparation of a medicament for preventing or delaying tumor recurrence after tumor resection is provided. Cancer recurrence after resection is a known problem in many cancer types, often attributed to failure to remove all cancer cells during surgery, spread of cancer cells during percutaneous ablation, and resistance to chemotherapeutic agents. The compositions disclosed herein are particularly beneficial in preventing or delaying tumor recurrence after tumor resection due to their increased residence time at the application site.

[0078] Within the scope of the present invention is a liquid composition comprising more than one reactive oxygen and nitrogen species generated by a plasma, for use in preventing or delaying tumor recurrence following tumor resection. Thus, according to these embodiments, a method of preventing or delaying tumor recurrence following tumor resection in a subject in need thereof is provided, the method comprising contacting the remaining tissue surrounding the tumor with a therapeutically effective amount of a liquid composition comprising more than one reactive oxygen and nitrogen species generated by a plasma. In one embodiment, the subject in need thereof suffers from bladder cancer.

[0079] As used herein, the term "contacting" in the above-mentioned treatment context refers to topical administration, i.e., contact with the composition of the present invention. Contacting can be achieved with cells or tissue culture, or with a living organism, such as a human. Thus, the term "contacting" can be ex vivo, on a surface, on a device, in a cell / tissue culture dish, etc., or in vivo, such as in a living organism.

[0080] The present invention encompasses the use of the compositions disclosed herein to treat infections, such as bacterial infections, viral infections, yeast infections, mold infections, and / or fungal infections. As used herein, the term "treating" with respect to infection means stopping or slowing the progression of the disease. The term "treating" also includes reducing the occurrence of various symptoms associated with the infection. In one embodiment, with respect to bacterial or viral infections, treatment includes inhibiting bacterial or viral replication while reducing bacterial or viral load. In other embodiments, treatment includes substantially complete eradication of the infectious disease.

[0081] Specific bacteria types include but are not limited to Gram-positive bacteria and Gram-negative bacteria. Exemplary bacteria include Streptococcus, Staphylococcus, Haemophilus, Moraxella, Micrococcus, Corynebacterium, Clostridium, Pseudomonas, Proteus, Peptostreptococcus, Neisseria and Escherichia. Every possibility represents a separate embodiment.

[0082] Viral infections that can be treated, inhibited, reduced, or suppressed by the compositions disclosed herein include, but are not limited to, herpes virus infections, such as herpes simplex virus (HSV) type 1, HSV type 2, varicella-zoster virus, cytomegalovirus, Epstein-Barr virus (EBV), human herpes virus 6 (variants A and / or B), human herpes virus 7, and human herpes virus 8 (Kaposi's sarcoma-associated herpesvirus); human papillomavirus (HPV); molluscum contagiosum virus (WV), and skin diseases associated with viral infections, such as warts, and benign tumors of the skin and / or mucous membranes caused by papillomaviruses, e.g., plantar warts (Verrucae plantares), common warts (Verrucae vulgaris), juvenile flat warts (Verrucae planae juveniles), epidermodysplasia verruciformis, condylomata acuminata, condylomata plana, Bowenoid papulosis (Bowenoid papulosis), and dermatological diseases. papulosis), papillomas and focal epithelial hyperplasias on the laryngeal and oral mucosa. Each possibility represents a separate embodiment.

[0083] Within the scope of the present invention is the treatment of fungal infections, including infections caused by fungi or yeasts. Infections can occur in the skin, fingernails, toenails, and mucous membranes, including but not limited to the mouth, pharynx, esophagus, lungs, and genitals (including vagina and penis), or can be systemic, for example, in immunocompromised patients. Each possibility represents a separate embodiment.

[0084] Pathogenic yeasts include, but are not limited to, various species of the genus Candida, such as Candida auris, Candida albicans, Candida glabrata, Candida enolase, Candida tropicalis, Candida krusei, Candida parapsilosis, Candida stellatoidea, Candida parakawsei, Candida lusitaniae, Candida pseudotropicalis, Candida viswanathii, and Candida guillainensis. guilliermondii), and species of Aspergillus, Rhizopus, Mucor, Histoplasma, Coccidioides, Blastomyces, Trichophyton, Microsporum, and Epidermophyton, such as, but not limited to, Aspergillus fumigatus, Aspergillus flavus, Aspergillus niger, Histoplasma capsulatum, Coccidioides immitis, and Blastomyces dermatitidis. Each possibility represents a separate embodiment.

[0085] The present invention encompasses infections of the skin, ears, eyes, nose, throat, lungs, gastrointestinal tract, urinary tract, or genitals. Each possibility represents a separate embodiment. In some representative embodiments, the infection is an ear infection known as otitis media. In other representative embodiments, the infection is a skin infection, such as Propionibacterium acne, which causes juvenile acne.

[0086] The compositions disclosed herein are administered in a therapeutically effective amount. A "therapeutic" treatment is a treatment administered to a subject exhibiting pathological signs for the purpose of alleviating or eliminating those signs. A "therapeutically effective amount" is an amount of the plasma-generated moiety sufficient to provide a beneficial effect to the subject to whom the composition is administered. Typically, the subject is a mammal, such as a human, a pet, or any domesticated animal. In some aspects and embodiments, the compositions disclosed herein are intended for veterinary use.

[0087] Determining a therapeutically effective amount is well within the capabilities of those skilled in the art, particularly in light of the disclosure provided herein. The exact amount of RONS can be selected by an individual physician based on the patient's condition. It generally depends on certain parameters of the subject being treated, such as weight, age, and severity of the disease. The composition can be administered in a single dose or in multiple doses, either continuously or intermittently. As used herein, the term "intermittent" refers to stopping and starting at regular or irregular intervals. For example, intermittent administration can be daily administration for a period of time, cyclic administration, or administration every other day. Each possibility represents a separate embodiment.

[0088] The compositions of the present invention are typically plasma activated prior to administration or contact with the site of therapeutic use. Specific examples of methods of plasma activation of the compositions of the present invention are provided in the Examples below.

[0089] According to some embodiments, the composition according to the present invention may comprise a pharmaceutical agent, such as an antibiotic and / or a steroid. Such a pharmaceutical agent may be added to the composition of the present invention during processing, possibly after plasma activation and before the composition is applied to the patient.

[0090] Where a range of values ​​is provided, it is understood that every intervening value between the upper and lower limits of the range, to one-tenth of the unit of the lower limit (unless the context clearly indicates otherwise), and any other stated or intervening value in the stated range, is encompassed within the present invention. The upper and lower limits of these smaller ranges may independently be included in smaller ranges and are also encompassed within the present invention, subject to any specifically excluded limits in the stated range. Where a stated range includes one or both of the limits, ranges excluding either or both of those included limits are also encompassed within the present invention.

[0091] As used herein, the term "about" when combined with a value means ±10% of the reference value, or if the value is in degrees Celsius (expressed herein as °C), the greater of the reference value and ±1 degree Celsius.

[0092] It should be noted that, as used herein and in the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to "a synthetic biocompatible polymer" includes more than one such polymer known to those skilled in the art and equivalents thereof, and so forth. It should also be noted that the claims can be drafted to exclude any optional element. Thus, this statement is intended to serve as antecedent basis for use of exclusive terminology such as "solely," "only," and the like in connection with the recitation of claim elements, or for use of a "negative" limitation.

[0093] In those instances where similar conventions to "at least one of A, B, and C, etc." are used, such construction is generally intended to be in the sense that one skilled in the art would understand in the convention (e.g., "a composition having at least one of A, B, and C" would include, but is not limited to, compositions having A alone, B alone, C alone, A and B together, A and C together, B and C together, and / or A, B, and C together, etc.). One skilled in the art would also understand that any transitional words and / or phrases, whether in the specification, claims, or drawings, that actually represent two or more alternative terms should be understood to contemplate the possibility of including one term, either of the terms, or both terms. For example, the phrase "A or B" would be understood to include the possibility of "A" or "B" or "A and B."

[0094] It should be understood that certain features of the present invention described in the context of separate embodiments for clarity are also provided in combination in a single embodiment. Conversely, various features of the present invention described in the context of a single embodiment for brevity may also be provided individually or in any suitable subcombination. All combinations of embodiments related to the present invention are specifically included in the present invention and disclosed herein, just as each and every combination is individually and clearly disclosed. In addition, all subcombinations of the various embodiments and elements thereof are also specifically included in the present invention and disclosed herein, just as each and every such subcombination is individually and clearly disclosed herein.

[0095] Additional objects, advantages, and novel features of the present invention will become apparent to those skilled in the art upon examination of the following examples, which are not intended to be limiting. Additionally, each of the various embodiments and aspects of the present invention as described above and as claimed in the claims section below finds experimental support in the following examples.

[0096] Various embodiments and aspects of the present invention as delineated hereinabove and as claimed in the claims section below find experimental support in the following examples. Example

[0097] Example 1

[0098] Composition

[0099] A composition comprising a mixture of poloxamers was prepared while continuously cooling and stirring (Table 1). Cold water (5°C) was first poured into the container and then the poloxamers were slowly added while stirring. The composition was homogenized by continuous stirring for 2 hours.

[0100] Table 1.

[0101] quality% gram Poloxamer 407 20 4.0 Poloxamer 188 5 1.0 water 75 15.0 total 100 20.0

[0102] Additional compositions as outlined in Table 2 were prepared in the same manner.

[0103] Table 2.

[0104]

[0105] Example 2

[0106] Treating Ear Infections in Pets

[0107] Otitis externa is inflammation of the external auditory canal distal to the eardrum. It is one of the most common reasons pets, especially dogs, are referred to the veterinarian. Inflammation can be caused by one of several primary causes, such as foreign matter in the ear canal, allergies, parasites, autoimmune disorders, and other causes. However, long-term irritation caused by the primary cause can develop into an infection caused by fungi, yeasts (e.g., Malassezia), or bacteria (Staphylococcus, Streptococcus, Enterococcus, Pseudomonas, Proteus, etc.). Lack of treatment can lead to increased irritation, increased inflammation, pain, and progression of inflammation to the middle ear and damage, leading to hearing loss. The most common treatment involves administering antibiotics and antifungal medications to disinfect the ear. However, the growing reluctance to use antibiotics and the resulting trend toward alternative treatments has fueled the need for plasma-based disinfection therapies.

[0108] Thus, according to some embodiments, a plasma activated medium (PAM) can be applied to the ear of a subject, particularly a pet's ear, to disinfect the ear and reduce or eliminate inflammation. In some embodiments, a liquid composition comprising a synthetic biocompatible polymer capable of in situ gelation can be prepared as described above. The preferred stimulus for gelation can be selected as a temperature increase from typical room temperature (e.g., about 26°C) to typical body temperature (e.g., about 37°C). In some embodiments, the liquid composition can be converted into a gel when heated to above 30°C or above 35°C. The liquid composition can be activated by plasma, as described in detail below. The resulting PAM can then be applied to the ear of the subject, where the composition is converted into a gel and stabilized in situ. Thus, the plasma activated gel can release RONS onto the infected area over a predetermined duration of several hours or days, after which the gel decomposes.

