Antibacterial wet bandage comprising bacteriophage

Abstract

The present invention relates to an antibacterial wet bandage and a production method therefor, the antibacterial wet bandage comprising: a polymer layer having biodegradable and biocompatible characteristics; and an antibacterial coating layer comprising a bacteriophage, wherein the polymer layer may further comprise copper and zinc nanoparticles having an antiviral effect. The antibacterial wet bandage of the present invention may exhibit excellent antibacterial activity by the stable survival of the bacteriophage and may exhibit excellent antiviral capacity by means of the metal nanoparticles and thus, when used as a wet bandage attachable to a wound site, may effectively prevent infection which may occur at the wound site.

Description

Antibacterial wet bandage containing bacteriophage

[0001] The present invention relates to a wet dressing band attached to a wound site for treatment.

[0002]

[0003] Disposable bandages are usually attached to minor wounds on the skin to prevent bacterial infection and protect the wound from external stimuli.

[0004] Research has shown that keeping wounds moist is more effective for wound healing than keeping them dry. This has led to the development of bandages that maintain a moist environment to prevent exudate from drying out or becoming dehydrated. Accordingly, various foam-type moist bandages are now available.

[0005] With global plastic usage reaching record highs, the resulting plastic waste is skyrocketing, accelerating environmental pollution. The widespread use of disposable bandages also contributes to plastic waste, leading to a growing need for bandages made from biodegradable plastic that are environmentally friendly while retaining the properties of traditional plastics.

[0006] Furthermore, while conventional wet bandages can maintain a moist environment around the wound, they fail to effectively prevent bacteria and viruses from penetrating the wound, resulting in minimal infection prevention. Therefore, there is a pressing need for technologies that can maintain a moist wound environment while effectively preventing bacterial and viral infection.

[0007]

[0008] The present invention is intended to solve the above-described conventional problems and to provide an antibacterial wet bandage including a bacteriophage that can effectively prevent contact with bacteria and viruses that can cause infection at a wound site.

[0009] In addition, the present invention aims to provide an antibacterial wet bandage that can exhibit excellent antiviral effects by including copper and zinc nanoparticles.

[0010] Another object of the present invention is to provide an antibacterial wet bandage comprising a biocompatible and biodegradable antibacterial polymer.

[0011]

[0012] A wet band having an antibacterial and antiviral effect according to one embodiment of the present invention may include an adhesive layer; a film-like polymer layer positioned on the adhesive layer; and a bacteriophage dispersed on one surface of the polymer layer.

[0013] In one embodiment, the polymer layer may include biodegradable properties and include at least one selected from the group consisting of polyvinyl alcohol (PVA), polylactic acid (PLA), polycaprolactone (PCL), polybutylene succinate (PBS), polyethylene terephthalate (PET), polybutylene terephthalate (PBT), polyethylene (PE), polypropylene (PP), polyvinyl chloride (PVC), polyamide (PA), and polyurethane (PU).

[0014] In one embodiment, the polymer layer may comprise polylactic acid (PLA) and polycaprolactone (PCL) in a weight ratio of 99:1 to 70:30.

[0015] In one embodiment, the bacteriophage may have a killing ability against one or more selected from the group consisting of Staphylococcus aureus, pathogenic Escherichia coli, Salmonella, and Shigella.

[0016] In one embodiment, the bacteriophage may belong to the family Myoviridae.

[0017] In one embodiment, the bacteriophage comprises 10 3 PFU / cm 2 10 inland 9 PFU / cm 2 It may be included in the concentration of .

[0018] In one embodiment, the polymer layer further comprises metal nanoparticles dispersed in a region that does not contact the bacteriophage; wherein the metal nanoparticles may comprise one or more of copper and zinc nanoparticles.

[0019] In one embodiment, the polymer layer may include at least one selected from the group consisting of cellulose, chitosan, starch, hydrogel, polyethylene terephthalate (PET), polypropylene (PP), polyethylene (PE), ethylene vinyl alcohol (EVOH), ethylene vinyl acetate (EVA), polystyrene (PS), polyvinylidene chloride (PVDC), polyvinyl chloride (PVC), and polyethylene naphthalate (PEN).

[0020] In one embodiment, the polymer layer may include a first film layer including a surface on which bacteriophages are dispersed, and a second film layer including a surface on which metal nanoparticles are dispersed.

[0021] In one embodiment, the metal nanoparticles may be dispersed on the other surface of the polymer layer.

[0022] In one embodiment, the polymer layer may comprise a porous three-dimensional network structure.

[0023] According to one embodiment, the thickness of the layer in which the bacteriophage is dispersed may be 1 μm to 50 μm.

[0024] In one embodiment, the layer in which the bacteriophage is dispersed further comprises a thickener, and the thickener may comprise at least one selected from the group consisting of sodium alginate, gelatin, gum arabic, carboxymethylcellulose, guar gum, and kappa carrageenan.

[0025] According to one embodiment, the layer in which the bacteriophage is dispersed further comprises a plasticizer, and the plasticizer may comprise at least one selected from the group consisting of glycerol, sorbitol, trehalose, fructose, sucrose, mannitol, propylene glycol, and polyethylene glycol.

[0026] In one embodiment, the layer in which the bacteriophage is dispersed may include the thickener and the plasticizer in a weight ratio of 10:1 to 1:10.

[0027] A method for manufacturing a wet bandage having antibacterial and antiviral effects according to another embodiment may include the steps of: preparing an adhesive layer; forming a polymer layer on one surface of the adhesive layer; and dispersing a bacteriophage on one surface of the polymer layer.

[0028] In one embodiment, the dispersion of the bacteriophage may be formed by casting or coating a solution containing the bacteriophage onto one surface of the polymer layer.

[0029] In one embodiment, the solution comprises 10 5 PFU / cm 2 10 inland 11 PFU / cm 2 It may contain bacteriophages.

[0030] In one embodiment, the step of forming the polymer layer may further include: laminating two different polymer layers; and, after the step of dispersing the bacteriophage, dispersing copper and zinc nanoparticles in an area of ​​the polymer layer that does not come into contact with the bacteriophage.

[0031] According to one embodiment, the step of dispersing bacteriophage on one surface of the polymer layer may further include the step of adding a thickener and a plasticizer to a solution containing bacteriophage.

[0032]

[0033] The antibacterial wet band of the present invention has an antibacterial and antiviral effect that can effectively prevent a wound site from being infected with bacteria and viruses by including a film layer containing bacteriophage or copper and zinc nanoparticles.

[0034] However, the effects of the present invention are not limited to the effects described above, and include all effects naturally implemented due to the various configurations proposed in the present invention.

[0035]

[0036] Figure 1 shows the results of measuring the moisture permeability according to the PLA / PCL ratio of a PLA / PCL film manufactured according to one embodiment of the present invention.

[0037] Figure 2 shows the results of measuring the moisture permeability according to the TBC content of a PLA / PCL / TBC film manufactured according to one embodiment of the present invention.

[0038] FIG. 3 shows a scanning electron microscope (SEM) image of a composite film manufactured according to one embodiment of the present invention.

[0039] Figure 4 shows the FT-IR analysis result spectrum of a composite film manufactured according to one embodiment of the present invention.

