A method of storing plasma active particles and products

Abstract

This invention relates to the field of plasma active particle storage technology, specifically to a method and product for storing plasma active particles. The method includes: plasma-activating a polymer solution to obtain a plasma-activated polymer solution, the plasma-activated polymer solution comprising plasma active particles; adding a gel precursor solution to the plasma-activated polymer solution according to a preset ratio to obtain a mixed solution, the gel precursor solution comprising at least one of a gelatin solution and a polyvinyl alcohol solution; and cross-linking the mixed solution to obtain a composite gel loaded with plasma active particles. This invention, by plasma-activating the polymer solution, allows plasma-generated active particles to enter the polymer system and combine with gel cross-linking to form a three-dimensional network structure, achieving the loading and stable storage of active particles, thereby effectively extending the lifetime of the active particles.

Description

Technical Field

[0001] This invention relates to the field of plasma active particle storage technology, and specifically to a method and product for storing plasma active particles. Background Technology

[0002] Low-temperature plasma has broad application prospects in fields such as biomedicine, wound repair, antibacterial disinfection, tumor treatment, and tissue engineering. During plasma interaction, various reactive particles are generated, including reactive oxygen species, reactive nitrogen species, free radicals, excited-state molecules, and ions. These reactive particles can participate in redox reactions, cell signaling regulation, and microbial inactivation processes, thereby endowing plasma with excellent biological activity and therapeutic effects.

[0003] Currently, plasma-generated active particles typically act directly on the target area in gaseous form. Polymers such as hyaluronic acid possess excellent biocompatibility, biodegradability, and tissue compatibility, showing broad application prospects in the field of biomedical materials. Plasma can be used to temporarily store and transfer active particles through activation treatment of polymer systems such as hyaluronic acid. However, these polymers, in their uncrosslinked state, are usually in solution form, limiting their ability to store plasma-generated active particles. They are particularly unable to effectively maintain the stability of short-lived and some long-lived active particles, leading to rapid activity decay. Summary of the Invention

[0004] (a) Purpose of the invention The purpose of this invention is to provide a method and product for storing plasma-generated active particles. By subjecting a polymer solution to plasma activation treatment, plasma-generated active particles are introduced into the polymer system and further combined with gel crosslinking to form a three-dimensional network structure. This achieves the loading and stable storage of active particles, thereby effectively extending their lifespan. Furthermore, due to the thermosensitive properties of gelatin, the active particles can be released controllably through temperature regulation, further enhancing the flexibility and adaptability of this technology.

[0005] (II) Technical Solution To address the above problems, the present invention provides a method for storing plasma active particles, comprising: A polymer solution is subjected to plasma activation treatment to obtain a plasma-activated polymer solution, wherein the plasma-activated polymer solution includes plasma-active particles; The gel precursor solution is added to the plasma-activated polymer solution according to a preset ratio and mixed to obtain a mixed solution. The gel precursor solution includes at least one of gelatin solution and polyvinyl alcohol solution, and the gel precursor solution is acidic. The mixed solution was subjected to cross-linking treatment to obtain a composite gel loaded with plasma-active particles.

[0006] In another aspect of the present invention, preferably, the polymer in the polymer solution includes at least one of hyaluronic acid, sodium hyaluronate, alginate, chitosan, cellulose, cellulose derivatives, pectin, starch, and dextran.

[0007] In another aspect of the present invention, preferably, the apparatus used for the plasma activation treatment includes one or more of the following: a plasma jet apparatus, a dielectric barrier discharge apparatus, a sliding arc discharge apparatus, and a spark discharge apparatus; The plasma activation treatment methods include irradiation, air blowing, bubbling, and a combination of irradiation and air blowing. The plasma irradiation activation treatment is carried out in an ambient gas atmosphere, and the working gas introduced for the gas blowing or bubbling activation treatment includes at least one of air, nitrogen, oxygen, argon and helium. The working gas flow rate for the plasma blowing or bubbling activation treatment is 0.1 SLM to 5 SLM. The plasma activation treatment time is 5 min to 90 min.

[0008] In another aspect of the present invention, preferably, the concentration of the gelatin solution is 5% to 40% (w / v), the temperature of the gelatin solution is 37°C to 50°C, and the concentration of the polyvinyl alcohol is 5% to 25% (w / v).

[0009] In another aspect of the present invention, preferably, the pH value of the gel precursor solution is 3 to 4; when the gel precursor solution includes the gelatin solution and the polyvinyl alcohol solution, the volume ratio of the gelatin solution and the polyvinyl alcohol solution is 1:0.2 to 5.

[0010] In another aspect of the present invention, preferably, the concentration of the polymer in the plasma-activated polymer solution is 0.5% to 2% (w / v). The preset ratio is the volume ratio of the plasma-activated polymer solution to the gel precursor solution, and the preset ratio is 1 to 5:1.

[0011] In another aspect of the present invention, preferably, the volume ratio of the gelatin solution to the polyvinyl alcohol solution is 1:0.5 to 2; The preset ratio is 1 to 2:1.