[0109] The liquid compositions of the present invention can be plasma-activated in large quantities at a factory or, alternatively or alternatively, in small quantities at a clinic before application of the activated medium. Factory activation offers the advantage of ease of use for the end user, as clinic practitioners are provided with an activated liquid that can be easily applied. Additional advantages may include product standardization and potential cost reductions due to large-scale processing. A disadvantage of factory plasma activation is that RONS concentration decreases over time, which can lead to reduced efficacy. Therefore, in some embodiments, plasma activation of the composition at a clinic before application may be preferred.

[0110] Regardless of whether the plasma activation is performed in a factory or in a clinic before application, in large or small quantities, the temperature of the activated medium should preferably be kept well below the gelation temperature during activation.

[0111] Figure 1A-Figure 1C The capsule 10 ( Figure 1A ) embodiment, the capsule 10 is configured to contain therein a liquid composition comprising a polymer 12 ( Figure 1B ). The liquid composition may be converted to a gel upon heating, for example, from typical room temperature to typical body temperature. In some embodiments, the liquid composition may be converted to a gel upon heating to above 30°C or above 35°C. The corresponding activation unit 60 ( Figure 1B ) can be used to plasma activate the liquid composition within the capsule. After plasma activation, the capsule can be used as a syringe barrel 14 for applying the activated liquid ( Figure 1C ).

[0112] Figure 1AA semi-exploded view of an embodiment of a capsule 10 is schematically depicted. Capsule 10 includes a syringe barrel 14 having a funnel 16 with an opening 18 at its distal end. The funnel is configured to facilitate application of a liquid composition into a subject's ear after plasma activation. Syringe barrel 14 can be made of a dielectric material such as glass or plastic. In some embodiments, syringe barrel 14 can be made of metal. Capsule 10 also includes a funnel cap 20 that is removable from syringe barrel 14 and configured to cover the funnel and thereby seal funnel opening 18 to prevent leakage of the liquid composition and infiltration of contaminants into the syringe barrel. Funnel cap 20 includes a cap opening 22 at its distal end and a one-way valve 24 that is configured to allow pressurized gas to flow from the syringe barrel and funnel opening 22 to the environment during plasma activation, as explained further below.

[0113] The capsule 10 also includes a curved tube 30 that passes through a cork 32 and extends between a tube port 34 at the cork and a tube distal end 36. The curved tube 30 can comprise a dielectric material such as glass or plastic. A needle electrode 38 is concentrically arranged along the curved tube, has a tip 40 near the tube distal end 36, and, in some embodiments, retracts from the tube distal end toward the interior of the tube.

[0114] During the manufacture and assembly of the capsule 10, the syringe barrel 14 can be filled with a liquid composition comprising the polymer 12 and then sealed at both ends by the funnel cap 20 and the cork 32, respectively. The tube port 34 can also be sealed by a removable seal 38. The removable seal 38 can be a removable covering, such as an adhesive foil, configured to seal the tube port 34 and prevent contaminants from penetrating into the tube 30. It should be noted that the amount of liquid composition within the syringe barrel 14 can be calibrated so that when the capsule 10 is positioned with the funnel 16 pointing upward, as shown in FIG. Figure 1B As depicted, the liquid surface is below the bend of the curved tube 30 and above the distal end 36 of the tube.

[0115] Plasma activation of the liquid composition 12 may be performed using an activation unit 60, such as Figure 1BSchematically depicted in . Activation unit 60 includes a housing 62 having a slot 64, which is sized and configured to receive capsule 10 therein. Activation unit 60 also includes a compressor 66, which is configured to compress air from the environment toward slot port 70. Additionally or alternatively, in some embodiments, a gas other than air can be used. In some embodiments, a pressurized gas source (not shown here) can be used together with or instead of compressor 66 to force the gas toward slot port 70. Such a pressurized gas source can be a gas reservoir containing, for example, an inert gas such as nitrogen (N2), argon or helium (to name a few examples). Therefore, in some embodiments, a plasma of some inert gases can be generated in capsule 10, and in some embodiments, a plasma of a mixture of air and another gas can be generated.

[0116] Activation unit 60 also includes an HV power supply 72, which is configured to generate a high voltage that is sufficient to apply an electric field that generates plasma. The HV power supply can generate a voltage that is generally higher than 10KV or higher than 6KV or higher than 3kV or even higher than 1KV. Each possibility represents a separate embodiment. The HV supplied by the HV power supply 72 can be a constant current (DC) or an alternating current (AC). In some embodiments, the HV can be at a radio frequency (RF) between 3KHz and 10GHz. The HV power supply 72 is electrically connected to the HV connector 74 in the slot 64. In some embodiments, the HV connector is a part of the slot port 70. It should be noted that the term "electromagnetic field" (or EM field) therefore also can include an electrostatic field.

[0117] For plasma activation of the liquid composition, the capsule 10 with the liquid composition 12 inside is placed in the tank 64 of the activation unit 60, as shown in FIG. Figure 1B 30. The tube port 34 is connected to the tank port 70, so that the compressor 66 is in fluid communication with the curved tube 30, and the HV power supply 72 is electrically connected to the electrode 38 via the HV connector 74. In some embodiments, where the syringe barrel 14 is metal, it can be connected to ground potential. When the HV power supply 72 is activated, a plasma-generating electric field is applied between the electrode tip 40 and the liquid composition, which is at ground potential, thereby generating plasma in this space.

[0118] Since the plasma is generated at ambient pressure by an electric field applied between the tip electrode (in the present embodiment, the tip 40) and the liquid, the plasma is generated in an arc mode. The arc mode is characterized by a volatile filamentary discharge and has been found by the inventors to be very effective in generating reactive species as described herein.

[0119] Typically, the compressor 66 is activated in conjunction with the HV power supply, causing air from the environment to be compressed into the curved tube 30 for release via the bubbles 78 and again to the environment via the one-way valve 24. The generation of bubbles in the liquid composition 12, wherein the bubbles are filled with excited air, helps to increase the liquid surface affected by the plasma and agitate and mix the liquid composition, thereby helping to prevent heating of limited portions of the liquid composition and, therefore, prevent undesirable gelation due to localized temperature increases in the liquid.

[0120] According to some embodiments, the funnel cap 20 is only used to tightly seal the funnel opening 18 and does not include a one-way valve. In such an embodiment, the funnel cap 20 can be removed from the capsule after the capsule is placed in the tank 64 but before the plasma is activated, and the air escaping from the curved tube by the air bubble is freely released into the environment through the funnel opening 18.

[0121] After the plasma activation is complete, the capsule 10 can be removed from the slot 64 and taken out for application of the liquid composition in the ear of a subject. While holding the capsule with the barrel funnel 16 pointing downward, the cork 32 can be removed from the barrel 14 and, alternatively, the plunger 80 can be slightly inserted therein. The funnel cap 20 can then be removed, as shown. Figure 1C In this configuration, a syringe comprising a barrel 14 having a plunger 80 can be withdrawn for application of a liquid composition in the ear of a subject.

[0122] Example 3

[0123] Plasma activation of liquid media

[0124] Figure 1D An embodiment of an apparatus 100 for plasma activation of a liquid medium 110 is schematically depicted, the liquid medium being stored in a tank 120. In some embodiments, it may be undesirable to allow the temperature of the liquid medium to rise above a given temperature during plasma activation. In some embodiments, it may be desirable to maintain the temperature of the liquid medium below room temperature. Therefore, in some embodiments, it may be advantageous to apply cooling, either directly or indirectly, to the liquid medium.

[0125] In some embodiments, the liquid medium 110 can be converted into a gel when heated, for example, when heated from typical room temperature to typical body temperature. In some embodiments, the liquid medium can be converted into a gel when heated to above 30°C or above 35°C. The tank 120 is configured to store and possibly transport the liquid medium. The tank 120 can be made of a dielectric material such as glass or plastic. In some embodiments, the tank 120 can be metal. The tank 120 can be closed with a cover (not shown here) that can be used to hermetically seal the tank and should be removed prior to the plasma treatment described herein. Additionally or alternatively, the tank 120 can be hermetically sealed (i.e., sealed to prevent gas from passing through it) or bio-sealed (sealed to prevent bacteria or viruses from penetrating) using a foil seal (not shown here) that is removed or torn open by hand prior to plasma treatment.

[0126] The device 100 includes a housing 130 having a slot 132 sized and configured to receive a canister 120 therein, and a movable door 140 configured to close over the slot. The door 140 includes a seal 142 configured to seal an opening of the canister 120 when the canister 120 is in the slot 132 and the door 140 is closed.

[0127] Apparatus 100 also includes a gas circulation system 150 configured to circulate gas through tank 120 and through the liquid composition during plasma activation. Gas circulation system 150 includes a compressor 152 configured to collect gas from an outlet pipe 154 and compress the gas into an inlet pipe 160. Outlet pipe 154 extends between compressor 152 and outlet pipe distal end 156. Outlet pipe distal end 156 is open-ended and configured to be in fluid communication with the top portion of tank 120 when the tank is in tank 132 and door 140 is closed, thereby serving as a gas outlet port for tank 120. Similarly, inlet pipe 160 extends between compressor 152 and inlet pipe distal end 162. Distal end 162 is open-ended and configured to penetrate into tank 120 and establish fluid communication with the fluid held therein when the tank is in tank 132 and door 140 is closed, thereby serving as a gas inlet port for tank 120.

[0128] The device 100 also includes a high voltage (HV) power supply 170 electrically connected to an electrode 172, the high voltage (HV) power supply 170 being configured to apply an EM field that generates a plasma when supplied with a suitable HV. In some embodiments, the electrode 172 can be a pointed electrode. In some embodiments, the electrode 172 can be a needle as described in the figure. The electrode 172 can be electrically connected to the HV power supply 170 by galvanic contact via a wire 174, but other forms of electrical connection, such as capacitive coupling or inductive coupling, are also contemplated. In some embodiments, where the tank 120 is metal, the tank can be electrically connected to a ground potential, thereby attributing the ground potential to the liquid medium 110 therein.

[0129] The HV power supply 170 can supply a HV that is constant in time (direct current - DC) or alternating current (AC) to the electrode 172. In some embodiments, the HV is at a frequency greater than 3 kHz (referred to as RF). In some embodiments, the HV is at a frequency between 30 kHz and 10 GHz, including each value within the specified range. In some embodiments, plasma 178 is generated between the electrode and the liquid medium in an arc mode. In some embodiments, plasma is generated in the space above the liquid medium. In some embodiments, the EM field that generates plasma is applied between the electrode 172 and another component of the device 100 (e.g., the wall of the tank 120). In some embodiments, plasma is generated between two electrodes (not shown here), neither of which is pointed; for example, plasma is generated between two electrodes that are shaped as sheets electrically isolated from each other and arranged opposite to each other on the wall of the tank 120.