[0040] FIG. 5 illustrates the outer shape of a film according to the type of coating material according to one embodiment of the present invention.

[0041] Figure 6 shows the viscosity measurement results according to concentration of sodium alginate coating according to one embodiment of the present invention.

[0042] Figure 7 shows SEM images of sodium alginate coating at different concentrations according to one embodiment of the present invention.

[0043] Figure 8 shows AFM images of sodium alginate coating at different concentrations according to one embodiment of the present invention.

[0044] Figure 9 shows a transmission electron microscope (TEM) image of bacteriophage PBSA08 according to one embodiment of the present invention.

[0045] Figure 10 shows the results of measuring the antibacterial activity of bacteriophage PBSA08 by MOI according to one embodiment of the present invention.

[0046] Figure 11 shows the results of measuring the thermal stability of bacteriophage PBSA08 according to one embodiment of the present invention.

[0047] Figure 12 shows the results of measuring the pH stability of bacteriophage PBSA08 according to one embodiment of the present invention.

[0048] Figure 13 shows the results of analyzing the genome of bacteriophage PBSA08 according to one embodiment of the present invention.

[0049] Figure 14 shows the results of analyzing the lineage of bacteriophage PBSA08 according to one embodiment of the present invention.

[0050] Figure 15 shows the results of measuring the antibacterial activity of a biodegradable film manufactured according to one embodiment of the present invention.

[0051] Figure 16 is a graph showing the results of evaluating the cytotoxicity of copper and zinc nanoparticles according to one embodiment of the present invention.

[0052] Figure 17 shows an SEM image of the surface of a Cu / Zn-CNF polymer layer manufactured in one embodiment of the present invention.

[0053] Figure 18 shows the results of EDS analysis of a Cu / Zn-CNF polymer layer manufactured in one embodiment of the present invention.

[0054] Figures 19a and 19b show cross-sectional images of a CNF polymer layer (a) and a Cu / Zn-CNF composite polymer layer (b), respectively, manufactured in one embodiment of the present invention.

[0055] Figure 20 shows the results of measuring the water solubility of a Cu / Zn-CNF polymer layer manufactured in one embodiment of the present invention.

[0056] Figures 21a and 21b show the results of the antiviral effect analysis of nanoparticles used in one embodiment of the present invention against MS2 and MNV-1, respectively.

[0057] Figures 22a and 22b show the results of analysis of the antiviral effect of the Cu / Zn-CNF polymer layer manufactured in one embodiment of the present invention against MS2 and MNV-1 viruses, respectively.

[0058] Figures 23a to 23c show the experimental results of observing the antiviral persistence under frozen conditions (-18°C), refrigerated conditions (5°C), and room temperature conditions (25°C) for the Cu / Zn-CNF polymer layer manufactured in one embodiment of the present invention, respectively.

[0059] Figures 24a to 24c show the results of experiments in which antiviral persistence was observed under low humidity conditions (relative humidity 20-30%), medium humidity conditions (relative humidity 50-60%), and high humidity conditions (relative humidity 70-80%), respectively, for the Cu / Zn-CNF polymer layer manufactured in one embodiment of the present invention.

[0060] Figures 25a and 25b are graphs showing the results of measuring the MS2 antiviral activity according to the ratio of copper nanoparticles (a) and zinc nanoparticles (b) for the Cu / Zn-CNF polymer layer manufactured in one embodiment of the present invention, respectively.

[0061] Figures 26a and 26b are graphs showing the results of measuring the MNV-1 antiviral activity according to the ratio of copper nanoparticles (a) and zinc nanoparticles (b) for the Cu / Zn-CNF polymer layer manufactured in one embodiment of the present invention, respectively.

[0062] Figures 27a and 27b are graphs showing protein SDS-PAGE results of nanoparticle treatment groups for MS2 and MNV-1, respectively, in one embodiment of the present invention.

[0063] Figures 28a and 28b are graphs showing the results of calculating the band intensity of the nanoparticle treatment group for MS2 and MNV-1, respectively, in one embodiment of the present invention.

[0064] Figures 29a and 29b show the results of RT-qPCR analysis to elucidate the intracellular antiviral mechanism against MS2 and MNV-1, respectively, in one embodiment of the present invention.

[0065] Figures 30a and 30b are graphs showing the results of calculating RNA expression levels based on RT-qPCR analysis results for MS2 and MNV-1, respectively, in one embodiment of the present invention.

[0066] Figure 31 is a graph showing the results of measuring the antiviral activity against MS2 and MNV-1 of clams under freezing conditions (-18°C), refrigeration conditions (5°C), and room temperature conditions (25°C) using a Cu / Zn-CNF polymer layer manufactured in one embodiment of the present invention.

[0067]

[0068] The embodiments of the present invention are provided for the purpose of illustrating the technical concept of the present invention. The scope of the rights of the present invention is not limited to the embodiments presented below or the specific descriptions of these embodiments.

[0069] All technical and scientific terms used in this invention, unless otherwise defined, have the meanings commonly understood by those skilled in the art to which this invention pertains. All terms used in this invention have been selected for the purpose of more clearly explaining the invention and are not intended to limit the scope of the rights provided for in this invention.

[0070] Expressions such as “comprising,” “having,” and the like used in the present invention should be understood as open-ended terms that imply the possibility of including other embodiments, unless otherwise stated in the phrase or sentence in which the expression is included.

[0071] The singular expressions described in the present invention may include plural meanings unless otherwise stated, and this also applies to the singular expressions described in the claims.

[0072]

[0073] A wet band having an antibacterial and antiviral effect according to one embodiment of the present invention may include an adhesive layer; a film-like polymer layer positioned on the adhesive layer; and a bacteriophage dispersed on one surface of the polymer layer.

[0074] The above adhesive layer is a layer that allows the wet band to adhere to the skin, and may include any component that can adhere to the skin. In the present invention, the composition of the adhesive layer is not particularly limited.

[0075] Biodegradable polymer compounds are polymers that are broken down by naturally occurring microorganisms into low-molecular-weight substances, ultimately decomposing into water and natural byproducts such as carbon dioxide or methane gas. Representative examples include polylactic acid, polycaprolactone, and polyester. The utilization of these biodegradable polymers is expected to be a positive solution to the waste problem caused by the rapidly increasing use of disposable plastics.

[0076] In one embodiment, the polymer layer may include biodegradable properties and include at least one selected from the group consisting of polyvinyl alcohol (PVA), polylactic acid (PLA), polycaprolactone (PCL), polybutylene succinate (PBS), polyethylene terephthalate (PET), polybutylene terephthalate (PBT), polyethylene (PE), polypropylene (PP), polyvinyl chloride (PVC), polyamide (PA), and polyurethane (PU).

[0077] In one embodiment, the polymer layer may comprise polylactic acid (PLA) and polycaprolactone (PCL) in a weight ratio of 99:1 to 70:30.

[0078] In particular, the present invention can produce a biodegradable film exhibiting excellent mechanical properties and minimized phase separation by combining PLA and PCL. The weight ratio of the PLA and PCL may be 99:1 to 70:30, preferably 95:5 to 85:15. Within the above range, the combination of PLA and PCL can exhibit tensile strength and elongation suitable for application as a biodegradable film, and if it is outside the above range, phase separation may occur.