[0012] In another aspect of the invention, preferably, when the gel precursor solution comprises the gelatin solution and the polyvinyl alcohol solution, the crosslinking treatment comprises a cooling treatment and / or a freeze-thaw treatment; When the gel precursor solution includes a gelatin solution, the crosslinking treatment includes a cooling treatment; When the gel precursor solution includes a polyvinyl alcohol solution, the crosslinking treatment includes a freeze-thaw process; The cooling process includes cooling the mixed solution to 25°C or below and maintaining it for 5 min to 90 min; The freeze-thaw treatment includes freezing the mixed solution at -20℃ to -80℃ for 8h to 16h and thawing it at 20℃ to 40℃ for 4h to 8h. The freeze-thaw treatment is repeated 1 to 5 times.

[0013] In another aspect of the present invention, preferably, the method further includes: The composite gel is freeze-dried to obtain a freeze-dried composite gel product, which includes: pre-freezing the composite gel at -20℃ to -80℃, and freeze-drying the pre-frozen composite gel under vacuum conditions to obtain a freeze-dried composite gel product. The composite gel and / or the freeze-dried composite gel product shall be stored under low temperature, light-proof and sealed conditions.

[0014] In another aspect of the present invention, preferably, a product loaded with plasma-active particles, the product comprising a composite gel and / or a lyophilized composite gel product, wherein the composite gel is prepared using the method described above, and the lyophilized composite gel product is prepared using the method described above.

[0015] (III) Beneficial Effects The above-described technical solution of the present invention has the following beneficial technical effects: This invention achieves effective loading and storage of plasma-generated active particles by combining a plasma-activated polymer solution with a gel precursor solution and cross-linking them to form a composite gel. Gelatin rapidly forms a physical gel network upon cooling, while PVA forms a stable physical cross-linked network after freeze-thaw cycles. The combination of these two further forms a composite network structure, thereby spatially confining the active particles and reducing their diffusion rate, recombination, quenching, and decomposition probabilities. Simultaneously, the intermolecular interactions and network interpenetration between PVA and gelatin help reduce the consumption of active particles at some reaction sites. Furthermore, the resulting composite gel exhibits thermosensitivity, enhancing its fluidity and accelerating the release of active particles under specific temperature conditions.

[0016] Furthermore, the composite gel is freeze-dried to obtain freeze-dried composite gel products that are easy to store and transport. Freeze-dried composite gel products can expand application forms and scenarios. For example, freeze-dried composite gel products can be applied directly to the target area, or they can be rehydrated with a rehydration solution before application. Composite gels and freeze-dried composite gel products can be used for sterilization, anti-infection, wound covering, wound management, and related dressing scenarios. Attached Figure Description

[0017] Figure 1 This is an overall flowchart of one embodiment of the present invention. Detailed Implementation

[0018] To make the objectives, technical solutions, and advantages of this invention clearer, the invention will be further described in detail below with reference to specific embodiments and the accompanying drawings. It should be understood that these descriptions are merely exemplary and not intended to limit the scope of the invention. Furthermore, descriptions of well-known structures and techniques are omitted in the following description to avoid unnecessarily obscuring the concept of the invention.

[0019] Obviously, the described embodiments are only some, not all, of the embodiments of the present invention. All other embodiments obtained by those skilled in the art based on the embodiments of the present invention without inventive effort are within the scope of protection of the present invention.

[0020] Furthermore, the technical features involved in the different embodiments of the present invention described below can be combined with each other as long as they do not conflict with each other.

[0021] Example A method for storing plasma-active particles. Figure 1 An overall flowchart of one embodiment of the present invention is shown, as follows: Figure 1 As shown, it includes: A polymer solution is subjected to plasma activation treatment to obtain a plasma-activated polymer solution, wherein the plasma-activated polymer solution includes plasma active particles; the active particles include active oxygen particles and / or active nitrogen particles, specifically including one or more of hydroxyl radicals, superoxide anions, singlet oxygen, hydrogen peroxide, ozone, nitric oxide, nitrate ions, and nitrite ions. Through plasma activation treatment, a certain concentration of active particles can be loaded into the polymer solution, forming a relatively stable activation system.

[0022] Furthermore, in this embodiment, the polymer in the polymer solution includes at least one of hyaluronic acid, sodium hyaluronate, alginate, chitosan, cellulose, cellulose derivatives, pectin, starch, and dextran. All of these polymers possess good hydrophilicity and biocompatibility; some also exhibit thickening, water-retaining, or gelling properties, enabling them to form a stable polymer molecular chain entanglement environment in the liquid system, thereby providing a carrier basis for the storage of plasma active particles. Different polymers can also be combined according to actual needs to adjust the viscosity, water retention, mechanical strength, and sustained-release performance of the system.