[0130] In some embodiments, the HV source may include a low voltage to HV transformer (not shown here) for generating the HV supplied to the electrode 172. In some embodiments, the HV transformer is a magnetic (inductive) step-up transformer as known in the art. In some embodiments, the HV transformer is a piezoelectric HV transformer, such as described in, for example, patent application publication No. WO 2021 / 144260. In some embodiments, the HV supplied to the electrode 172 is pulse modulated (square wave modulated) RF. Such pulse modulation is characterized by the duty cycle of the square wave, that is, the ratio of the wave "on" time to the total time period of the wave. By controlling the duty cycle of the modulation, employing such modulated RF HV allows relatively simple control of the power supplied to the electrode by the HV power supply, and therefore relatively simple control of the average power of the plasma produced.

[0131] Electrode 172 is preferably located inside inlet tube 160 with its tip 176 proximate inlet tube distal end 162. In some embodiments, tip 176 is retracted toward the interior of the tube a small distance, such as between 0.1 mm and 10 mm, including each value within the specified range.

[0132] During operation, the tank 120 is positioned in the tank 132 and the door 140 is closed, thereby sealing the interior of the tank 120 and preventing gas from escaping from the tank to the outside. The compressor 152 circulates air through the circulation system 150 by compressing the air into the inlet pipe 160 and forcing the gas toward the tank. When the gas is released from the inlet pipe distal end 162 into the liquid medium 110, bubbles 180 are formed and released into the top portion of the tank 120. The outlet pipe 154 collects the gas from the top portion of the tank and directs the gas back to the compressor 152.

[0133] During operation, HV power supply 170 is activated to generate HV, which is supplied to electrode 172 via wire 174. The HV at tip 176 applies a plasma-generating EM field between the tip and the surface of the liquid medium at distal end 162 of the inlet tube. The flow of air through the liquid via bubbles increases the liquid's surface area exposed to the plasma, increasing the activation rate of the liquid. Furthermore, the constant flow of air and the formation of bubbles agitate the liquid, exposing different portions of the liquid to the plasma over time. This prevents the risk of heating the liquid medium to undesirable levels.

[0134] In some embodiments, it may be advantageous to actively remove heat from the tank during plasma activation by cooling. In some such embodiments, the circulation system 150 may include a cooling module 180 for cooling the gas circulated by the circulation system. Such a cooling module may include a radiator and / or a heat exchanger and / or a compressor (none shown in the figures). In some embodiments, cooling the circulating gas may be performed directly, rather than through heat exchange with a coolant fluid, such as by using a radiator—by compressing the circulating gas and releasing the compressed gas through an orifice.

[0135] In some embodiments, the circulation system 150 is not a closed system. In an open circulation system (not shown here), the outlet port of the tank 120 can release the excited gas into the environment after the gas has passed through the liquid medium in the tank 120. In addition, in an open circulation system, the compressor 152 can collect air from the environment rather than from an outlet pipe fluidly connected to the tank.

[0136] In such embodiments employing an open circulation system, cooling the circulating gas can be performed directly by compressing the gas through a vortex tube. It should be noted that during operation, the vortex tube releases an output heat flow simultaneously with the output cold flow, which output heat flow can preferably be released to the environment. Therefore, using a vortex tube for cooling in an open circulation system is simpler.

[0137] In some embodiments described herein, where plasma-excited gaseous residue is released into the environment, an active filter can be employed to capture hazardous materials and prevent such materials from being released into the atmosphere. The active filter can be positioned upstream of the location where the gas is released into the environment—for example, downstream of the hot exhaust of a vortex tube—so that all gas passes through the active filter before being released into the atmosphere.

[0138] In some embodiments, gelation due to heating during plasma activation can be prevented by cooling the liquid medium. However, in some cases, this approach may not be as effective as cooling the circulating gas. Cooling the circulating gas removes heat from the source, i.e., from the area where the plasma is generated, and thus may be more efficient in some cases.

[0139] Figure 1E Schematically depicted are embodiments of an inlet tube 190 and a hollow needle 192 that is used as an electrode and configured to interface with an HV power source (e.g., Figure 1D The hollow needle 192 can be implemented, for example, by a syringe needle. The hollow needle 192 is sealed within the inlet tube 190 by a needle seal 194. As explained above, when air is compressed into the inlet tube 190, the compressed air is forced into the hollow needle 192 to be ejected from the needle distal end 196. Figure 1D and Figure 1E In an embodiment of the present invention, the plasma-generating EM field is applied directly to a gas stream flowing over a liquid medium.

[0140] Figure 2A Another embodiment of an apparatus 200 for plasma activation of a liquid medium 110 in a tank 210 is depicted. Apparatus 200 includes a housing 130 having a tank 132 and a door 140 configured to close over the tank. Apparatus 200 also includes an HV power supply 170 electrically connected to electrodes 172 via electrical wires 174. In some embodiments, apparatus 200 may lack a gas circulation system. Apparatus 200 differs from apparatus 100 in that it utilizes stirring to mix the liquid composition during plasma activation, thereby preventing heating and temperature increases in a single region of the liquid composition, thereby preventing gelation of the liquid composition during plasma activation. Stirring also promotes uniform activation of the liquid. Therefore, apparatus 200 includes a magnetic stirrer 220 configured to generate a rotating magnetic field within tank 132. Tank 210 includes a magnetic plate 222 adapted to react to the magnetic field. The magnetic plate 222 may be made of, for example, a ferromagnetic material such as iron, a permanent magnet, and may be coated with an inert material such as plastic to prevent oxidation or other chemical interactions with the liquid composition.

[0141] To prepare for use, the user removes the cover of the tank 210, which covers the tank during storage and transport. The tank can be inserted into the slot 132 and the door 140 can be closed. Closing the door allows the electrode 172 to be introduced into the tank 210, with the tip 176 advantageously positioned above the liquid surface. When the HV power supply 170 is activated, a plasma-generating EM field is applied between the tip 176 and the liquid surface, generating plasma in the region between them. During the plasma activation period, the magnetic stirrer 220 can also be activated, thereby rotating the magnetic plate 222 and stirring the liquid medium 110.

[0142] It should be noted that in some embodiments, the maximum power that can be used by a single electrode may be limited by the local temperature increase near the electrode. Attempts to increase the power beyond this limit may result in a local temperature increase of the liquid medium and, for example, gelation of a liquid composition capable of undergoing a liquid-gel phase transition, which may be undesirable. Therefore, in some embodiments, an apparatus for plasma activation of a liquid medium may include two electrodes instead of one. Such an apparatus with two electrodes may have the advantage of allowing the use of higher powers for plasma activation compared to an apparatus with a single electrode, thereby achieving a given activation level or a given RONS concentration in a shorter time.

[0143] For example, a device such as device 200 may include two electrodes, such as electrode 172, mutatis mutandis. In some such embodiments, the device may include two HV power supplies, such that each electrode may be electrically connected to a different HV power supply. Thus, each of the two electrodes may generate a plasma-activating EM field independently of the other electrode. In some embodiments, both electrodes may be electrically connected to a single HV power supply. In some such embodiments, HV may be supplied to both electrodes simultaneously, while in some embodiments, an HV distributor may distribute HV to the electrodes sequentially.

[0144] Advantageously, the electrodes may be arranged such that the distal end of each electrode (e.g. Figure 2A The distance between the tip 176 in the electrode 170 and the liquid surface is shorter than the distance between the electrodes and shorter than the distance between each electrode and the wall of the tank.

[0145] In some embodiments, the device 200 further includes a refrigerator 230 configured to remove heat from the tank 210 and the liquid medium therein. The refrigerator 230 can be thermally connected to the tank 210 via a heat exchanger 232. In some embodiments, the refrigerator 230 can be activated during the plasma activation period to prevent or reduce the temperature increase of the liquid composition and prevent gelation. In some embodiments, vapor compression cooling can be used. The advantage of conventional vapor compression cooling in a closed cycle including a compressor and an evaporator is relatively high efficiency; possible disadvantages are large size and a relatively long time to reach the target temperature. In some embodiments, a thermoelectric cooler (TEC) can be used. A thermoelectric cooler using, for example, Peltier cooling can have the advantages of relatively small size, lack of moving parts, and simple adaptation to the desired shape of the object to be cooled (e.g., tank 210). In some embodiments, a passive refrigerator can be used, as described in further detail below.

[0146] It should be noted that a refrigerator (such as refrigerator 230) and / or a stirrer (such as stirrer 220) (and corresponding magnetic plate 222) may be used mutatis mutandis with any of the embodiments described herein, e.g. Figure 1A-Figure 1D 、 Figure 2B wait.

[0147] Figure 2B Yet another embodiment of an apparatus 250 for plasma activation of a liquid medium 110 in a tank 260 is schematically depicted. Apparatus 250 employs vigorous shaking of the tank 260 during plasma activation to agitate and mix the liquid medium during plasma activation. In some embodiments, such vigorous shaking can generate a spray 262 within the tank. In some embodiments, the liquid medium can be partially or completely converted to foam.

[0148] The device 250 includes a housing 252 having a slot 254 and a door 270 configured to close over the slot. The device 250 also includes an HV power supply 170 electrically connected to a sliding pad 272 via an electrical wire 174. The tank 260 includes a removable tank cover 264, which includes a sliding contact 276 on the outer surface of the tank cover. The sliding contact 276 is electrically connected to an electrode 280, which is attached to the inner surface of the tank cover and has a tip 282. In some embodiments, the sliding contact 276 is electrically connected to the electrode 280 via a feedthrough (not shown here) in the tank cover 264. In some embodiments, the tank cover 264 is made of a dielectric material. In some embodiments, the tank cover 264 is metal and the electrode 280 is electrically isolated from the tank cover. In some embodiments, the tank cover 264 is metal and is electrically connected to the electrode.

[0149] The device 250 also includes a vibration generator 290 mechanically connected to a tank holder 292. For use, the tank 260 can be positioned in the slot 254 and attached to the tank holder 292. When the door 270 is closed, the sliding contact 276 contacts the sliding pad 272, thereby electrically connecting the electrode 280 to the HV power supply 170. During plasma activation, the vibration generator 290 is also activated to vigorously vibrate or violently shake the tank 260. Activation of the power supply 170 generates plasma over a relatively large area around the tip 282. This is likely due to the formation of a spray 262 in the space around the tip, which creates a large number of discharge tracks between the electrode and the liquid composition. The interface between the plasma and the large number of droplets in the spray significantly increases the surface area of ​​the liquid composition in contact with the gaseous plasma and accelerates the activation of the liquid. If the liquid medium is the liquid composition of the present invention, the mixing of the liquid caused by the shaking of the tank 260 during plasma activation further prevents heating and temperature increases in one area of ​​the liquid composition, thereby preventing gelation. After plasma activation, the user can remove the can from the can holder 292, remove the can cover 264, and use the liquid medium as desired.