[0079] According to an embodiment of the present invention, it was confirmed that as the PCL content increases, the tensile strength decreases and the elongation increases, but when the PCL content is 20% or more, it was confirmed that the two materials are not well compatible and the phase separation phenomenon increases rapidly.

[0080] The polymer layer may include a biodegradable polymer and a compatibilizer.

[0081] The above compatibilizer can be used in conjunction with the biodegradable polymer to enhance compatibility between the biodegradable polymers, thereby simultaneously increasing tensile strength and elongation, thereby further improving mechanical properties. The compatibilizer can be selected from commercially available compatibilizers, but tributyl citrate (TBC) is most preferred.

[0082] The compatibilizer may be included in an amount of 0.1 wt% to 2 wt%, preferably 0.2 wt% to 1 wt%, and more preferably 0.3 wt% to 0.7 wt%, based on the total weight of the biodegradable polymer. If the compatibilizer is added in excessive amounts, the moisture permeability of the film may increase, which may result in problems such as improper film formation or deterioration of the physical properties of the film.

[0083] According to an embodiment of the present invention, the lowest moisture permeability was observed in the PLA / PCL film with 0.5 wt% of TBC added, and it was confirmed that when TBC was added more than that, the moisture permeability gradually increased, and at 2 wt%, the same high moisture permeability as that of the composite film without a compatibilizer added was observed.

[0084] The above polymer layer can exhibit high light blocking properties by including a biodegradable polymer and a compatibilizer, can exhibit high moisture content and excellent solubility, and can have a smooth surface that does not cause phase separation.

[0085]

[0086] The wet band of the present invention comprises bacteriophages dispersed on one surface of the polymer layer, and the bacteriophages can remain stable even after the wet band is manufactured, thereby exhibiting continuous antibacterial activity. Hereinafter, the layer in which the bacteriophages are dispersed is referred to as an antibacterial coating layer.

[0087] The term "bacteriophage" of the present invention refers to a virus that uses bacteria as its host, and may be abbreviated as "phage." Bacteriophages exhibit the activity of killing host bacteria through a lytic cycle and / or a lysogenic cycle. For example, according to a lytic cycle, a bacteriophage can infect a bacteria, multiply inside the cell, and then be released by destroying the cell wall of the host bacteria after multiplication, thereby killing the bacteria. Since one type of bacteriophage has killing ability only against a specific category of host bacteria, it is important to select an appropriate bacteriophage depending on the type of bacteria to be killed.

[0088] In one embodiment, the bacteriophage may have a killing ability against one or more selected from the group consisting of Staphylococcus aureus, pathogenic Escherichia coli, Salmonella, and Shigella.

[0089] The bacteriophage included in the present invention is not particularly limited as long as it is a bacteriophage that can exhibit an antibacterial effect against bacteria and viruses that can cause infection at a wound site, and it is preferable to use a bacteriophage that has excellent killing ability against Staphylococcus aureus.

[0090] In the present invention, the bacteriophage may preferably be bacteriophage PBSA08 (hereinafter referred to as "phage PBSA08"). The phage PBSA08 belongs to the Myoviridae family and has a genome size of 130,665 bp. Phage PBSA08 has excellent killing ability against Staphylococcus aureus, a food pathogen, and has high stability against heat and pH, so that an antibacterial coating layer containing or coated with the bacteriophage can be usefully applied to a wet bandage attached to a wound site.

[0091] The antibacterial wet band of the present invention can exhibit excellent antibacterial activity when used as a wet band by including an antibacterial coating layer containing such bacteriophage on a polymer layer.

[0092] In one embodiment, the bacteriophage comprises 10 3 PFU / cm 2 10 inland 9 PFU / cm 2 It may be included in a concentration of . Preferably 10 5 10 inland 7 PFU / cm 2 It can be included in a concentration range of . By including a bacteriophage in the above concentration range, the antibacterial wet band can exhibit effective antibacterial and antiviral effects.

[0093]

[0094] In one embodiment, the polymer layer further comprises metal nanoparticles dispersed in a region that does not contact the bacteriophage; wherein the metal nanoparticles may comprise one or more of copper and zinc nanoparticles.

[0095] The wet bandage of the present invention can exhibit antiviral effects due to the synergistic effect of copper and zinc nanoparticles. In particular, the composite film of the present invention exhibits excellent antiviral effects against viruses, including norovirus, and can maintain its antiviral activity under various temperature and humidity conditions. Furthermore, the nanoparticles exhibit no cytotoxicity and have low water solubility, making it suitable for use as a wet bandage applied to wounds.

[0096] In one embodiment, the polymer layer may include a biocompatible polymer, and may include at least one selected from the group consisting of cellulose, chitosan, starch, hydrogel, polyethylene terephthalate (PET), polypropylene (PP), polyethylene (PE), ethylene vinyl alcohol (EVOH), ethylene vinyl acetate (EVA), polystyrene (PS), polyvinylidene chloride (PVDC), polyvinyl chloride (PVC), and polyethylene naphthalate (PEN).

[0097] Preferably, the polymer layer may comprise cellulose, and the cellulose may be cellulose nanofibers (CNF). Cellulose nanofibers facilitate the acceptance of inorganic particles and exhibit excellent compatibility and bonding properties, particularly with copper and zinc nanoparticles. Furthermore, they are non-cytotoxic and exhibit excellent water resistance, making them suitable for use in wet bandages.

[0098] The content of the polymer included in the above polymer layer may be 0.01 wt% to 10 wt%, and preferably 0.1 wt% to 1 wt%, based on the total weight of the composition including the solvent. Accordingly, when an amount of metal nanoparticles capable of exhibiting antiviral activity is added in the present invention, a uniform film can be formed.

[0099]

[0100] The metal nanoparticles used in the present invention may include copper nanoparticles and zinc nanoparticles. Copper and zinc nanoparticles have relatively low toxicity and are easy to synthesize. When copper and zinc nanoparticles are used in a polymer layer, they exhibit excellent water resistance, making them suitable for use as a wet band.

[0101] In particular, the present invention can exhibit a high synergistic effect on virus inhibition even at low concentrations by using the copper and zinc nanoparticles in combination. Specifically, the present invention can exhibit both intracellular and extracellular antiviral mechanisms, targeting the norovirus extracellularly and binding to viral receptors to prevent intracellular entry, as well as inhibiting viral gene synthesis.

[0102] In this regard, in the embodiment of the present invention, it was confirmed that when copper nanoparticles and zinc nanoparticles were combined and used, they exhibited superior antiviral activity against norovirus compared to films containing each of them, and it was confirmed that the antiviral activity was continuously maintained under various temperature and humidity conditions.

[0103] In the present invention, the content of the copper nanoparticles may be 10 μM to 1,000 μM, preferably 50 μM to 500 μM, and the content of the zinc nanoparticles may be 1 mM to 200 mM, preferably 5 mM to 100 mM. The content of each nanoparticle is relative to the entire composition including the solvent, and can be adjusted within the above range so as to exhibit excellent antiviral properties against norovirus while preventing cytotoxicity by the nanoparticles.

[0104] In this regard, in an embodiment of the present invention, a significant inhibitory effect on norovirus was observed when copper nanoparticles were 50 μM or more and zinc nanoparticles were 5 mM or more, and it was confirmed that when these were combined and used at the minimum concentration, norovirus could be effectively inhibited compared to a film containing each nanoparticle.