[0023] Furthermore, in this embodiment, the polymer solution can be placed within the effective area of ​​the plasma generator. Low-temperature plasma is generated through dielectric barrier discharge or jet plasma discharge, allowing the active particles generated by the plasma to enter the polymer solution. Alternatively, the activation gas generated by the plasma can be transported to the surface or near-surface area of ​​the polymer solution for gas blowing treatment, or the activation gas generated by the plasma can be directly introduced into the polymer solution for bubbling treatment. The device used for plasma activation treatment includes one or more of the following: plasma jet device, dielectric barrier discharge device, sliding arc discharge device, and spark discharge device; different discharge devices can be selected according to actual application requirements. Specifically, the plasma jet device can stably generate low-temperature plasma under normal pressure and achieve directional treatment of local areas; the dielectric barrier discharge device has the characteristics of uniform discharge, low temperature rise, and suitability for large-area treatment; the sliding arc discharge device can generate a high concentration of active particles; and the spark discharge device has the characteristics of high instantaneous energy and high active particle generation efficiency. By selecting different discharge devices, the type, concentration, and generation efficiency of active particles can be adjusted to meet the needs of different polymer systems and application scenarios.

[0024] The plasma activation treatment methods include irradiation, gas blowing, bubbling, and a combination of irradiation and gas blowing. Irradiation refers to placing the plasma discharge area above or near the surface of the polymer solution, treating the surface of the polymer solution through radiation, electric field, and active particles generated by the plasma. Gas blowing refers to delivering the activation gas generated by the plasma to the surface or near the liquid surface of the polymer solution, allowing active particles to enter the polymer solution through the gas-liquid interface by purging. Bubbling refers to directly introducing the activation gas generated by the plasma into the interior of the polymer solution, causing it to disperse and rise in the liquid as bubbles, thereby achieving the transfer of active particles into the polymer solution. The combination of irradiation and gas blowing refers to simultaneously irradiating the polymer solution with a plasma plume and blowing the polymer solution with an incoming working gas carrying active particles.

[0025] The plasma irradiation activation treatment is carried out in an ambient gas atmosphere, including ambient air. The working gas introduced for the air blowing or bubbling activation treatment includes at least one of air, nitrogen, oxygen, argon, and helium. Air can economically and efficiently generate active oxygen and active nitrogen particles; oxygen is beneficial for generating active oxygen particles; nitrogen is beneficial for generating active nitrogen particles; argon and helium, as inert gases, can improve discharge stability and reduce the thermal effect during the treatment. Different working gases can also be mixed to adjust the composition and concentration of active particles.

[0026] The working gas flow rate for the plasma blowing or bubbling activation treatment is 0.1 SLM to 5 SLM; specifically, it can be 0.1 SLM, 0.5 SLM, 1 SLM, 1.5 SLM, 2 SLM, 2.5 SLM, 3 SLM, 3.5 SLM, 4 SLM, 4.5 SLM, or 5 SLM, etc., to adjust the discharge stability, active particle transport efficiency, and gas-liquid mass transfer efficiency.

[0027] The plasma activation treatment time is 5 min to 90 min, specifically 5 min, 10 min, 15 min, 20 min, 40 min, 60 min, 70 min, 80 min and 90 min, etc., which can be set according to different polymer systems and the required concentration of target active particles.

[0028] The gel precursor solution is added to the plasma-activated polymer solution according to a preset ratio to obtain a mixed solution. The gel precursor solution includes at least one of gelatin solution and polyvinyl alcohol solution. In this embodiment, the gel precursor solution is acidic, with a pH value of 3-4, specifically 3, 3.2, 3.5, 3.7, 3.8, or 4. The pH of the gel precursor solution can be adjusted using a diluted acid solution, including at least one of hydrochloric acid and nitric acid, which can be diluted with water, deionized water, physiological saline, or PBS buffer. Acidic conditions are beneficial for maintaining the bioactivity of the plasma-activated system and can slow down the decay of some active particles. At the same time, a suitable acid range also helps to balance the stability of active particles and the quality of gel formation. If the pH value is too low, it may lead to excessive hydrolysis of gelatin molecular chains or a decrease in the stability of the polyvinyl alcohol system, affecting the quality of gel formation. If the pH value is too high, it may accelerate the decay of some active particles, which is not conducive to maintaining the bioactivity of the plasma-activated system.

[0029] The plasma activation system is brought into full contact with the gel precursor solution, thereby achieving effective encapsulation and confinement of active particles during the gel formation process. The concentration of the gelatin solution is 5%–40% (w / v), specifically 5%, 10%, 15%, 20%, 25%, 30%, 35%, or 40%, etc. The temperature of the gelatin solution is 37℃–50℃, specifically 37℃, 39℃, 41℃, 43℃, 45℃, 47℃, 49℃, or 50℃, etc., to provide stable temperature-sensitive gelling ability. The concentration of the polyvinyl alcohol is 5%–25% (w / v), specifically 5%, 7%, 10%, 12%, 14%, 16%, 18%, 20%, 22%, or 25%, etc., to form a stable cross-linked network after freeze-thaw cycles. When the gel precursor solution includes the gelatin solution and the polyvinyl alcohol solution, the volume ratio of the gelatin solution and the polyvinyl alcohol solution is 1:0.2 to 5, specifically 1:0.2, 1:0.5, 1:1, 1:2, 1:3, 1:4 or 1:5, etc., to take into account both the thermosensitive properties and freeze-thaw properties of the prepared composite gel.