[0150] In some embodiments, additionally or alternatively, the vibration generator 290 can apply a rotational displacement to the tank 260. In some such embodiments, such rotational displacement can be back and forth. In some such embodiments, the tank 260 can include fins or blades (not shown here) to help mix the liquid medium during such rotational displacement.

[0151] In some embodiments, the apparatus 250 may also include a cooler or refrigerator (not shown here) for cooling the liquid composition during plasma activation. In some embodiments, such a cooler may be thermally connected to the tank 264 via the tank holder 292 or via a dedicated heat exchanger in thermal contact with the tank.

[0152] Example 4

[0153] Transportable sealed canister containing a liquid composition

[0154] Figure 3A An embodiment of a sealed canister 300 containing a liquid medium 110 according to the teachings herein is schematically illustrated. The sealed canister 300 is configured for storage and / or transport, and for use in an apparatus for plasma activation of a liquid medium according to the teachings herein. In some embodiments, the sealed canister 300 is substantially made of a dielectric material such as glass or plastic. In some embodiments, the sealed canister 300 is made of metal. In some such embodiments, the canister wall can be electrically connected to ground potential during plasma activation.

[0155] The sealed can 300 is configured to be hermetically sealed during storage or transportation so that the liquid medium 110 cannot escape from the sealed can and cannot be contaminated by pollutants that enter the sealed can from the outside. In some embodiments, the sealed can 300 may include a cover 312 for the opening of the sealed can. In some embodiments, the cover 312 can be loosened by the user to connect the interior of the sealed can 300 with the ambient fluid. In some embodiments, the cover 312 can be completely removed from the opening of the can.

[0156] The sealed canister 300 includes a hollow needle 320, such as a syringe needle. The hollow needle 320 is electrically conductive, typically made of metal, and in some embodiments has a pointed distal end 322. The sealed canister 300 also includes a gas port 330 in fluid communication with the lumen of the hollow needle. Thus, the gas port 330 enables gas to flow into the canister through the hollow needle 320. The gas port 330 is sealable and is therefore configured to be sealed when the sealed canister is in storage or during transport, and can be opened by the user for gas flow, for example, during plasma activation as described below.

[0157] The sealed canister 300 further includes an HV connector 340 electrically connected to the hollow needle 320. The HV connector 340 is configured to electrically connect a high voltage to the hollow needle. Upon receiving HV, such as from an HV power source, the hollow needle can apply an EM field within the sealed canister, preferably between the distal end 322 and the surface of the liquid medium 110, to generate a plasma. In some embodiments, the plasma is generated by the hollow needle in an arc mode.

[0158] In some embodiments, the gas port and HV connector may be embodied in a single connector.

[0159] In some embodiments, the sealed canister 300 includes an exhaust port 350 that is configured to enable gas to flow from the sealed canister to the environment. The exhaust port 350 can be sealed during storage and / or transportation of the sealed canister 300 and can be opened by the user for gas flow, for example, during plasma activation as described below. In some embodiments, the exhaust port 350 can be sealed by a foil that can be removed or torn to open the port. In some embodiments, the exhaust port 350 can be configured to be fluidically connected to a gas flow channel (not shown here) (e.g., a tube) to direct the outflowing gas to further processing, for example, for filtering before releasing the gas into the environment, or for recycling, as described above Figure 1DIn some embodiments, the exhaust port 350 can be configured to release the outflowing gas into the environment without further treatment. In some embodiments, the outflowing gas can be released into the environment via the opening of the sealed can (for example, by loosening the cover 312).

[0160] In some embodiments, the sealed jar 300 includes a magnetic plate 222 that allows the liquid composition to be stirred by a magnetic stirrer, as described above in Figure 2A Additionally or alternatively, the sealed tank may include fins or blades (not shown here) to help mix the liquid medium during shaking or rotating the tank, as described above in Figure 2B Described in .

[0161] The sealed can 300 can be used in conjunction with an apparatus 360 for plasma activation of a liquid medium, such as Figure 3B The device 360 ​​may include a housing 370 and a slot 372 in the housing, the slot 372 being configured to receive a device as described above for example Figure 2A The described sealing pot. The door 374 can be used to lock the sealing pot 300 in the slot.

[0162] The sealed canister can be preferably manufactured and suitably stored under aseptic conditions. When, for example, a clinic needs, the sealed canister can be transported to the clinic. In the clinic, the sealed canister can be placed in the groove 372 of the device for plasma activation. The gas port 330 can be connected to an airflow source, such as a compressor 380, via an inlet channel 382. The HV connector 340 can be electrically connected to the HV power supply 170. The outflow of gas from the canister can be achieved, for example, by opening an exhaust port 350 or, for example, by loosening the cover 312 of the sealed canister. In some embodiments, the exhaust port 350 can be fluidically connected to the outlet channel of the device, which can include filters (both not shown here) for absorbing or removing the harmful residues of plasma before the gas is released into the environment.

[0163] Plasma activation of the liquid composition can then be achieved by supplying HV to the hollow needle 320, preferably while simultaneously flowing gas through the hollow needle into the sealed canister, and while simultaneously agitating the liquid composition, for example, by stirring the liquid using the device's magnetic plate 222 and magnetic stirrer 220, or by any other method. An active refrigerator 390 can be used to maintain the liquid composition at a sufficiently low temperature using a heat exchanger 392. According to some embodiments, a passive refrigerator can be used as further described below.

[0164] After plasma activation is complete, the sealed canister can optionally be removed from tank 372. The sealed canister can be opened, for example, by removing cover 312 from the canister's opening, thereby exposing the activated liquid medium therein. The activated liquid medium can then be obtained for application, for example, by drawing the liquid into a syringe and then applying the liquid to the desired area of ​​the body.

[0165] Example 5

[0166] Using a Nozzle

[0167] According to aspects of the present invention, nozzle can be used for the liquid composition of activation of the present invention is applied on body parts.In some embodiments, liquid composition can be poured into spray bottle, and then spray bottle can be used for applying material.In some embodiments, nozzle head (for example, such as the nozzle head of spray bottle) can be assembled on the tank for plasma activated liquid.In some embodiments, nozzle head can include positive displacement pump (positive displacement pump) (possibly but not necessarily manual), for dispensing material by nozzle, thereby produces spray.For example, as described above, sealed tank 300 can be used for plasma activated liquid composition wherein, cover 312 can be removed subsequently and nozzle head can be attached to tank, for example, be screwed on tank, for liquid composition is applied on body parts.

[0168] Example 6

[0169] Foaming liquid compositions

[0170] In some embodiments, foaming the liquid composition of the present invention can convert the liquid into a relatively thick foam. "Relatively thick" herein means that the foam is at least as thick as a gel according to the teachings herein.

[0171] In some embodiments, such foaming of the liquid may require vigorous agitation of the liquid composition—e.g., rapid stirring—or a combination of vigorous agitation and strong air flow onto the liquid composition (e.g., from a hollow needle). The terms "vigorous agitation" and "strong air flow" are used herein in a relative sense, meaning that the agitation or flow intensity must be above a respective threshold to produce foam. Thus, according to aspects of the present invention, there is provided a plasma-activated foam that can be produced by plasma-activating a liquid composition while vigorously agitating the liquid and, possibly, simultaneously causing a strong air flow to flow onto the liquid.

[0172] The inventors have discovered that an exemplary liquid composition of the present invention prepared by mixing 20.00% (wt.) poloxamer 407 with 5.00% poloxamer 188 and 75% water in a total volume of 10 ml is converted into a foam after mixing using a magnetic stirrer at a speed greater than 1000 RPM for 3 minutes while maintaining a temperature below 5° C. The foam is stable at a temperature below 5° C. for about 10 minutes, after which it returns to a liquid form.

[0173] It should be noted that plasma-activated foams can be very useful because the liquid-foam transition hardens the material. In other words, the foam produced can be viscous enough to allow it to be easily applied to a body part (e.g., a portion of the skin) and to allow stability of the material at the application site without the risk of foam migration or flow. In some embodiments, regardless of the material's ability to undergo a liquid-gel phase transition, this hardening of the material due to foaming is possible. In such embodiments, the liquid composition used for plasma activation should not necessarily be able to undergo a liquid-gel phase transition, and thus the liquid composition can be simpler to manufacture and / or cheaper (compared to a liquid composition that can undergo such a phase transition). However, it should be understood that even when used as a foam, a liquid composition that undergoes a liquid-gel phase transition according to the teachings herein may also be advantageous due to its stability over a long period of time (e.g., several days).

[0174] Example 7

[0175] Passive cooler

[0176] Figure 4A and Figure 4B An embodiment of a passive refrigerator is schematically illustrated which may be used with the apparatus for plasma activation of a liquid medium as described above according to aspects of the invention.

[0177] Figure 4AAn embodiment of a passive cooler 400 is illustrated, which includes an ice pack 410 in the form of a torus surrounding a can 420. In some embodiments, the can 420 can be open at the top, as depicted; in some embodiments, the can 420 can be closable by a cover (not shown here). In some embodiments, the can 420 is detachable from the ice pack 410. In such an embodiment, the ice pack has a cooler groove 422 that is sized and configured to accommodate the can when the can is inserted into the groove. Advantageously, the cooler groove 422 is sized and configured to establish good thermal contact between the ice pack and the can. In some embodiments, when the can 420 is inserted into the groove, a flexible metal member (not shown here) within the groove 422 adapts to the outer shape of the can 420 so as to establish good thermal conduction between the ice pack and the can. In some embodiments, the ice pack and the can are attached together, with the wall of the can also serving as the wall of the ice pack.

[0178] The embodiment of the ice pack 410 can have a variety of shapes and forms, as will be appreciated by those skilled in the art. The outer shape of the ice pack 410 can be as follows: Figure 4A In some embodiments, the ice pack 410 may also extend in the area below the can 420 so as to extract heat from the can 420 not only through the side walls of the can but also from the bottom wall of the can (the bottom of the can).

[0179] The ice pack 410 comprises a coolant material having a melting temperature of about zero degrees Celsius, for example between about -10°C and about +5°C. Advantageously, but not necessarily, the material may have a relatively high latent heat of fusion (herein referred to as 'latent heat'), for example, greater than 200 J / gram or greater than 250 J / gram or even greater than 300 J / gram. For example, the ice pack 410 may be filled with water, which has a freezing temperature of about 0°C and a latent heat of about 333 J / gram. Additionally or alternatively, the ice pack 410 may be filled with a gel material such as is typically found in commercial ice packs for home use, such gel being obtained, for example, by adding additives such as hydroxyethyl cellulose, sodium polyacrylate or vinyl-coated silica gel to water.