[0105] The polymer layer may further include a plasticizer as a type of film base material. The plasticizer may be used in an amount of 20 to 60 wt%, preferably 30 to 50 wt%, based on the polymer weight.

[0106] The plasticizer may include at least one selected from the group consisting of glycerol, sorbitol, trehalose, fructose, sucrose, mannitol, propylene glycol, and polyethylene glycol.

[0107] In addition to the above components, the polymer layer may additionally include additives such as crosslinking aids, wetting agents, preservatives, and strength-giving agents as needed.

[0108] In the present invention, the composition forming the polymer layer may be in the form of a solution containing a solvent. The solvent may be appropriately selected and used depending on the base material used in film production. Preferably, water, saline solution, or the like may be used to produce a cellulose film.

[0109] In the present invention, by applying a combination of copper and zinc nanoparticles capable of producing a synergistic effect to a biocompatible polymer layer, an antibacterial wet bandage exhibiting a safe and excellent antiviral effect can be provided.

[0110]

[0111] In one embodiment, the polymer layer may include a first film layer including a surface on which bacteriophages are dispersed, and a second film layer including a surface on which metal nanoparticles are dispersed.

[0112] The polymer layer may be formed by laminating multiple layers, each including a layer containing dispersed bacteriophages and a layer containing dispersed metal nanoparticles. In this case, the polymer layer can simultaneously exhibit an antibacterial effect due to bacteriophages and an antiviral effect due to metal nanoparticles, thereby exhibiting an advantage in that it can exhibit a more powerful infection prevention effect.

[0113] For example, the layer in which the bacteriophage is dispersed may be a biodegradable film layer including PLA and PCL, and the layer in which the metal nanoparticles are dispersed may be a biocompatible film layer including cellulose nanofibers.

[0114] Since the antiviral effect of the above metal nanoparticles can affect bacteriophages, it is preferable that the metal nanoparticles and bacteriophages be dispersed in separate layers.

[0115] For example, the polymer layer may include a region where bacteriophages are dispersed and a region where metal nanoparticles are dispersed, and each region may be positioned on the same surface of the polymer layer. The polymer layer may have metal nanoparticles dispersed in a region excluding some regions where bacteriophages are dispersed.

[0116] In one embodiment, the metal nanoparticles may be dispersed on the other surface of the polymer layer.

[0117] For example, the polymer layer may include one surface on which bacteriophages are dispersed and the other surface on which metal nanoparticles are dispersed. In this case, the antibacterial effect of the bacteriophage and the antiviral effect of the metal nanoparticles can be effectively exerted without the bacteriophage and the metal nanoparticles being dispersed on the same surface.

[0118]

[0119] In one embodiment, the polymer layer may comprise a porous three-dimensional network structure.

[0120] The above polymer layer can have flexibility as it includes a network structure in which polymers are entangled with each other, and can be attached to the curved surface of the skin, thereby blocking the invasion of external infectious factors.

[0121] Since the mesh network structure of the polymer layer contains multiple pores internally, it can also have the effect of expanding the surface area into which bacteriophages or metal nanoparticles can be dispersed. Accordingly, a greater amount of bacteriophages or metal nanoparticles can be effectively dispersed and remain bound to the polymer layer, thereby exhibiting sustained antibacterial and antiviral effects.

[0122] The porous structure of the polymer layer indicates that the metal nanoparticles have excellent compatibility with the polymer matrix, particularly the cellulose matrix. In the porous structure, the pore size may be 10 nm to 10 μm, preferably 500 nm to 1 μm. In such a structure, the polymer and nanoparticles in the polymer layer can exhibit excellent bonding and compatibility, while also exhibiting excellent water resistance.

[0123] When the polymer layer includes a plasticizer such as sodium alginate, the porous network structure of the polymer layer can retain moisture for a long period of time, thereby enabling the long-term survival of bacteriophages. Furthermore, since moisture is not easily discharged to the outside due to capillary action within the porous network structure, the bacteriophages can be stably maintained within the structure. Accordingly, the wound area of ​​the skin to which the wet bandage of the present invention is attached can maintain an appropriate moist environment, while creating an environment in which the antibacterial and antiviral effects of the bacteriophages and metal nanoparticles can be continuously exerted.

[0124] In one embodiment, the polymer layer may include a relief structure. For example, the polymer layer may include relief structures having a predetermined interval, and a thicker antibacterial coating layer may be formed in the relatively deep grooves between the relief structures. Accordingly, the grooves may allow more moisture and bacteriophage to be dispersed, allowing the bacteriophage to survive for a longer period of time than the bacteriophage dispersed on the relief structures, and thus the antibacterial effect may be exhibited for a longer period of time.

[0125]

[0126] In one embodiment, the layer in which the bacteriophage is dispersed further comprises a thickener, and the thickener may comprise at least one selected from the group consisting of sodium alginate, gelatin, gum arabic, carboxymethylcellulose, guar gum, and kappa carrageenan. The antibacterial coating layer in which the bacteriophage is dispersed may preferably comprise sodium alginate as the thickener.

[0127] To stably maintain bacteriophages on the surface of a polymer layer, a viscous natural material is required. Such a material must have an appropriate viscosity that allows it to stably adhere to the film for a long time without separating or flowing. In the present invention, sodium alginate was selected as such a natural coating material and a bacteriophage coating solution was prepared. Sodium alginate is mainly used in foods to increase the viscosity of foods, enhance emulsification stability, and improve the physical properties and texture of foods. It is used as an adhesive, gelling agent, emulsifier, and thickener in various types of sauces, and is therefore a safe substance for the human body.

[0128] In one embodiment of the present invention, as a result of a coating experiment on sodium alginate, gelatin, gum arabic, carboxymethyl cellulose, guar gum, and kappa carrageenan, it was confirmed that gelatin, gum arabic, and carboxymethyl cellulose had too low a viscosity and did not adhere to the film but flowed, and that guar gum and kappa carrageenan had high viscosity of the coating solution and separated from the surface of the film, whereas sodium alginate adhered to the surface of the film and was able to form a very excellent coating layer.

[0129]

[0130] The antibacterial coating layer in which bacteriophages are dispersed may include a plasticizer along with sodium alginate. The plasticizer may serve to enable the coating layer to retain moisture, thereby enabling the bacteriophages to survive for a long period of time.

[0131] According to one embodiment, the layer in which the bacteriophage is dispersed further comprises a plasticizer, and the plasticizer may comprise at least one selected from the group consisting of glycerol, sorbitol, trehalose, fructose, sucrose, mannitol, propylene glycol, and polyethylene glycol.

[0132] The above plasticizer may preferably be glycerol. Glycerol has excellent water retention capacity, and thus, compared to other plasticizers, it exhibits excellent antibacterial activity by increasing the survival rate of bacteriophages even after film formation, and its long-term stability is ensured, allowing for sustained antibacterial activity.

[0133] In one embodiment, the layer in which the bacteriophage is dispersed may include the thickener and the plasticizer in a weight ratio of 10:1 to 1:10. The thickener and the plasticizer are more preferably in a weight ratio of 5:1 to 1:5, and most preferably in a weight ratio of 2:1 to 1:2.