[0030] The concentration of polymer in the plasma-activated polymer solution is 0.5%–2% (w / v), specifically 0.5%, 0.8%, 1.0%, 1.5%, or 2%, etc., which has good solution stability and biocompatibility, and is conducive to the loading of plasma active particles into the liquid phase and the subsequent formation of composite gel. The preset ratio is the volume ratio of plasma-activated polymer solution to gel precursor solution, and the preset ratio is 1–5:1, specifically 1:1, 1.5:1, 2:1, 2.5:1, 3:1, 3.5:1, 4:1, 4.5:1, or 5:1, etc., to ensure that the polymer loaded with active particles accounts for a sufficient proportion in the prepared composite gel, and to achieve a better balance between temperature-sensitive gelation performance and active particle storage performance.

[0031] The mixed solution is subjected to crosslinking treatment to obtain a composite gel loaded with plasma active particles. When the gel precursor solution includes the gelatin solution and the polyvinyl alcohol solution, the crosslinking treatment includes cooling treatment and / or freeze-thaw treatment. When the gel precursor solution includes a gelatin solution, the crosslinking treatment includes a cooling treatment; When the gel precursor solution includes a polyvinyl alcohol solution, the crosslinking treatment includes a freeze-thaw process; The cooling process includes cooling the mixed solution to 25°C or below, specifically 25°C, 20°C, 15°C, 10°C, 4°C, or 0°C, and maintaining this temperature for 5 min to 90 min, specifically 5 min, 10 min, 20 min, 30 min, 40 min, 50 min, 60 min, 70 min, 80 min, or 90 min. This facilitates the rapid formation and uniform stability of the gelatin network, thereby improving the uniformity and structural integrity of the resulting composite gel and reducing the loss of active particles.

[0032] The freeze-thaw treatment includes freezing the mixed solution at -20℃ to -80℃ (specifically -20℃, -30℃, -40℃, -50℃, -60℃, -70℃, or -80℃) for 8h to 16h (specifically 8h, 10h, 11h, 12h, 13h, 14h, 15h, or 16h); and thawing the solution at 20℃ to 40℃ (specifically 20℃, 25℃, 30℃, 35℃, or 40℃) for 4h to 8h (specifically 4h, 5h, 6h, 7h, or 8h). The freeze-thaw treatment cycle is 1 to 5 times (specifically 1, 2, 3, 4, or 5 times) to enhance the network strength of the composite gel and improve the storage capacity of active particles.

[0033] Furthermore, in this embodiment, the method further includes: The composite gel is freeze-dried to obtain a freeze-dried composite gel product; freeze-drying improves the storage and transportation convenience of the composite gel. Storing the freeze-dried composite gel product under low temperature, light-proof, and sealed conditions extends the lifespan of the active particles.

[0034] The process of freeze-drying the composite gel to obtain a freeze-dried composite gel product includes: placing the composite gel under conditions of -20℃ to -80℃, specifically -20℃, -30℃, -40℃, -50℃, -60℃, -70℃ or -80℃ for pre-freezing; and freeze-drying the pre-frozen composite gel under vacuum conditions to obtain a freeze-dried composite gel product. In this embodiment, the composite gel and / or freeze-dried composite gel product can be stored under low temperature, light-proof and sealed conditions to reduce the attenuation of active particles during storage.

[0035] In this embodiment, the freeze-dried composite gel product can be applied directly to the target area, or it can be rehydrated with a rehydration solution before application; the rehydration solution includes at least one of water, deionized water, physiological saline and PBS buffer.

[0036] This embodiment introduces gelatin and polyvinyl alcohol into a plasma-activated polymer solution, forming a composite gel through mixing and physical crosslinking. A gel network for storing plasma-activated particles can be constructed without a complex chemical crosslinking process. The physical network formed by gelatin and / or polyvinyl alcohol can confine and slow-release the activated particles, thereby reducing their diffusion, recombination, and decay rates, and improving the system's overall storage capacity for activated particles with different lifespans. The gelatin network, the PVA freeze-thaw network, and their combined network exhibit different retention effects on activated particles with different lifespans; combining them further enhances the composite gel's overall storage capacity for multiple types of activated particles. Furthermore, the resulting composite gel system containing gelatin also exhibits temperature sensitivity, with significantly enhanced fluidity under heating conditions, thus accelerating the release of activated particles. Moreover, the composite gel can be freeze-dried to form freeze-dried composite gel products, thereby improving the material's transport convenience, storage stability, and application flexibility.

[0037] This embodiment also provides a product loaded with plasma-active particles, characterized in that the product comprises a composite gel and / or a lyophilized composite gel product, wherein the composite gel is prepared using the method described above, the lyophilized composite gel product is prepared using the method described above, and the composite gel or lyophilized composite gel product contains a plasma-activated polymer component and a gelatin and / or polyvinyl alcohol component.

[0038] Example 1 A 1% (w / v) hyaluronic acid solution was prepared and subjected to plasma irradiation activation treatment using a plasma dielectric barrier discharge device. The plasma irradiation activation treatment was carried out in an ambient air atmosphere for 60 minutes. The active particles generated by the plasma were then loaded into the hyaluronic acid solution to obtain a plasma-activated hyaluronic acid solution.

[0039] Prepare a 20% (w / v) gelatin solution and a 15% (w / v) polyvinyl alcohol solution, with the gelatin solution temperature controlled at 40℃. Mix the liquid gelatin solution and polyvinyl alcohol solution evenly at a volume ratio of 1:1. Dilute the hydrochloric acid with physiological saline to adjust the pH to 3.5 to obtain the gel precursor solution.