[0180] Ice pack 410 can be made of a rigid material or a semi-rigid material. Optionally, ice pack 410 can be made substantially of plastic, such as commercial ice packs for home use. Canister 420 can be made of a hard dielectric material or a conductive material such as a metal.

[0181] Figure 4B4 is an exploded view of an embodiment of a passive cooler 430. Cooler 430 includes an ice pack 440 surrounding a cooler tank 442, and a removable canister 450 sized and configured to be inserted into cooler tank 442. Ice pack 440 differs from ice pack 410 of cooler 400 in having an outer chamber 444 for each of the two different materials and an inner chamber 446 surrounded by the outer chamber. Advantageously, outer chamber 444 stores a coolant material having a freezing temperature of approximately zero degrees Celsius and preferably a high latent heat, as described above for cooler 410. Inner chamber 446 stores a material that remains in a liquid phase at a temperature below the freezing temperature of the coolant in outer chamber 444. In some embodiments, the material in inner chamber 446 can remain liquid at temperatures below -10°C, and even more preferably below -24°C. Advantageously, the liquid in inner chamber 446 has a relatively high thermal conductivity. For example, the liquid can be distilled water with an antifreeze additive such as ethylene glycol or propylene glycol, or the like.

[0182] The inner chamber 446 can advantageously be made of a soft or flexible material. Thus, when the can 450 is inserted into the groove 442, the liquid contents of the inner chamber together with the softness of its walls ensure that the groove adjusts to the outer surface of the can, thereby ensuring good thermal contact between the interior and the can.

[0183] Figure 4C The apparatus 460 for plasma activation of a liquid medium is illustrated, incorporating a passive cooler 470 as described herein, positioned in a slot 462 of a housing 464 of the apparatus. A removable canister 480 is shown in the cooler slot 472 in thermal contact with a cold portion 474 (e.g., an ice pack) of the passive cooler. The apparatus 460 also includes an electrode 172 electrically connected to an HV power source 170 and configured to apply a plasma-generating EM field between the electrode and the surface of the liquid medium 110. The apparatus also includes a magnetic stirrer 220 as described above.

[0184] Before the device is operated, the ice pack of the passive refrigerator can be advantageously cooled to below the freezing temperature of the coolant. When the coolant is frozen, the ice pack can be placed in the tank 462 and the canister 480 can be positioned in the refrigerator tank 472.

[0185] According to an example, 50cc of a liquid composition having an initial temperature of approximately 0°C is plasma treated in apparatus 460. The power flowing into the liquid composition by the electric field generating the plasma, by stirring, and by heat diffusion from the environment totals approximately 100W. An ice pack containing approximately 300cc of a coolant having a latent heat of approximately 330J / gram can absorb the relevant heat from the liquid composition for a duration exceeding 15 minutes before completely melting. Heat transfer from the liquid composition to the coolant occurs across the interface surfaces (i.e., the tank wall and any ice pack wall). A total interface surface of approximately 150cm^2 and a total heat conduction of approximately 50W / °C*m result in a temperature gradient across the interface surfaces of less than 5°C.

[0186] After the plasma treatment is complete, the activated liquid medium can be removed for use, and the ice pack can be removed to a refrigerator to chill the coolant.

[0187] Example 8

[0188] Treating skin disorders

[0189] PAMs prepared by any of the methods described above can be used to treat skin disorders other than otitis in pets and other domestic animals. Such skin disorders can be caused by yeasts or fungi such as Malassezia (causing Malassezia dermatitis) and ringworm or by bacteria (e.g., Staphylococcus intermedius pyoderma or Alabama rot caused by E. coli toxins). Skin disorders can also be caused by parasites, such as sarcoptic mange (scabies), mange caused by Demodex mites (Demodicosis) and Cheyletiella. Viral skin disorders, such as warts, can also be treated according to the teachings herein. According to some embodiments, the PAMs of the present invention can be used to treat skin disorders due to autoimmune system dysfunction in combination with external or environmental triggers, such as dermatitis, such as atopic dermatitis. According to some embodiments, the PAMs of the present invention can be used to treat skin disorders in humans.

[0190] In order to use PAM on naked skin according to the teachings of this paper, a liquid composition with similar viscosity or higher viscosity (compared to the liquid composition prepared for treating ear infection as described above) can be prepared and activated by plasma. The liquid composition can then be applied directly to the skin or used to soak a gauze or bandage applied to the skin. In some embodiments, the liquid composition can be applied to the skin by spraying as described above. In some embodiments, the liquid composition can be foamed and then applied to the skin. In such cases, the hardness of the foam can help ensure the stability of the activated composition on the skin site until the material is converted into a gel.

[0191] In some embodiments, in response to reaching an elevated temperature, such as body temperature, the liquid composition can gel as described above. In some embodiments, gelation can be achieved by mixing the activated liquid composition with a crosslinking agent. Additionally or alternatively, crosslinking can be achieved by exposing the PAM to visible light or UV light (see, e.g., Lim K.S. et al., "Visible Light Cross-Linking of Gelatin Hydrogels Offers an Enhanced Cell Microenvironment with Improved Light Penetration Depth," Macromolecular Bioscience, Vol. 19, No. 6, 1900098, https: / / doi.org / 10.1002 / mabi.201900098 ).

[0192] Example 9

[0193] In vitro reduction of bacterial loads in samples taken from infected ears

[0194] The compositions as outlined in Table 1 (Example 2, Figure 2A The plasma activated composition was applied to secretions from infected ears of dogs incubated on agar plates. Figure 5 , right panel) compared to when using activated gel ( Figure 5 , left panel), up to a 3.5-log reduction in bacterial load was observed.

[0195] Example 10

[0196] Pam in the treatment of periodontal infection

[0197] Periodontal disease is a common chronic inflammatory condition worldwide. Often, poor or ineffective oral hygiene leads to gingivitis, an inflammation characterized by redness and swelling of the gums. In some cases, gingivitis may progress to periodontitis, a bacterial infection that stimulates a host response, leading to loss of the tooth's supporting structures (soft and hard tissues). As gingival fibers are destroyed, the gum tissue separates from the teeth, resulting in periodontal pockets. If left untreated, periodontitis is one of the leading causes of tooth loss in adults. It is a difficult disease to treat and requires specialized mechanical subgingival cleaning, known as scaling and root planing (SRP). If non-surgical treatment is unsuccessful, surgical intervention may be necessary. SRP is often accompanied by local antimicrobial therapy at the site of infection or systemic antibiotics. An example of a non-antibiotic-based local antimicrobial treatment is provided by a slow-release chlorhexidine chip, which is positioned in the periodontal pocket by the treating dentist (see, for example, https: / / www.rxlist.com / periochip-drug.htm#description and https: / / www.periochip.com). The chip releases chlorhexidine in a biphasic manner, releasing approximately 40% of the chlorhexidine within the first 24 hours and then releasing the remaining chlorhexidine approximately linearly over 7-10 days.

[0198] Subgingival destructive inflammation can sometimes develop in the soft tissue surrounding a dental implant (peri-implantitis). As a result, the alveolar bone (hard tissue) surrounding the implant for retention purposes can be lost over time. Therefore, peri-implantitis and periodontitis can be treated according to the teachings herein, which, for simplicity, will relate to periodontal pockets.

[0199] According to some embodiments of the present invention, a plasma-activated medium (PAM) can be applied to the periodontal pocket of a subject in need thereof. While in the periodontal pocket, the PAM gradually releases RONS, promoting disinfection around the pocket. The PAM is biodegradable, and after a predetermined period of time (typically several days), the PAM decomposes and its residues naturally wash out of the pocket.

[0200] In some embodiments, the PAM can be a composition comprising a synthetic biocompatible polymer capable of in situ gelation. A preferred stimulus for gelation can be an increase in temperature from typical room temperature (e.g., about 26° C.) to typical body temperature (e.g., about 37° C.). The liquid composition can be activated by plasma as described in Example 2 above. The resulting PAM can then be applied to the periodontal pocket, where the composition is converted into a gel and stabilized in situ. According to some embodiments, the liquid composition substantially fills the periodontal pocket, effectively preventing food debris and other potential contaminants from entering the pocket.

[0201] Due to the moist environment of the periodontal pocket, appropriate measures should be taken to prevent dissolution of the resulting gel.In some embodiments, higher concentrations of poloxamer (compared to the concentration required for gelation in a dry environment) may be sufficient to provide a suitably stable gel.

[0202] In some embodiments, a further hardening mechanism can use optical crosslinking. Optical crosslinking can use UV light or visible light. In such embodiments, the liquid composition can be injected into the periodontal pocket and then crosslinked by exposure to light. The light can be applied via an optical fiber gently inserted into the periodontal pocket and scanned over the top surface of the filling. In some embodiments, in response to the elevated temperature as explained above, the liquid composition can be converted into a gel having a first hardness or viscosity level, and the gel can then be further hardened or cured to a higher viscosity level by light-induced crosslinking.

[0203] In some embodiments, the liquid composition can be plasma activated as described above and then cured in a mold outside the periodontal pocket. The resulting molded part, which may be in the form of a thin sheet, can be inserted into the periodontal pocket and secured therein. Curing the PAM outside the periodontal pocket has the advantage of employing a curing and hardening process that can achieve the desired hardness level, but which may not be used when the PAM is inside the patient's mouth, such as chemical cross-linking. However, it may have the disadvantage of only producing a PAM of a predetermined shape and size, i.e., it may not completely fill the periodontal pocket.

[0204] Example 11

[0205] Treatment of premalignant tumors and malignant tumors

[0206] Pancreatic cancer is the third most common cause of cancer death in the United States. Because the pancreas is located deep in the body, behind the stomach, it is not easily palpated, and tumors may often be detected at a relatively late stage, after the tumor growth has already affected the normal function of the pancreas itself or other nearby organs. Even when tumors are detected at a relatively early stage, one- and five-year survival rates are relatively low because surgery is currently the only medical procedure that may result in a cure, and because, due to the pancreas' location and other factors, surgery is complex and not always possible.

[0207] A small percentage of pancreatic cancers develop from cysts, although the majority of pancreatic cysts do not become cancerous. This situation creates a dilemma regarding how to treat a pancreatic cyst once it has been detected. Typically, the sequence of actions involves determining the type of cyst to determine whether it is likely to be cancerous and, in suspected cases, performing a biopsy. If the biopsy reveals a noncancerous or precancerous cyst, the recommended strategy is usually watchful waiting, attempting to avoid complex surgery to remove the cyst, as cysts generally do not become cancerous over a short period of time even without treatment. However, it is clear that such a watchful waiting period necessarily carries both burdens and risks.