[0134] In order to manufacture the biodegradable film of the present invention, in addition to the above components, additives such as a crosslinking aid, a wetting agent, a preservative, and a strength-giving agent may be additionally included as needed.

[0135]

[0136] The present invention also relates to a method for manufacturing the above antibacterial wet band.

[0137] A method for manufacturing a wet bandage having antibacterial and antiviral effects according to another embodiment may include the steps of: preparing an adhesive layer; forming a polymer layer on one surface of the adhesive layer; and dispersing a bacteriophage on one surface of the polymer layer.

[0138] The above polymer, bacteriophage and commercializing agent have been described in detail above.

[0139] First, a biodegradable polymer and a compatibilizer are dissolved / dispersed in a solvent to prepare a polymer coating solution, which is then coated to form a polymer layer. At this time, the solvent may be at least one organic solvent selected from the group consisting of chloroform, dichloromethane, and ethyl acetate.

[0140] The above biodegradable polymer can be mixed in a solvent at 1 to 10 wt%, preferably 3 to 7 wt%.

[0141] Additionally, the commercializing agent may be included in an amount of 0.1 to 2 wt%, preferably 0.2 to 1 wt%, and more preferably 0.3 to 0.7 wt%, based on the weight of the total biodegradable polymer, as described above.

[0142] In the present invention, it is preferable that the coating of the polymer layer is performed to a thickness of 40 µm to 100 µm, preferably 60 µm to 80 µm.

[0143] After coating the polymer layer, the drying step can be performed by drying at room temperature for 12 to 24 hours, and then drying at 40 to 70°C for an additional 3 to 10 hours.

[0144]

[0145] After manufacturing the polymer layer, an antibacterial coating solution containing bacteriophage can be coated on the polymer layer and dried to disperse the bacteriophage on the polymer layer.

[0146] According to one embodiment, the step of dispersing bacteriophage on one surface of the polymer layer may further include the step of adding a thickener and a plasticizer to a solution containing bacteriophage.

[0147] At this time, the antibacterial coating solution can be prepared by dispersing sodium alginate in a solvent, and the solvent can be preferably water. The concentration of sodium alginate in the antibacterial coating solution can be 1% (w / v) to 10% (w / v), more preferably 2% (w / v) to 4% (w / v), and most preferably 2.5% (w / v) to 3.5% (w / v). Within the above concentration range, the coating can have an appropriate viscosity to form a coating layer of uniform quality.

[0148] The above antibacterial coating solution may further comprise a plasticizer, preferably glycerol. The concentration of the plasticizer in the antibacterial coating solution may be 1% (w / v) to 10% (w / v), more preferably 2% (w / v) to 4% (w / v), and most preferably 2.5% (w / v) to 3.5% (w / v).

[0149] By dispersing sodium alginate and a plasticizer in a solvent to prepare a coating solution, a bacteriophage can be loaded into the solution to prepare an antibacterial coating solution.

[0150]

[0151] In one embodiment, the dispersion of the bacteriophage may be formed by casting or coating a solution containing the bacteriophage onto one surface of the polymer layer.

[0152] The antibacterial coating solution containing the above bacteriophage can be used as long as it can form a uniform surface through a solution process, and is not particularly limited in the present invention.

[0153] In one embodiment, the solution comprises 10 5 PFU / cm 2 10 inland 11 PFU / cm 2 It may contain a bacteriophage. The bacteriophage may be 10 7 10 inland 9It is more desirable to include it as PFU / mL.

[0154] According to one embodiment, the thickness of the layer in which the bacteriophage is dispersed may be 1 μm to 50 μm.

[0155] The thickness of the coated antibacterial coating layer is preferably 1 µm to 30 µm, more preferably 5 µm to 20 µm, and most preferably 10 µm to 15 µm. Within the above thickness range, an environment suitable for the survival of bacteriophages can be maintained.

[0156] For the coated antibacterial coating layer, drying can be performed at room temperature for 2 to 10 hours, preferably 3 to 5 hours, to form an antibacterial coating layer in which bacteriophages are maintained.

[0157]

[0158] In one embodiment, the step of forming the polymer layer may further include: laminating two different polymer layers; and, after the step of dispersing the bacteriophage, dispersing copper and zinc nanoparticles in an area of ​​the polymer layer that does not come into contact with the bacteriophage.

[0159] The molar ratio of the copper and zinc nanoparticles may be 1:20 to 1:200, preferably 1:50 to 1:150, and more preferably 1:80 to 1:120. Within this range, the copper and zinc nanoparticles may exhibit a superior synergistic effect compared to when each nanoparticle is included alone. In this regard, in an embodiment of the present invention, it was confirmed that when copper and zinc nanoparticles were added at a molar ratio of 1:100, a synergistic effect occurred on the antiviral activity of the copper and zinc nanoparticles, resulting in an excellent inhibitory effect against norovirus.

[0160] The polymer layer can be prepared by adding copper nanoparticles and zinc nanoparticles to a solution containing a polymer. Specifically, the solution can be prepared by dispersing a film base polymer in a solvent at 40°C to 80°C for 10 to 60 minutes, after which nanoparticles can be added and dispersed to prepare a composition containing metal nanoparticles. The composition can be dispersed in the polymer layer to form a thin film, and then dried at room temperature to 70°C for 10 to 60 hours.

[0161]

[0162] The biodegradable antibacterial film manufactured according to the present invention can exhibit excellent antibacterial effects by containing bacteriophage. In particular, the biodegradable antibacterial film of the present invention forms a bacteriophage layer containing sodium alginate on the film surface, thereby allowing the bacteriophage to survive stably for a long period of time after film formation, thereby exhibiting excellent antibacterial activity.

[0163] The thickness of the antibacterial wet band according to one embodiment of the present invention may be 0.01 mm to 2 mm, preferably 0.04 mm to 1 mm. As described above, the wet band with bacteriophage and metal nanoparticles dispersed therein can exhibit excellent antibacterial and antiviral properties even with a flexible and thin thickness, and thus can be used as a medical band that can be attached to a wound site.

[0164]

[0165] <Example>

[0166] The present invention is described in more detail through the following examples. However, these examples are provided to illustrate some experimental methods and compositions for the purpose of illustratively explaining the present invention, and the scope of the present invention is not limited to these examples.

[0167]

[0168] Manufacturing Example 1: Manufacturing of PLA / PCL composite film

[0169] PLA / PCL composite films of different compositions were manufactured using solvent-casting technology.

[0170] PLA and PCL pellets were dried in a drying oven at 60°C and 50°C, respectively, to remove any remaining moisture. PLA / PCL were mixed at weight ratios of 100 / 0, 90 / 10, 80 / 20, 70 / 30, 60 / 40, and 0 / 100, respectively, and each sample was dissolved in chloroform at a mass ratio of 1:20 of sample to solvent.

[0171] The mixture was stirred at room temperature (RT) for 6 hours and poured into a glass petri dish (150 x 150 mm) lined with a Teflon sheet. After drying at room temperature for 18 hours, all films were dried in a drying oven at 50°C for an additional 6 hours to completely remove any remaining solvent.

[0172] After separating the film from the petri dish, it was stored in a constant temperature-humidity chamber at 25°C and 50% relative humidity (RH) for at least 24 hours before the experiment.