[0040] The plasma-activated hyaluronic acid solution and the gel precursor solution were mixed evenly at a volume ratio of 3:2 to obtain a mixed solution. Finally, the mixed solution was cooled at 4°C for 30 min, then frozen at -20°C for 12 h, and then thawed at 25°C for 4 h. The above freeze-thaw cycle was repeated 3 times to obtain the composite gel.

[0041] Example 2 A 1% (w / v) hyaluronic acid solution was prepared and subjected to plasma irradiation activation treatment using a plasma dielectric barrier discharge device. The plasma irradiation activation treatment was carried out in an ambient air atmosphere for 60 minutes. The active particles generated by the plasma were then loaded into the hyaluronic acid solution to obtain a plasma-activated hyaluronic acid solution.

[0042] Prepare a 10% (w / v) gelatin solution at a temperature of 40°C. Dilute the hydrochloric acid with physiological saline to achieve a pH of 3.5 to obtain the gel precursor solution.

[0043] The plasma-activated hyaluronic acid solution and the gel precursor solution were mixed thoroughly at a volume ratio of 3:2 to obtain a mixed solution. Finally, the mixed solution was cooled at 4°C for 30 min to obtain a composite gel.

[0044] Example 3 A 1% (w / v) hyaluronic acid solution was prepared and subjected to plasma irradiation activation treatment using a plasma dielectric barrier discharge device. The plasma irradiation activation treatment was carried out in an ambient air atmosphere for 60 minutes. The active particles generated by the plasma were then loaded into the hyaluronic acid solution to obtain a plasma-activated hyaluronic acid solution.

[0045] Prepare a 7.5% (w / v) polyvinyl alcohol solution, dilute hydrochloric acid with physiological saline to make the pH 3.5, and obtain the gel precursor solution.

[0046] The plasma-activated hyaluronic acid solution and the gel precursor solution were mixed evenly at a volume ratio of 3:2 to obtain a mixed solution. Finally, the mixed solution was frozen at -20°C for 12 hours and then thawed at 25°C for 4 hours. This freeze-thaw cycle was repeated three times to obtain the composite gel.

[0047] Example 4 A 1% (w / v) hyaluronic acid solution was prepared and subjected to plasma irradiation activation treatment using a plasma dielectric barrier discharge device. The plasma irradiation activation treatment was carried out in an ambient air atmosphere for 60 minutes. The active particles generated by the plasma were then loaded into the hyaluronic acid solution to obtain a plasma-activated hyaluronic acid solution.

[0048] Prepare a 20% (w / v) gelatin solution and a 15% (w / v) polyvinyl alcohol solution, with the gelatin solution temperature controlled at 40℃. Mix the liquid gelatin solution and polyvinyl alcohol solution evenly at a volume ratio of 1:1. Dilute the hydrochloric acid with physiological saline to adjust the pH to 3.5 to obtain the gel precursor solution.

[0049] The plasma-activated hyaluronic acid solution and the gel precursor solution were mixed evenly at a volume ratio of 3:2 to obtain a mixed solution. Finally, the mixed solution was cooled at 4°C for 30 min, then frozen at -20°C for 12 h, and then thawed at 25°C for 4 h. The above freeze-thaw cycle was repeated 3 times to obtain the composite gel.

[0050] The composite gel was pre-frozen at -80°C. The pre-frozen composite gel was freeze-dried under vacuum conditions to obtain the freeze-dried composite gel product.

[0051] The freeze-dried composite gel product can be stored under low temperature, light-proof and sealed conditions.

[0052] Example 5 A 0.5% (w / v) chitosan solution was prepared and subjected to plasma gas blowing treatment using a plasma sliding arc discharge device. The working gas for plasma activation treatment was air, the working gas flow rate was 5 SLM, and the plasma activation treatment time was 90 min. The active particles generated by the plasma were then loaded into the chitosan solution to obtain a plasma-activated chitosan solution.

[0053] Prepare a 5% (w / v) gelatin solution and a 20% (w / v) polyvinyl alcohol solution, with the gelatin solution temperature controlled at 37°C. Mix the liquid gelatin solution and polyvinyl alcohol solution evenly at a volume ratio of 1:0.2, and dilute the hydrochloric acid with deionized water to prepare a pH of 3 to obtain the gel precursor solution.

[0054] The plasma-activated chitosan solution and the gel precursor solution were mixed evenly at a volume ratio of 1:1 to obtain a mixed solution. Finally, the mixed solution was cooled at 25°C for 5 min, then frozen at -80°C for 8 h, and then thawed at 20°C for 8 h. This freeze-thaw cycle was repeated once to obtain the composite gel.

[0055] Example 6 A 2% (w / v) cellulose solution was prepared and subjected to plasma activation treatment using a plasma jet device. The plasma activation treatment was carried out by a combination of irradiation and gas blowing. The irradiation activation treatment was carried out in an ambient air atmosphere, and the working gas introduced for the gas blowing activation treatment was helium. The flow rate of the helium was 1 slm, and the plasma activation treatment time was 60 min. The active particles generated by the plasma were then loaded into the cellulose solution to obtain a plasma-activated cellulose solution.