[0208] Thus, according to aspects of the present invention, a method for treating pancreatic cysts using PAM is provided in accordance with the teachings herein. The method is intended to reduce or eliminate inflammation in and around the cyst, thereby eliminating the cyst itself. Additionally or alternatively, the method is intended to selectively reduce the number of precancerous cells in the cyst or even eliminate precancerous cells. The method comprises delivering a plasma-activated composition according to the teachings herein to the cyst (or lesion or tumor). The plasma-activated composition is in liquid form during delivery and gels after deployment in the patient.

[0209] Figure 6 An embodiment of an endoscope 600 for delivering a plasma-activated liquid composition to a cyst 610 in a patient's pancreas 612 is schematically depicted. The endoscope is advanced through the patient's mouth and esophagus into the patient's stomach 618 (depicted here in a cross-sectional view). The endoscope can conveniently include an ultrasound transceiver 620 at its distal end 622, which is functionally associated with an ultrasound device including a screen positioned outside the patient's body and configured to provide an ultrasound image of the area surrounding the transceiver 620 to a medical practitioner. The endoscope also includes a working channel configured to extend through the lumen of the endoscope. As is known in the art, the working channel enables a practitioner to advance a working tool through the working channel and through an exit opening 630 near the distal end 622 and manipulate the working tool using a control unit outside the patient's body.

[0210] To deliver the plasma-activated composition to the cyst 610, the practitioner can use an ultrasonic transceiver 620 to search for and detect the cyst 610 and position the distal end 622 in the appropriate position relative to the cyst for injection. The practitioner can then advance the injection needle 640 through the working channel and the working channel's outlet opening 630. The injection needle is in fluid communication with a syringe (not shown here) located outside the patient's body via an elongated tube passing through the working channel. Prior to the procedure, the liquid composition can be plasma-activated, for example, as described in Examples 2-7 above, and the syringe can be filled with the plasma-activated composition in liquid form. By manipulating the injection needle, the practitioner can penetrate the pancreatic cyst through the stomach wall. Furthermore, using a syringe, the practitioner can inject the plasma-activated composition into the cyst through the tube and injection needle.

[0211] According to some embodiments, the working channel can be continuously washed with cold saline during surgery to prevent premature gelation of the liquid composition in the tube within the working channel. The saline can be cooled to about 5°C and can flow through the working channel to drain into the stomach through the outlet opening 630.

[0212] According to some embodiments, the cyst may be drained, for example, by injection needle 640, prior to injecting the composition into the cyst.

[0213] Example 12

[0214] Treatment of late-stage malignant tumors—tumor resection in the bladder

[0215] Following resection of a tumor in the bladder or colon, a plasma-activated composition according to certain embodiments of the present invention is applied to the resection site to prevent tumor recurrence.

[0216] Transurethral resection of bladder tumor (TURBT) is the gold standard for surgical treatment of bladder cancer. Unfortunately, the postoperative recurrence rate of bladder cancer is high. The current standard of care to further reduce the recurrence of bladder cancer is intravesical chemotherapy (ICT). An alternative that has been proposed is to flush the bladder with saline or water (see A. Mahran et al., "Bladder irrigation after transurethral resection of superficial bladder cancer: a systematic review of the literature" Can. J. Urol. 25 (6), December 2018, p. 9579).

[0217] According to aspects of the present invention, plasma-activated compositions according to the teachings herein can be used for postoperative treatment to reduce or eliminate the possibility of cancer recurrence. According to an embodiment, an aqueous solution (e.g., saline) is plasma-activated by exposing the solution to a gaseous plasma. According to some embodiments, the aqueous solution is exposed to a plasma generated in air at room pressure. According to some embodiments, the gaseous plasma is generated in a dielectric barrier discharge (DBD) mode by applying an electromagnetic field between an anode located above the surface of the solution and a cathode located below the solution or electrically coupled to the solution. According to some embodiments, the anode is coated with an electrically insulating layer to provide a DBD mode for generating plasma. In some embodiments, the line of sight between the anode and the solution surface is interrupted by a dielectric layer between the anode and the solution surface.

[0218] According to some embodiments, the aqueous solution is plasma-activated by a plasma jet. To this end, a plasma generator can be positioned above the surface of the solution, and an airflow can be manipulated through the plasma generation region of the plasma generator to be excited by the plasma. The excited airflow is then focused to form a jet directed toward the surface of the solution. According to some embodiments, the airflow comprises an inert gas such as helium or argon or a gaseous mixture thereof.

[0219] According to some embodiments, the solution may be stirred during exposure to the gaseous plasma to accelerate dissolution of plasma excited products in the solution.

[0220] After the aqueous solution is plasma activated, for example, as described above, the plasma-activated solution can be used to irrigate the bladder. In some embodiments, bladder irrigation is performed using a catheter passed through the urethra, as is known in the art. According to some embodiments, the plasma-activated solution is maintained in the bladder for a period of time between 1 minute and 2 hours, including each value within the specified range.

[0221] Example 13

[0222] Treatment of late-stage malignancy – polypectomy in the colon

[0223] Rectal villous adenoma is a type of colon polyp (of the five commonly considered types) that accounts for approximately 15% of polyps detected in colon cancer screening tests and has a high risk of becoming cancerous. In cases of complete resection where the specimen is found to be cancerous on pathology, approximately 40% of cases experience recurrence (see, e.g., Sungeyun David Cho, "Treatment Strategies and Outcomes for Rectal Villous Adenoma From a Single-Center Experience", Arch Surg., 143(9), 2008, p. 866).

[0224] According to aspects of the present invention, plasma-activated compositions according to the teachings herein can be used for post-operative treatment to reduce or eliminate the likelihood of cancer recurrence. Liquid compositions capable of gelling at or near body temperature are plasma-activated, for example as described above in Example 2. The method of applying the composition to the desired location is similar to that described above with respect to Figure 6 The method is described. A syringe fluidically connected to an elongated tube is filled with a composition. A colonoscope is manipulated through the colon until the distal end of the colonoscope is adjacent to the surgical field, and the elongated tube is advanced along the working channel of the colonoscope. The composition is then injected through the tube into the surgical field, where it gels on the colon wall. A stent can be positioned where the composition is applied to protect the gel from washing away from the colon wall.

[0225] Thus, according to aspects of the present invention, there is provided a system for providing a plasma activated medium (PAM) for medical use. Figure 1B The capsule 10 in the activation unit 60; Figure 1D 、 Figure 2A 、 Figure 2B 、 Figure 3B and Figure 4C The system includes a container (10, 120, 210, 260, 300, 480) configured to contain a liquid medium (12, 110). The system also includes a high voltage power supply ( Figure 1B 72 of them, Figure 1D 、 Figure 2A 、 Figure 2B 、 Figure 3B and Figure 4C 170) electrically connected to the electrode ( Figure 1B 38 of them, Figure 1D 、 Figure 2A and Figure 4C 172 of them, Figure 1E 192 of them, Figure 2B 280 of them, and Figure 3A and Figure 3B 320) in, the electrode is configured to apply an electromagnetic field that generates a plasma inside the container. The system further includes actuators (66, 152, 220, 290) that are configured to cause agitation of the liquid medium inside the container. And the system further additionally includes a refrigerator (230, 390, 400, 430, 470) having cold portions (232, 392, 410, 446, 474 respectively), the refrigerator being configured to be thermally coupled to the liquid medium inside the container. It should be noted that any refrigerator can be used with Figure 1B the system of. Refrigerators 230 and 390 can be used with Figure 2B the system of. Refrigerators 400 and 430 can be used with Figure 4C the system of. According to an aspect of the present invention, the system is configured to generate a plasma inside the container at atmospheric pressure while agitating the liquid medium and maintaining it at a temperature below room temperature.

[0226] According to some embodiments, the liquid medium is a liquid composition containing a biocompatible polymer, wherein the liquid medium is frozen at a temperature below T1 and assumes a gel form at a temperature above T2, where T1 < T2. According to some embodiments, T1 is between about -5 °C and about +20 °C, and T2 is between about +5 °C and about +45 °C. According to some embodiments, T1 is below about +15 °C, and T2 is above about +25 °C.

[0227] According to some embodiments, the biocompatible polymer is synthetic.

[0228] According to some embodiments, refrigerator 230 or 390 may include a vapor compression refrigerator. According to some embodiments, the refrigerator includes a vortex tube. According to some embodiments, refrigerator 230 or 390 includes a thermoelectric cooler. According to some embodiments, refrigerators 400, 430, and 470 include ice packs (410, 444, 474 respectively) containing a coolant. According to some embodiments, refrigerators 400, 430, and 470 are detachable from the housing.

[0229] According to some embodiments, the actuator ( Figure 1B 66 in Figure 1D 152 in Figure 2A , Figure 3B and Figure 4C 220 in Figure 2B and Figure 1B 72 in Figure 1D , Figure 2A , Figure 2B , Figure 3B and Figure 4C 170 in) and the HV power supply ( Figure 1B 62 of them, Figure 1D and Figure 2A 130 of them, Figure 2B 252 of them, Figure 3B 370 and Figure 4C According to some embodiments, the wall of the container is dielectric. According to some embodiments, the wall of the container is metallic. According to some embodiments, the wall of the container is electrically connected to ground potential.

[0230] According to some embodiments, the electrode is within the container. According to some embodiments, the electrode ( Figure 1B 38 of them, Figure 1D 、 Figure 2A and Figure 4C 172 of them, Figure 1E 192 of them, Figure 2B 280 of them, and Figure 3A and Figure 3B The system further comprises a second electrode electrically connected to a second high voltage power supply and configured to apply an electromagnetic field within the container that generates plasma.

[0231] According to some embodiments, the actuator is an agitator (220), and the agitation is achieved by stirring the liquid medium. According to some embodiments, the agitator is a magnetic agitator, and the container includes a magnetic plate (222). According to some embodiments, the actuator is a vibration actuator (290), which is configured to vibrate and / or rotate the container, thereby shaking the liquid medium. According to some embodiments, the actuator is a compressor (66, 152), which is configured to force air flow onto the liquid medium, thereby stirring the liquid medium. According to some embodiments, the gas is released in the liquid medium in the container, thereby generating bubbles in the liquid medium. In some embodiments, two or more methods can be used together to stir the liquid medium. For example, a compressor can be used to generate bubbles in the liquid while using an agitator to stir the liquid medium.

[0232] In some embodiments, the electrodes may be vibrated or displaced so that plasma is not generated at a single location in the liquid medium, thereby helping to reduce localized heat buildup in the liquid medium.

[0233] According to some embodiments, the system ( Figure 1B , device 100, device 360) further comprises a compressor connected to the container fluid via the container's inlet channel and gas inlet port (respectively 34, 162, 330), the compressor being configured to force gas into the container. According to some embodiments, for example, in Figure 1B and Figure 3B In some embodiments, the gas is air extracted from the environment. Figure 1D As shown, the compressor draws gas from the container via an outlet passage (154) fluidly connected to a gas outlet port (156) of the container, thereby circulating the gas through the container.