[0173]

[0174] Experimental Example 1: Analysis of Mechanical Properties and Barrier Properties of PLA / PCL Composite Films

[0175] 1-1. Mechanical properties analysis of PLA / PCL composite films

[0176] The mechanical properties of the PLA / PCL film manufactured in Manufacturing Example 1 above were analyzed.

[0177] First, the thickness of each film was measured at five random points using a clearance gauge 547-401 (Mitutoyo, Kawasaki, Japan) with an accuracy of 0.001 mm. The average of each value was used to calculate the mechanical properties, moisture barrier properties, and optical properties.

[0178] Tensile strength (TS) and elongation at break (EAB) were measured using a texture analyzer (UTA-500N, Yeonjin, Seoul, South Korea) and are listed in Table 1 below.

[0179]

[0180] PLA / PCL ratio (wt%)Tensile strength (MPa)Elongation (%)100 / 030.89 ± 0.2413.76 ± 0.5490 / 1019.86 ± 1.7420.94 ± 1.4580 / 2016.68 ± 0.3922.18 ± 2.6870 / 3018.05 ± 0.0633.08 ± 2.4560 / 407.05 ± 0.6814.32 ± 2.300 / 1000.84 ± 0.79116.96 ± 4.12

[0181]

[0182] As can be seen in the table above, as the PCL content increased, tensile strength decreased and elongation increased. However, when the PCL content exceeded 20 wt%, the two materials were not compatible, and phase separation rapidly increased.

[0183] From the above results, PLA / PCL (90 / 10 wt%) with the lowest proportion of PCL was selected as the composition of the final composite film.

[0184]

[0185]

[0186] 1-2. Analysis of barrier properties of PLA / PCL composite films

[0187] To analyze the barrier properties of the PLA / PCL composite film, water vapor permeability (WVP) was measured.

[0188] The weight-per-vitreous viscosity (WVP) of the composite film was determined gravimetrically using the ASTM E96 standard cup method. The film was cut into a 65 mm diameter round shape and sealed in a circular cup containing anhydrous calcium chloride. Each sample was pre-weighed and stored in a constant temperature and humidity chamber at 25°C and 50% RH. The final weight of the sample was measured after 24 hours, and the WVP value was calculated using the following formula, which is shown in Figure 1.

[0189]

[0190] WVP(g / m 2 .h.kPa)=(β) / (Atβ)

[0191]

[0192] The above β is the weight of the cup (g) after 24 hours, x is the average thickness of the film (mm), and A is the exposed area of ​​the film (m 2 ), t is the exposure time (h), and β is the partial difference in water vapor pressure across the two sides of the film (Pa). The WVP of each film was measured three times.

[0193] As shown in Fig. 1, it was confirmed that the moisture permeability of the composite film tends to increase as the content of PCL, which has a relatively high moisture permeability, increases, and in the PLA / PCL (90 / 10) film finally selected in the previous experimental example 1-1, it was confirmed that the moisture permeability was at a level that was not statistically significantly different from that of the PLA single film.

[0194]

[0195] Manufacturing Example 2: Manufacturing of PLA / PCL / TBC composite film

[0196] In order to improve the compatibility of the PLA / PCL composite film, a PLA / PCL / TBC composite film was manufactured in the same manner as in Manufacturing Example 1, except that tributyl citrate (TBC), a compatibilizer that helps the connection between the two materials through hydrogen bonding, was added to the composition of PLA / PCL (90 / 10) selected in Experimental Example 1-1 at 0.5, 1, 3, and 5 wt% based on the weight of PCL.

[0197]

[0198] Experimental Example 2. Analysis of Mechanical Properties and Barrier Properties of PLA / PCL / TBC Composite Films

[0199] 2-1. Mechanical Properties Analysis of PLA / PCL / TBC Composite Films

[0200] The tensile strength and elongation of the PLA / PCL / TBC composite film manufactured in the above manufacturing example 2 were measured and are shown in Table 2 below.

[0201]

[0202] TBC content (wt%) Tensile strength (MPa) Elongation (%) 0 19.86 ± 1.74 20.94 ± 1.45 5 21.93 ± 2.33 21.02 ± 1.54 10 24.31 ± 2.0 1 21.08 ± 1.6 1 1 5 27.18 ± 1.90 21.20 ± 1.03 20 30.09 ± 0.80 23.34 ± 1.43

[0203]

[0204] As a result, it was confirmed that tensile strength and elongation increased simultaneously as the TBC content increased, as shown in Table 2.

[0205]

[0206] 2-2. Analysis of barrier properties of PLA / PCL / TBC composite films

[0207] The WVP value of the PLA / PCL / TBC composite film was calculated and shown in Fig. 2.

[0208] As can be seen in Fig. 2, the lowest moisture permeability was observed in the 5% TBC addition group, and when TBC was added in an amount of 5% or more, the moisture permeability gradually increased, and when 20% TBC was added, the moisture permeability was statistically the same as that of the PLA / PCL film without TBC addition.

[0209] Accordingly, PLA / PCL (90 / 10) + 5 wt% TBC (based on PCL weight) was selected as the final film composition.

[0210]

[0211] Experimental Example 3: Color difference and light transmittance analysis of composite films

[0212] The color difference and light transmittance of the PLA / PCL film and PLA / PCL / TBC film manufactured in the above manufacturing examples 1 and 2 were analyzed in comparison with the PLA film and PCL film.

[0213] The chromaticity of the film was observed using a colorimeter (CR-400 Chroma Meter, Konica Minolta Sensing, Inc., Japan). The equipment was set to a C light source and a 2° observation angle, and each measurement was repeated three times.

[0214] Transmittance was measured at 280 nm and 600 nm using a spectrophotometer (SP-UV 300, Perkin Elmer, UK) after cutting the film sample into 15 Х 100 mm pieces and fixing it in a film holder.

[0215] The results of measuring the color difference and transmittance of each film are shown in Table 3 below.

[0216]

[0217] Film L*a*b*△ET 280 (%)T 660 (%)PLA92.17 ± 0.29-0.23± 0.034.57± 0.210.39± 0.2361.57± 0.8593.69± 0.31PCL92.97 ± 0.13-0.21 ±0.024.49± 0.210.54± 0.180.93± 0.064.92± 0.15PLA 90 / PCL 10 92.55 ± 0.19-0.28 ±0.044.51± 0.070.27± 0.0644.59± 0.2184.91± 0.14PLA 90 / PCL 10 / TBC592.73 ± 0.14-0.27 ± 0.024.51 ± 0.120.30 ± 0.1845.04 ± 0.4184.31 ± 0.21

[0218]

[0219] In the above table, it was confirmed that the color difference of the film slightly increased in brightness and red when PCL and TBC were added to PLA.

[0220] In addition, the results of the light transmittance analysis of the film showed that the transparency of the film slightly decreased when PCL and TBC were added to PLA [T 660 (%)]. In addition, the results of the transmittance analysis in the ultraviolet region showed that the addition of PCL and TBC showed higher light blocking properties compared to the PLA single film [T 280 (%)].

[0221]

[0222] Experimental Example 4: Analysis of moisture content and solubility of composite films

[0223] Moisture content and solubility were analyzed for the composite film.