[0056] Prepare a 40% (w / v) gelatin solution and a 5% (w / v) polyvinyl alcohol solution, with the gelatin solution temperature controlled at 50°C. Mix the liquid gelatin solution and polyvinyl alcohol solution evenly at a volume ratio of 1:5, and dilute the nitric acid with deionized water to prepare a pH of 4 to obtain the gel precursor solution.

[0057] The plasma-activated cellulose solution and the gel precursor solution were mixed evenly at a volume ratio of 5:1 to obtain a mixed solution. Finally, the mixed solution was cooled at 0°C for 90 min, then frozen at -30°C for 16 h, and then thawed at 40°C for 6 h. The above freeze-thaw cycle was repeated 5 times to obtain the composite gel.

[0058] Example 7 A 1.5% (w / v) pectin solution was prepared using a plasma spark discharge device. The plasma activation treatment was performed by bubbling, with air as the working gas and a flow rate of 0.1 SLM. The plasma activation treatment time was 45 min. The active particles generated by the plasma were then loaded into the pectin solution to obtain a plasma-activated pectin solution.

[0059] Prepare a 15% (w / v) gelatin solution and a 12% (w / v) polyvinyl alcohol solution, with the gelatin solution temperature controlled at 42℃. Mix the liquid gelatin solution and polyvinyl alcohol solution evenly at a volume ratio of 1:0.5, and dilute the nitric acid with deionized water to prepare a pH of 3.8 to obtain the gel precursor solution.

[0060] The plasma-activated pectin solution and the gel precursor solution were mixed evenly at a volume ratio of 2:1 to obtain a mixed solution. Finally, the mixed solution was cooled at 10°C for 80 min, then frozen at -40°C for 12 h, and then thawed at 30°C for 7 h. The above freeze-thaw cycle was repeated 4 times to obtain the composite gel.

[0061] Example 8 A 0.8% (w / v) starch solution was prepared using a plasma dielectric barrier discharge device. The plasma activation treatment was performed by bubbling, with a working gas mixture of nitrogen and oxygen at a flow rate of 3 SLM and a treatment time of 5 min. The active particles generated by the plasma were then loaded into the starch solution to obtain a plasma-activated starch solution.

[0062] Prepare a 17% (w / v) gelatin solution and a 25% (w / v) polyvinyl alcohol solution, with the gelatin solution temperature controlled at 45℃. Mix the liquid gelatin solution and polyvinyl alcohol solution evenly at a volume ratio of 1:3, and dilute the nitric acid with deionized water to prepare a pH of 3.2 to obtain the gel precursor solution.

[0063] The plasma-activated starch solution and the gel precursor solution were mixed evenly at a volume ratio of 3:1 to obtain a mixed solution. Finally, the mixed solution was cooled at 8°C for 60 min, then frozen at -35°C for 14 h, and then thawed at 35°C for 5 h. The above freeze-thaw cycle was repeated twice to obtain the composite gel.

[0064] Comparative Example 1 A 1% (w / v) hyaluronic acid solution was prepared and subjected to plasma irradiation activation treatment using a plasma dielectric barrier discharge device. The plasma irradiation activation treatment was carried out in an ambient air atmosphere for 60 minutes. The active particles generated by the plasma were then loaded into the hyaluronic acid solution to obtain a plasma-activated hyaluronic acid solution.

[0065] A plasma-activated hyaluronic acid solution was mixed with deionized water at a volume ratio of 3:2 to obtain a homogeneous solution. No cross-linking treatment was performed.

[0066] Comparative Example 2 A 1% (w / v) hyaluronic acid solution was prepared and subjected to plasma irradiation activation treatment using a plasma dielectric barrier discharge device. The plasma irradiation activation treatment was carried out in an ambient air atmosphere for 60 minutes. The active particles generated by the plasma were then loaded into the hyaluronic acid solution to obtain a plasma-activated hyaluronic acid solution.

[0067] Prepare a 10% (w / v) gelatin solution at a temperature of 40°C. Dilute the hydrochloric acid with physiological saline to achieve a pH of 3.5 to obtain the gel precursor solution.

[0068] The plasma-activated hyaluronic acid solution and the gel precursor solution were mixed thoroughly at a volume ratio of 3:2 to obtain a mixed solution. No cross-linking treatment was performed. The resulting product was continuously stored at 40°C.

[0069] Comparative Example 3 A 1% (w / v) hyaluronic acid solution was prepared and subjected to plasma irradiation activation treatment using a plasma dielectric barrier discharge device. The plasma irradiation activation treatment was carried out in an ambient air atmosphere for 60 minutes. The active particles generated by the plasma were then loaded into the hyaluronic acid solution to obtain a plasma-activated hyaluronic acid solution.

[0070] Prepare a 20% (w / v) gelatin solution and a 15% (w / v) polyvinyl alcohol solution, with the gelatin solution temperature controlled at 40℃. Mix the liquid gelatin solution and polyvinyl alcohol solution evenly at a volume ratio of 1:1. Dilute the hydrochloric acid with physiological saline to adjust the pH to 3.5 to obtain the gel precursor solution.