[0234] According to some embodiments, for example, Figure 1B 、 Figure 1D 、 Figure 1E and Figure 3B According to some embodiments, the electrodes (192, 320) are formed as open-ended tubes fluidly connected to the compressor, thereby generating the plasma-generating EM field along the gas flow between the distal end of the tube and the liquid medium.

[0235] According to some embodiments, the inlet passages (e.g., tank port 70, inlet passages 160, 382) can include filters. Such filters in the inlet passages can be used to ensure the sterility of the gas (e.g., air) forced into the container by filtering out biological contaminants such as viruses or bacteria. According to some embodiments, the outlet passages (e.g., outlet port 350) can include filters. Such filters can be used to absorb or filter out harmful remnants of the plasma, such as ozone, before releasing the gas into the environment.

[0236] According to aspects of the present invention, a transportable closed container 300 is also provided. The closed container includes a liquid medium stored in the container, intended for medical use, and an electrode (320) electrically connected to an HV connector (340). The electrode is configured to apply an EM field within the container that generates a plasma when receiving a high voltage via the HV connector. The container is bio-sealed to prevent bacteria or viruses from penetrating the container. According to some embodiments, the container is hermetically sealed.

[0237] According to some embodiments, the liquid medium is sterile. According to some embodiments, the liquid medium comprises a biocompatible polymer forming a liquid composition capable of undergoing a phase transition to a gel form upon stimulation.

[0238] According to some embodiments, the walls of the container are dielectric. According to some embodiments, the walls of the container are metallic. According to some embodiments, the container has an internal volume between 5 cc and 50 cc.

[0239] According to some embodiments, the container further comprises an inlet port (330) configured to enable the flow of gas into the container.

[0240] According to some embodiments, the electrode 320 comprises a metal tube in flow communication with the inlet port and having a distal portion within the container. According to some embodiments, the metal tube has a pointed tip at its distal end 322.

[0241] According to some embodiments, the container further comprises a magnetic plate (222) immersed in the liquid medium and configured to cause agitation of the liquid when subjected to a corresponding magnetic field.

[0242] According to some embodiments, the container further comprises an outlet port (350) allowing gas to flow out of the container. According to some embodiments, the container further comprises a removable cover (312), wherein the inlet port and the HV connector are assembled to the cover.

[0243] According to aspects of the present invention, a portable, passive cooler (400, 430, 470) configured to refrigerate a container is also provided. The passive cooler includes an ice pack (410, 444, 474, respectively) having at least one chamber for containing a coolant. The coolant preferably has a freezing temperature between -10°C and +5°C. The at least one chamber of the ice pack is shaped around a groove (422, 442, 472, respectively) in the ice pack, wherein the groove is configured and sized to receive the container therein.

[0244] According to some embodiments, the passive cooler (e.g., cooler 400) further comprises a container (420) for the ice pack attached to the tank, wherein the container is advantageously metallic and in direct contact with the coolant. According to some embodiments, the container 420 comprises an openable cover for the container. According to some embodiments, the container 420 further comprises an HV connector and an electrode electrically connected to the HV connector, such as in Figure 3A The electrodes are preferably electrically insulated from the metal container and are configured to apply a plasma-generating EM field in the container when receiving a high voltage from an HV power source.

[0245] According to some embodiments, the groove of the passive refrigerator is a through hole in the ice pack. According to some embodiments, the groove is formed as a depression in the ice pack. According to some embodiments, the passive refrigerator (430) further includes a non-rigid member surrounding the groove, the non-rigid member being configured to adjustably contact the container when the container is placed in the groove, thereby establishing thermal contact between the ice pack and the container. According to some embodiments, the non-rigid member includes a flexible metal portion. According to some embodiments, the non-rigid member includes at least one second chamber 446, at least one wall of which is non-rigid. The at least one second chamber preferably contains a material that is liquid at the freezing temperature of the coolant. According to some embodiments, at least one non-rigid wall is soft.

[0246] According to aspects of the present invention, there is also provided a liquid composition comprising a biocompatible polymer, wherein the liquid composition undergoes a phase transition to a gel form upon stimulation. The liquid composition further comprises reactive substances H2O2 and NO2 at a concentration of not less than 50 mg / L, or not less than 100 mg / L, or not less than 200 mg / L. - , where each possibility represents a separate implementation.

[0247] According to aspects of the present invention, there is also provided a liquid composition comprising a biocompatible polymer, wherein the liquid composition undergoes a phase transition to a gel form upon stimulation. The liquid composition further comprises a reactive substance, at least one of which is selected from the group consisting of: OH * HO2 * 、O2 - 、O3、NO * 、NO3 - , H2O2, NO2 - and ONOO * , where reactive species have been produced by atmospheric plasma in arc mode.

[0248] According to another aspect of the present invention, there is provided a liquid composition comprising a synthetic biocompatible polymer, wherein the liquid composition undergoes a phase transition to a gel form upon stimulation. The liquid composition further comprises a reactive species, at least one of which is selected from the group consisting of: OH * HO2 * 、O2 - 、O3、NO * 、NO3 - , H2O2, NO2 - and ONOO * .

[0249] According to some embodiments, the stimulus of any of the above-mentioned liquid compositions comprises a temperature change. According to some embodiments, the stimulus comprises a pH change. According to some embodiments, the stimulus comprises light, and the phase change involves light-induced cross-linking.

[0250] According to some embodiments, the synthetic biocompatible polymer comprises polyethylene glycol (PEG), polypropylene glycol (PPG), poly(meth)acrylic acid, poly(meth)acrylate, or a combination thereof. According to some embodiments, the synthetic biocompatible polymer comprises a poloxamer. According to some embodiments, the poloxamer is selected from the group consisting of: poloxamer 101, poloxamer 105, poloxamer 108, poloxamer 122, poloxamer 123, poloxamer 124, poloxamer 181, poloxamer 182, poloxamer 183, poloxamer 184, poloxamer 185, poloxamer 188, poloxamer 212, poloxamer 215, poloxamer 217 , Poloxamer 231, Poloxamer 234, Poloxamer 235, Poloxamer 237, Poloxamer 238, Poloxamer 282, Poloxamer 284, Poloxamer 288, Poloxamer 331, Poloxamer 333, Poloxamer 334, Poloxamer 335, Poloxamer 338, Poloxamer 401, Poloxamer 402, Poloxamer 403, and Poloxamer 407, or mixtures thereof.

[0251] According to some embodiments, the liquid composition further comprises a fluid medium containing a buffer or a pH adjuster. According to some embodiments, the buffer or pH adjuster is selected from the group consisting of: 2-amino-2-hydroxymethyl-1,3-propanediol (Tris), 2-[bis(2-hydroxyethyl)imino]-2-(hydroxymethyl)-1,3-propanediol (bis-Tris), 4-morpholineethanesulfonic acid (MES) buffer, ammonium chloride, N,N-dihydroxyethylglycine, N-[tris(hydroxymethyl)methyl]glycine, sodium dihydrogen phosphate, sodium hydrogen phosphate, sodium carbonate, sodium bicarbonate, sodium acetate, sodium phosphate, glutamic acid, citrate buffer, histidine buffer In some embodiments, the liquid composition has a pH in the range of about 5.0 to about 7.5.

[0252] According to some embodiments, the liquid composition can be used to treat premalignant lesions. According to some embodiments, the liquid composition can be used to treat tumors. According to some embodiments, the liquid composition can be used to prevent or delay tumor recurrence after tumor resection. According to some embodiments, the liquid composition can be used to treat an infection selected from the group consisting of viral infections, bacterial infections, yeast infections, mold infections, and fungal infections.

[0253] According to another aspect of the present invention, there is provided a foam comprising reactive substances H2O2 and NO2 at a concentration of not less than 50 mg / Kg, or not less than 100 mg / Kg, or not less than 200 mg / Kg. - , wherein each possibility represents a separate embodiment. According to some embodiments, the foam further comprises a reactive substance OH * HO2 * 、O2 - 、O3、NO * 、NO3 - and ONOO * At least one of .

[0254] According to some embodiments, the foam is made from a liquid composition that is plasma activated and converted to foam by vigorous agitation. According to some embodiments, the liquid composition comprises a biocompatible polymer and is capable of undergoing a phase transition to a gel form upon stimulation. According to some embodiments, the stimulation comprises at least one selected from the group consisting of a temperature change, a pH change, and light-induced crosslinking.

[0255] According to a further aspect of the present invention, there is provided a method of treating an infection selected from viral infection, bacterial infection, yeast infection, mold infection and fungal infection in a subject in need thereof, the method comprising the step of topically administering to the subject a therapeutically effective amount of any of the liquid compositions described above or any foam produced therefrom.

[0256] According to some embodiments, the method further comprises, prior to the step of topically applying, plasma activating the liquid composition by forcing a flow of air onto the surface of the liquid composition while applying a plasma-generating EM field between an electrode external to the liquid composition and the surface of the liquid composition, thereby generating plasma in an arc mode along the air flow at ambient pressure.

[0257] According to some embodiments, the method further comprises agitating the liquid composition during said plasma activation. According to some embodiments, the method further comprises maintaining the liquid composition at a temperature below room temperature during said plasma activation. According to some embodiments, the temperature is below 10°C.

[0258] According to a further aspect of the present invention, there is provided a method of treating a premalignant lesion in a subject in need thereof, the method comprising the step of contacting the lesion with a therapeutically effective amount of any of the liquid compositions described above or any foam produced therefrom.

[0259] According to a further aspect of the present invention, there is provided a method of treating a tumor in a subject in need thereof, the method comprising the step of contacting the tumor with a therapeutically effective amount of any of the liquid compositions described above or any foam produced therefrom.

[0260] According to another aspect of the present invention, there is also provided a method for preventing or delaying tumor recurrence after tumor resection in a subject in need thereof, the method comprising the step of contacting the remaining tissue surrounding the tumor with a therapeutically effective amount of any liquid composition described above or any foam produced therefrom.

[0261] Although the present invention has been described in conjunction with specific embodiments thereof, it is obvious that many changes, modifications and variations will be apparent to those skilled in the art. It is therefore intended to embrace all such changes, modifications and variations that fall within the spirit and broad scope of the appended claims.

Claims

1. A system for providing a plasma activated medium (PAM) for medical use, the system comprising: a container configured to contain a liquid medium; an electrode electrically connected to a high voltage power source, the electrode being configured to apply an electromagnetic field to generate plasma in the container; an actuator configured to cause agitation of the liquid medium within the container, and a refrigerator having a cold portion, the refrigerator being configured to be thermally coupled to the liquid medium within the container, The system is configured to generate a plasma at atmospheric pressure within the container while agitating and maintaining the liquid medium at a temperature below room temperature.

2. The system according to claim 1, wherein the liquid medium is a liquid composition comprising a biocompatible polymer, wherein the liquid medium freezes at a temperature below T1 and accepts a gel form at a temperature above T2, wherein T1 <T2。 3. The system of claim 2, wherein T1 is between about -5°C and about +20°C, and T2 is between about +5°C and about +45°C.