[0224] Film samples (20 x 20 mm) were pre-weighed (W0) and dried in a constant temperature and humidity chamber at 25°C and 50% RH for 24 hours. The dried film (W1) was weighed, placed in a laboratory crucible with 10 mL of distilled water, and stored at 25°C for 24 hours. The insoluble film was re-dried at 50°C for 24 hours, and the final weight (W2) was measured, and the moisture content and water solubility were calculated according to the following equations.

[0225]

[0226] Moisture content (%) = (W0- W1) / W0X 100

[0227] Solubility (%) = (W1- W2) / W1X 100

[0228]

[0229] The above W0 is the initial weight of the film, W1 is the weight of the film after oven drying, and W2 is the final weight of the film. Each sample was measured three times.

[0230] As shown in Table 4, PLA compared to PLA and PCL single films 90 / PCL 10 Composite films and PLA 90 / PCL 10 / TBC5 film showed higher moisture content, but solubility was not statistically different in all experimental groups.

[0231]

[0232] Film moisture content (%)Solubility (%)PLA0.012 ± 0.0161.015 ± 0.038PCL0.021 ± 0.0201.018 ± 0.024PLA 90 / PCL 10 0.057 ± 0.0460.987 ± 0.045PLA 90 / PCL 10 / TBC50.053 ± 0.0151.000 ± 0.020

[0233] Experimental Example 5: Surface Characterization of Composite Films

[0234] Scanning electron microscopy (SEM) analysis was performed to determine the surface properties of the composite film.

[0235] To examine the microstructure of the film using a scanning electron microscope (JSM-7610F, JEOL Ltd., Japan), samples were placed on carbon tape and coated with Pt / Pd for 30 s using a sputter coater (208HR, Cressington Scientific Instruments Ltd, UK). All samples were then observed at an acceleration voltage of 5.0 kV and a magnification of 5000X, and the results are shown in Fig. 3.

[0236] As confirmed in Fig. 3, when PCL was added to PLA, a relatively rough surface and pores due to the viscosity difference between the two materials were observed. In addition, PCL particles that were not sufficiently mixed into the PLA matrix were observed on the surface, so PLA 90 / PCL 10 It was confirmed that the composite film exhibited a phase separation phenomenon.

[0237] However, the films with added TBC had a relatively smooth surface, reduced pores, and fewer PCL particles, indicating that TBC is better than PLA. 90 / PCL 10 It was confirmed that it increases compatibility by participating in the connection of composite films.

[0238]

[0239] Experimental Example 6: FT-IR Analysis of Composite Films

[0240] The results of FT-IR analysis on the composite film and confirmation of the spectrum are shown in Fig. 4.

[0241] The FT-IR spectrum of the film was determined by an FT-IR spectrometer (Cary 630, Agilent Technologies Inc., Santa Clara, CA, USA) equipped with a reduced total reflection device, with a wavelength range of 4000-650 cm -1 4 cm in the spectral region -1 64 scans were performed at a resolution of .

[0242] As can be confirmed in Fig. 4, changes in wavelength region and peak intensity were observed in the spectrum of the PLA / PCL composite film. Compared to the PLA single film, there was a shift in the peak position in the C=O stretching vibration region, indicating that the C=O group of PLA and the OH group of PCL interact through hydrogen bonding. In addition, the increase in peak intensity of the C=O group of the PLA / PCL composite film upon addition of TBC confirmed that TBC contributes to the connection between PLA and PCL through hydrogen bonding.

[0243]

[0244] Experimental Example 7: Confirmation of coating quality according to coating material of composite film.

[0245] The coating properties of composite films were investigated using six natural materials: gelatin, gum arabic, carboxymethyl cellulose, guar gum, kappa carrageenan, and sodium alginate.

[0246] Glycerol, which has a high moisture retention capacity, was used as a plasticizer, and 4% (w / v) gelatin, 4% (w / v) gum arabic, 2% (w / v) carboxymethylcellulose, 2% (w / v) guar gum, 2% (w / v) kappa carrageenan, and 2% (w / v) sodium alginate were dispersed in distilled water together with 3% (w / v) glycerol, and heated to 60°C or higher while stirring until the solids were completely dissolved. The coating solution thus obtained was sufficiently cooled and then applied to PLA / PCL / TBC films.

[0247] Using a bar coater (RD Specialties, Inc., NY, USA), the coating solution was dispensed onto the film surface in a 13.72 μm moisture film, and the coated film was dried at 25°C for 4 hours. All film samples were stored in a constant temperature and humidity chamber at 25°C and 50% RH until the experiment. The appearance of the manufactured film is shown in Fig. 5.

[0248] As confirmed in Fig. 5, 4% (w / v) gelatin, 4% (w / v) gum arabic, and 2% (w / v) carboxymethyl cellulose had a very low viscosity of the coating solution, so it was confirmed that it flowed without adhering to the film.

[0249] In addition, 2% (w / v) guar gum and 2% (w / v) kappa carrageenan showed a phenomenon of clumping on the film due to the high viscosity of the coating solution, and a phenomenon of separation from the surface was confirmed.

[0250] On the other hand, it was confirmed that 2% (w / v) sodium alginate adhered very well to the film and formed a coating layer.

[0251]

[0252] Experimental Example 8: Analysis of Rheological Properties of Sodium Alginate Coating Solution

[0253] In order to confirm the rheological properties of the sodium alginate (SA) coating solution selected in the previous experiment, the viscosity characteristics according to the concentration of sodium alginate were confirmed.

[0254] First, the viscosity was measured according to the standard test method (ISO 3219). After preparing sodium alginate coating solutions having concentrations of 2, 3, and 4% (w / v) respectively in the same manner as in Experimental Example 7, the viscosity of the coating solutions was measured using a rotational rheometer (ARES-G2, TA Instruments, New Castle, DE, USA) with a 40 mm parallel plate shape at a gap of 0.5 mm.

[0255] Afterwards, analysis was performed in the shear rate range of 1-3000 1 / s at 25 ℃, and the viscosity was calculated using the following formula and shown in Figure 6.

[0256]

[0257] Viscosity (Pa.s) = Shear stress (Pa) / Shear rate (1 / s)

[0258]

[0259] As confirmed in Fig. 6, sodium alginate was confirmed to exhibit dynamic properties of a non-Newtonian fluid suitable for coating and paint solutions at all test concentrations [2, 3 and 4% (w / w)].

[0260] Additionally, when tested over a shear rate range used in typical industrial roller coating and brushing operations, it was confirmed that 3% sodium alginate exhibited the most suitable viscosity and dynamic properties as a coating solution compared to 2% and 4% sodium alginate.

[0261]

[0262] Experimental Example 9: Confirmation of Surface Characteristics of Sodium Alginate-Coated Composite Films

[0263] To determine the surface properties of the sodium alginate (SA) coated composite film, scanning electron microscopy (SEM) analysis was performed on films coated with various concentrations of sodium alginate, and the results are shown in Fig. 7.

[0264] Cross-sections of the coated films were examined. The low viscosity of 2% sodium alginate resulted in an uneven coating layer and incomplete adhesion to the film. Furthermore, the high viscosity of 4% sodium alginate resulted in the solution clumping and lifting from the film. However, 3% sodium alginate exhibited the highest adhesion to the film and a consistent thickness.