[0071] The plasma-activated hyaluronic acid solution and the gel precursor solution were mixed evenly at a volume ratio of 3:2 to obtain a mixed solution. Finally, the mixed solution was cooled at 4°C for 30 minutes without repeated freeze-thaw treatment to obtain a composite gel without freeze-thaw treatment.

[0072] Table 1 shows the detection results of Examples 1-8 and Comparative Examples 1-3. The detection methods include H2O2 and NO2. - ONOO - / O2 - Concentration detection.

[0073] For gels that were not lyophilized, the H2O2 concentration was detected using a hydrogen peroxide detection kit (Beyotime). The hydrogen peroxide detection reagent from the kit was thawed in an ice bath; after liquefying the sample at 37°C, 50 μL was added to three wells of a 96-well plate, and then 100 μL of hydrogen peroxide detection reagent was added to each well. After mixing, the plate was incubated at room temperature in the dark for 30 min, and the A560 absorbance of each well was measured using a microplate reader (Thermo, Varioskan Flash). The H2O2 concentration was calculated based on the measured absorbance and a standard curve. The NO2 concentration was detected using a nitric oxide detection kit (Beyotime). - The concentration of NO2 was determined. Grissin I and Grissin II reagents from the kit were thawed in an ice bath. After liquefying the sample at 37°C, 50 μL was added to three wells of a 96-well plate. Then, 50 μL each of Grissin I and Grissin II reagents were added to each well sequentially. After mixing, the plate was incubated at room temperature for 10 min. The A540 absorbance of each well was then measured using a microplate reader (Thermo, Varioskan Flash). NO2 was calculated based on the measured absorbance and the standard curve. - Concentration. ONOO was detected using 1-hydroxy-2,2,6,6-tetramethyl-4-piperidinone (Tempone-H, MCE). - / O2 - Concentration. Tempone-H was melted in an ice bath; 10 μL of 10 mM Tempone-H and 90 μL of the sample liquefied at 37°C were immediately drawn into a 25 μL capillary tube, and characteristic spectral lines were detected using an electron spin resonance spectrometer (Bruker EMX). The specific concentration was determined by double integration of the characteristic spectral lines.

[0074] The antibacterial effect after 3 days of storage was evaluated using the agar diffusion method. An equal volume of the sample was placed on the surface of an agar plate inoculated with the bacteria to be tested, and the size of the inhibition zone was observed after incubation.

[0075] For lyophilized composite gel products, a rehydration solution must be used to restore them to a gel, sol, or coatable state before testing. The amount of rehydration solution added should be based on restoring the initial volume of the composite gel before lyophilization or the preset test volume. Subsequent testing steps are the same as described above.

[0076] Table 1. Detection results of Examples 1-8 and Comparative Examples 1-3 As shown in Table 1, compared with Comparative Example 1, Examples 1-8 all showed improvement in the decay rate of active particles, indicating that combining the gel precursor solution with the plasma-activated polymer solution and combining it with cross-linking treatment can effectively improve the retention capacity of active particles in the system and slow down the decay rate of active particles. The difference between Example 2 and Comparative Example 2 is that Example 2 underwent cooling treatment to form a gelatin physical gel, while Comparative Example 2 did not undergo cooling cross-linking and was continuously stored in an environment of 40°C; the concentration of active particles in Comparative Example 2 decayed faster and the duration of the biological effect was shorter, indicating that cross-linking treatment is beneficial to forming a stable gel skeleton and improving the fixation and sustained-release capacity of active particles. The difference between Example 1 and Comparative Example 3 is that repeated freeze-thaw treatment was not performed, resulting in the lack of physical crystallization regions and molecular chain entanglement structures formed during the freeze-thaw process. The uniformity and density of the internal pore structure of the gel were poor, and the binding capacity of active particles was weak, thus leading to a decrease in the retention capacity of active particles. Example 4 shows a freeze-dried composite gel product obtained through freeze-drying. The initial concentration of active particles after rehydration was lower than before freeze-drying, indicating that some active particles may have been lost during freeze-drying and rehydration. However, compared to Comparative Example 1, which did not form a gel network, the rate of active particle attenuation was significantly lower, indicating that the freeze-dried composite gel product still has a certain capacity for storing active particles and is beneficial for improving the convenience of storage, transportation, and use. Some examples still showed antibacterial effects after 3 days of storage, indicating that an acidic gel precursor environment with a pH of 3-4 helps maintain the system's biological activity. Examples 5-8 illustrate the applicability of the method in these examples to different polymer solutions and different plasma activation methods.

[0077] The composite gel prepared in this embodiment can significantly extend the storage time of active particles, making it suitable for applications that rely on temperature-sensitive networks to achieve controlled release and regulate active particles with different lifespans, thus facilitating the expansion of application functions. The freeze-dried gel still retains the ability to store and extend the storage time of active particles, making it suitable for applications in high-permeability environments that require chronic, continuous, storable, or easily transportable conditions.