4. The system of claim 3, wherein T1 is below about +15°C and T2 is above about +25°C.

5. The system of claim 2, wherein the biocompatible polymer is synthetic.

6. The system of claim 1, wherein the refrigerator comprises a vapor compression refrigerator.

7. The system of claim 6, wherein the refrigerator comprises a vortex tube.

8. The system of claim 1, wherein the refrigerator comprises a thermoelectric cooler.

9. The system of claim 1, wherein the refrigerator comprises an ice pack containing a coolant.

10. The system of claim 9, wherein the refrigerator is removable from the housing.

11. The system of claim 1, wherein the actuator and the HV power source are housed in a housing, and the container is removable from the housing.

12. The system of claim 1, wherein the walls of the container are dielectric.

13. The system of claim 1, wherein the walls of the container are metallic.

14. The system of claim 1, wherein the electrode is within the container.

15. The system of claim 1, wherein the electrode is shaped to have a tip, and the plasma is generated in an arc mode between the tip and the liquid medium.

16. The system of claim 1, further comprising a second electrode electrically connected to a second high voltage power source and configured to thereby apply a plasma-generating electromagnetic field within the vessel.

17. The system of claim 1, wherein the actuator is a stirrer and the agitation is achieved by stirring the liquid medium.

18. The system of claim 17, wherein the stirrer is a magnetic stirrer and the container comprises a magnetic plate.

19. The system of claim 1, wherein the actuator is a vibration actuator configured to vibrate the container, thereby shaking the liquid medium.

20. The system of claim 1, wherein the actuator is a compressor configured to force a gas flow onto the liquid medium, thereby agitating the liquid medium.

21. The system of claim 20, wherein the gas is released within the liquid medium in the container, thereby generating bubbles in the liquid medium.

22. The system of claim 1, further comprising a compressor fluidly connected to the container via an inlet passage and a gas inlet port of the container, the compressor configured to force gas into the container.

23. The system of claim 22, wherein the gas is air drawn from the environment.

24. The system of claim 22, wherein the compressor draws the gas from the container via an outlet passage fluidly connected to a gas outlet port of the container, thereby circulating the gas through the container.

25. The system of claim 22, wherein a plasma-generating EM field is applied to the gas flow.

26. The system of claim 25, wherein the electrode is shaped as an open-ended tube in fluid connection with the compressor, whereby the plasma-generating EM field is generated along the gas flow between a distal end of the tube and the liquid medium.

27. The system of claim 22, wherein the inlet passage comprises a filter.

28. The system of claim 24, wherein the outlet passage comprises a filter.

29. A transportable closed container comprising: a liquid medium, stored in said container, intended for medical use, and an electrode electrically connected to the HV connector, the electrode being configured to apply a plasma-generating EM field within the container upon receiving a high voltage via the HV connector, The container is bio-sealed to prevent bacteria or viruses from penetrating into the container.

30. The container of claim 29, wherein the walls of the container are dielectric.

31. The container of claim 29, wherein the walls of the container are metallic.

32. The container of claim 29 having a volume between 5 cc and 50 cc.

33. The container of claim 29, wherein the liquid composition is sterile.

34. The container of claim 29, further comprising an inlet port configured to enable a gas to flow into the container.

35. The vessel of claim 34, wherein the electrode comprises a metal tube in flow communication with the inlet port and having a distal portion within the vessel.

36. The container of claim 35, wherein the metal tube has a pointed tip at its distal end.

37. The container of claim 29, further comprising a magnetic plate immersed in the liquid composition and configured to cause agitation of the liquid when subjected to a corresponding magnetic field.

38. The container of claim 29, further comprising an outlet port that allows gas to flow out of the container.

39. The container of claim 29, further comprising a removable cover, the inlet port and HV connector being assembled to the cover.

40. The container of claim 29, wherein the liquid medium comprises a biocompatible polymer that forms a liquid composition capable of undergoing a phase transition to a gel form upon stimulation.

41. A portable, passive refrigerator configured to refrigerate a container, the passive refrigerator comprising an ice pack having at least one chamber for containing a coolant having a freezing temperature between -10°C and +5°C, the at least one chamber being shaped around a groove in the ice pack, the groove being configured and sized to receive the container therein.

42. The passive cooler of claim 41 further comprising the container of the ice pack attached to the trough, the container being metallic and in direct contact with the coolant.

43. The passive cooler of claim 42, further comprising an openable cover for the container.

44. The passive cooler of claim 42, further comprising an HV connector and an electrode electrically connected to the HV connector and electrically insulated from the metal container, the electrode being configured to apply a plasma-generating EM field in the container when receiving a high voltage from a HV power source.

45. The passive cooler of claim 41, wherein the grooves are through holes in the ice pack.

46. ​​The passive cooler of claim 41, wherein the groove is formed as a depression in the ice pack.

47. The passive cooler of claim 41 further comprising a non-rigid member surrounding the groove, the non-rigid member being configured to adjustably contact the container when the container is placed in the groove to establish thermal contact between the ice pack and the container.

48. The passive cooler of claim 47, wherein the non-rigid member comprises a flexible metal portion.

49. The passive refrigerator of claim 47, wherein the non-rigid member comprises at least one second chamber, at least one wall of the at least one second chamber being non-rigid, the at least one second chamber containing a material that is liquid at a freezing temperature of the coolant.

50. The passive cooler of claim 49, wherein at least one of the non-rigid walls is soft.

51. A liquid composition comprising a biocompatible polymer, wherein the liquid composition undergoes a phase transition to a gel form upon stimulation, and the liquid composition further comprises reactive substances H2O2 and NO2 at a combined concentration of not less than 100 mg / L - .

52. The liquid composition of claim 51, wherein the reactive species has been generated by atmospheric plasma in arc mode.

53. The liquid composition of claim 51, wherein the biocompatible polymer is synthetic.

54. The liquid composition according to claim 51, wherein the liquid composition further comprises OH * HO2 * 、O2 - 、O3、NO * 、NO3 - and ONOO * At least one of .

55. The liquid composition of any one of claims 51 to 54, wherein the stimulus comprises a temperature change.

56. The liquid composition of any one of claims 51 to 54, wherein the stimulus comprises a pH change.

57. The liquid composition of any one of claims 51 to 54, wherein the stimulus comprises light and the phase change involves light-induced cross-linking.

58. The liquid composition of claim 53, wherein the synthetic biocompatible polymer comprises polyethylene glycol (PEG), polypropylene glycol (PPG), poly(meth)acrylic acid, poly(meth)acrylate, or a combination thereof.

59. The liquid composition of claim 58, wherein the synthetic biocompatible polymer comprises a poloxamer.

60. The liquid composition of claim 59, wherein the poloxamer is selected from the group consisting of poloxamer 101, poloxamer 105, poloxamer 108, poloxamer 122, poloxamer 123, poloxamer 124, poloxamer 181, poloxamer 182, poloxamer 183, poloxamer 184, poloxamer 185, poloxamer 188, poloxamer 212, poloxamer 215, Poloxamer 217, poloxamer 231, poloxamer 234, poloxamer 235, poloxamer 237, poloxamer 238, poloxamer 282, poloxamer 284, poloxamer 288, poloxamer 331, poloxamer 333, poloxamer 334, poloxamer 335, poloxamer 338, poloxamer 401, poloxamer 402, poloxamer 403, and poloxamer 407 or mixtures thereof.

61. The liquid composition of any one of claims 51 to 60, wherein the composition further comprises a fluid medium comprising a buffer or a pH adjuster.

62. The liquid composition of claim 61, wherein the buffer or pH adjuster is selected from the group consisting of 2-amino-2-hydroxymethyl-1,3-propanediol (Tris), 2-[bis(2-hydroxyethyl)imino]-2-(hydroxymethyl)-1,3-propanediol (bis-Tris), 4-morpholineethanesulfonic acid (MES) buffer, ammonium chloride, N,N-bicine, N-[tris(hydroxymethyl)methyl]glycine, sodium dihydrogen phosphate, sodium hydrogen phosphate, sodium carbonate, sodium bicarbonate, sodium acetate, sodium phosphate, glutamic acid, citrate buffer, , histidine buffer, Dulbecco's phosphate buffered saline, 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES), methoxypsoralen (MOPS), N-cyclohexyl-3-aminopropanesulfonic acid (CAPS), N-cyclohexyl-2-hydroxy-3-aminopropanesulfonic acid (CAPSO), N-cyclohexyl-2-aminoethanesulfonic acid (CHES), 3-[4-(2-hydroxyethyl)-1-piperazinyl]propanesulfonic acid (HEPPS), phosphate buffered saline, tris buffered saline, Hank's solution and Ringer's solution, and mixtures or combinations thereof.

63. The liquid composition of any one of claims 51 to 62, having a pH in the range of about 5.0 to about 7.

5.

64. A liquid composition according to any one of claims 51 to 63 for use in the treatment of premalignant lesions.

65. A liquid composition according to any one of claims 51 to 63 for use in treating a tumour.

66. The liquid composition according to any one of claims 51 to 63, for use in preventing or delaying tumor recurrence after tumor resection.

67. A liquid composition according to any one of claims 51 to 63 for use in treating an infection selected from a viral infection, a bacterial infection, a yeast infection, a mold infection and a fungal infection.

68. A foam comprising reactive substances H2O2 and NO2 in concentrations not less than 100 mg / kg - .

69. The foam according to claim 68, further comprising a reactive substance OH * HO2 * 、O2 - 、O3、NO * 、NO3 - and ONOO * At least one of .

70. The foam of claim 68, made from a liquid composition that has been plasma activated and converted to foam by intense agitation.

71. The foam of claim 68, wherein the liquid composition comprises a biocompatible polymer and is capable of undergoing a phase transition to a gel form upon stimulation.

72. The foam of claim 68, wherein the stimulus comprises at least one from the group consisting of temperature change, pH change, and light-induced cross-linking.

73. A method of treating an infection selected from the group consisting of a viral infection, a bacterial infection, a yeast infection, a mold infection and a fungal infection in a subject in need thereof, the method comprising the step of topically administering to the subject a therapeutically effective amount of a liquid composition or foam according to any one of claims 51 to 72.

74. The method of claim 73, further comprising, prior to the step of topically applying, plasma activating the liquid composition by forcing a flow of air onto a surface of the liquid composition while applying a plasma-generating EM field between an electrode external to the liquid composition and the surface of the liquid composition, thereby generating plasma in an arc mode along the air flow at ambient pressure.

75. The method of claim 74, further comprising agitating the liquid composition during the plasma activation.

76. The method of claim 74, further comprising maintaining the liquid composition at a temperature below room temperature during the plasma activation.

77. The method of claim 76, wherein the temperature is below 10°C.

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