[0078] It should be understood that the specific embodiments described above are merely illustrative or explanatory of the principles of the invention and do not constitute a limitation thereof. Therefore, any modifications, equivalent substitutions, improvements, etc., made without departing from the spirit and scope of the invention should be included within the protection scope of the invention. Furthermore, the appended claims are intended to cover all variations and modifications falling within the scope and boundaries of the appended claims, or equivalent forms of such scope and boundaries.

[0079] The present invention has been described above with reference to embodiments thereof. However, these embodiments are merely illustrative and not intended to limit the scope of the invention. The scope of the invention is defined by the appended claims and their equivalents. Various substitutions and modifications can be made by those skilled in the art without departing from the scope of the invention, and all such substitutions and modifications should fall within the scope of the invention.

[0080] Although embodiments of the present invention have been described in detail, it should be understood that various changes, substitutions, and modifications can be made to the embodiments of the present invention without departing from the spirit and scope of the invention.

[0081] Obviously, the above embodiments are merely illustrative examples for clear explanation and are not intended to limit the implementation. Those skilled in the art will recognize that other variations or modifications can be made based on the above description. It is neither necessary nor possible to exhaustively list all possible implementations here. However, obvious variations or modifications derived therefrom are still within the scope of protection of this invention.

Claims

1. A method for storing plasma-active particles, characterized in that, include: A polymer solution is subjected to plasma activation treatment to obtain a plasma-activated polymer solution, wherein the plasma-activated polymer solution includes plasma-active particles; The gel precursor solution is added to the plasma-activated polymer solution according to a preset ratio and mixed to obtain a mixed solution. The gel precursor solution includes at least one of gelatin solution and polyvinyl alcohol solution, and the gel precursor solution is acidic. The mixed solution was subjected to cross-linking treatment to obtain a composite gel loaded with plasma-active particles.

2. The method for storing plasma active particles according to claim 1, characterized in that, The polymer in the polymer solution includes at least one of hyaluronic acid, sodium hyaluronate, alginate, chitosan, cellulose, cellulose derivatives, pectin, starch, and dextran.

3. The method for storing plasma active particles according to claim 1, characterized in that, The apparatus used for plasma activation treatment includes one or more of the following: plasma jet apparatus, dielectric barrier discharge apparatus, sliding arc discharge apparatus, and spark discharge apparatus. The plasma activation treatment methods include irradiation, air blowing, bubbling, and a combination of irradiation and air blowing. The plasma irradiation activation treatment is carried out in an ambient gas atmosphere, and the working gas introduced for the gas blowing or bubbling activation treatment includes at least one of air, nitrogen, oxygen, argon and helium. The working gas flow rate for the plasma blowing or bubbling activation treatment is 0.1 SLM to 5 SLM. The plasma activation treatment time is 5 min to 90 min.

4. The method for storing plasma active particles according to claim 1, characterized in that, The concentration of the gelatin solution is 5% to 40% (w / v), the temperature of the gelatin solution is 37°C to 50°C, and the concentration of the polyvinyl alcohol is 5% to 25% (w / v).

5. The method for storing plasma active particles according to claim 1, characterized in that, The pH value of the gel precursor solution is 3 to 4; when the gel precursor solution includes the gelatin solution and the polyvinyl alcohol solution, the volume ratio of the gelatin solution and the polyvinyl alcohol solution is 1:0.2 to 5.

6. The method for storing plasma active particles according to claim 5, characterized in that, The concentration of polymer in the plasma-activated polymer solution is 0.5% to 2% (w / v). The preset ratio is the volume ratio of the plasma-activated polymer solution to the gel precursor solution, and the preset ratio is 1 to 5:

1.

7. The method for storing plasma active particles according to claim 6, characterized in that, The volume ratio of the gelatin solution to the polyvinyl alcohol solution is 1:0.5 to 2; The preset ratio is 1 to 2:

1.

8. The method for storing plasma active particles according to claim 1, characterized in that, When the gel precursor solution comprises the gelatin solution and the polyvinyl alcohol solution, the crosslinking treatment includes a cooling treatment and / or a freeze-thaw treatment; When the gel precursor solution includes a gelatin solution, the crosslinking treatment includes a cooling treatment; When the gel precursor solution includes a polyvinyl alcohol solution, the crosslinking treatment includes a freeze-thaw process; The cooling process includes cooling the mixed solution to 25°C or below and maintaining it for 5 min to 90 min; The freeze-thaw treatment includes freezing the mixed solution at -20℃ to -80℃ for 8h to 16h and thawing it at 20℃ to 40℃ for 4h to 8h. The freeze-thaw treatment is repeated 1 to 5 times.

9. The method for storing plasma active particles according to claim 1, characterized in that, The method further includes: The composite gel is freeze-dried to obtain a freeze-dried composite gel product, which includes: pre-freezing the composite gel at -20℃ to -80℃, and freeze-drying the pre-frozen composite gel under vacuum conditions to obtain a freeze-dried composite gel product. The composite gel and / or the freeze-dried composite gel product shall be stored under low temperature, light-proof and sealed conditions.

10. A product loaded with plasma-active particles, characterized in that, The product includes a composite gel and / or a lyophilized composite gel product, wherein the composite gel is prepared using the method described in any one of claims 1-8, and the lyophilized composite gel product is prepared using the method described in claim 9.

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