[0265] Next, the roughness of the surface coated with sodium alginate was analyzed using atomic force microscopy.

[0266] To perform atomic force microscopy (AFM) analysis, all coating films were cut into 10 mm Х 10 mm thin slices using a small, sharp knife and scanned in tapping mode on an NX10 AFM (Park systems, Suwon, South Korea) as shown in Fig. 8.

[0267] As shown in Figure 8, surface roughness increased as the sodium alginate content increased. While 2% sodium alginate exhibited the lowest surface roughness, its low viscosity prevented the coating from sufficiently adhering to the surface. Furthermore, 4% sodium alginate exhibited high surface roughness due to clumping.

[0268] Accordingly, the content of sodium alginate for the final coating was selected as 3%.

[0269]

[0270] Manufacturing Example 3: Preparation of bacteriophage solution

[0271] Staphylococcus aureus ATCC 29213 was used as the host bacterial strain, and bacteriophage PBSA08 (BP-6008) was obtained from the Bacteriophage Bank of Hankuk University of Foreign Studies (Yongin, Korea). Staphylococcus aureus strains were cultured in tryptic soy broth (TSB, MB cell, Seoul, Korea) and tryptic soy agar (TSA, MB cell) at 37°C under aerobic conditions, while other strains were cultured in media optimized for 37°C.

[0272] For phage propagation, the purified phage solution was infected into Staphylococcus aureus cultured for 1.5 hours (early exponential phase) and incubated at 37°C under aerobic conditions for 4 hours. The culture was then centrifuged at 15,000 x g at 4°C for 5 minutes, and the lysate was filtered using a 0.45 μm syringe filter (Whatman™ PVDF) to remove any remaining bacterial cells. This process was repeated three times, increasing the medium volume from 3 mL to 300 mL.

[0273] To prepare high-titer phage stocks, phage lysates were centrifuged at 30,000 × g for 20 min at 4°C. The supernatant was removed, and phage particles were dissolved in phosphate-buffered saline (PBS). Final phage titers were enumerated using the double-layer agar method. High-titer phage stocks were stored at 4°C until further experiments.

[0274]

[0275] Experimental Example 10: Morphological Characteristics of Bacteriophage PBSA08

[0276] For morphological analysis of bacteriophage PBSA08, transmission electron microscope (TEM) images were obtained and are shown in Figure 9.

[0277] In one embodiment, the bacteriophage may belong to the family Myoviridae.

[0278] For TEM analysis, phage suspension (10 9PFU / mL) were deposited on carbon-coated copper grids and negatively stained with 2% (v / v) uranyl acetate (pH 4.5). Phage morphology was determined using a transmission electron microscope (Libra 120, Carl Zeiss, Oberkochen, Germany) with an accelerating voltage of 120 kV, and phage size was determined as the average of five independent measurements.

[0279] As confirmed in Fig. 9, bacteriophage PBSA08 had an icosahedral head and a rigid, contractile tail. The head diameter was 95 ± 3 nm (n = 5), and the tail length and width were 168 ± 6 nm (n = 5) and 21 ± 1 nm (n = 5), respectively. Based on the structural characteristics described above, it was determined that the bacteriophage belonged to the Myoviridae family of the Caudovirales order.

[0280]

[0281] Experimental Example 11: Analysis of antibacterial activity of bacteriophage PBSA08

[0282] The antibacterial activity of bacteriophage PBSA08 against Staphylococcus aureus was confirmed at various multiplicity of infection (MOI).

[0283] First, an exponentially growing culture of Staphylococcus aureus ATCC 29213 (4 Х 10 8 50 mL of TSB containing 100 CFU / mL was infected with 1 mL of phage solutions at various multiplicities of infection (MOI) of 0.001, 0.01, 0.1, and 1, and cultures treated with 1 mL of PBS were used as a control. The cultures were aerobically cultured at 37°C for 24 h, and the antibacterial activity was confirmed by measuring the absorbance (600 nm) of the cultures every hour using a spectrophotometer (SP-UV 300, Perkin Elmer, UK). Each experiment was performed three times.

[0284] As shown in Figure 10, the bacteriophage-treated experimental group effectively inhibited bacterial growth compared to the control group at all MOIs. Furthermore, it was confirmed that all bacteriophage-treated groups exhibited sustained bacterial inhibition activity for up to 24 hours.

[0285]

[0286] Experimental Example 12: Confirmation of the host infection range of bacteriophage PBSA08.

[0287] To determine the host infection range of bacteriophage PBSA08, spot analysis was performed and is shown in Table 5.

[0288] Briefly, the phage solution (5 Х 10 8 After inoculating 10-fold serial dilutions of 10 PFU / mL) onto the culture surface of the target strain, the lysis sensitivity was determined when plaques or inhibition zones were formed in the strain. Plaque-opening (EOP) efficiency was determined using Staphylococcus aureus ATCC 29213 as the host strain, and was calculated by dividing the output phage titer for the target strain by the input phage titer of the host strain. EOP was evaluated as +++ (1 ≤ EOP), ++ (0.5 ≤ EOP < 1), + (EOP < 0.5), and - (not susceptibility).

[0289] As shown in Table 5, the range of host infection was confirmed, and 100% (14 / 14) of Staphylococcus aureus treated with bacteriophage PBSA08 were infected. In contrast, other strains were not infected, confirming that bacteriophage PBSA08 has high host specificity for Staphylococcus aureus.

[0290]

[0291] [Correction pursuant to Rule 91, July 8, 2024]

[0292]

[0293] Experimental Example 13: Stability Confirmation of Bacteriophage PBSA08

[0294] To confirm the stability of bacteriophage PBSA08, heat and pH stability were confirmed.

[0295] First, to check the thermal stability, 990 μL of PBS and phage solution (5 Х 10 8 PFU / mL) was mixed and incubated at -18°C to 70°C for 30 minutes.

[0296] Additionally, to check the pH stability, 990 μL of pH buffer (1-9) and 10 μL of phage solution (5 Х 10 8 PFU / mL) were mixed and incubated at 25°C for 30 minutes. The remaining phage concentration was measured using the double-layer agar technique, and the above experiments were repeated three times.

[0297] Referring to Figure 11, it was confirmed that bacteriophage PBSA08 was relatively stable under conditions of -18°C to 37°C.

[0298] Additionally, referring to Figure 12, it was confirmed that bacteriophage PBSA08 survives stably in the neutral range (pH 6-8).

[0299] Accordingly, bacteriophage PBSA08 is expected to be useful as a biological antibacterial material for application to various food groups.

[0300]

[0301] Experimental Example 14: Genome Analysis and Phylogenetic Analysis of Bacteriophage PBSA08

[0302] Genome analysis and phylogenetic analysis of bacteriophage PBSA08 were performed as follows.

[0303] First, phage genomic DNA was extracted using a modified standard phenol-chloroform extraction method. Briefly, phage solutions were treated with DNase I and RNase I to remove any remaining bacterial DNA / RNA contaminants. The purified phages were treated with lysis buffer containing 0.5 M EDTA (pH 8.0), 0.5% SDS, and proteinase K and incubated at 65 °C for 15 minutes. The mixture was then treated with an equal volume of phenol and centrifuged at 5,000 rpm for 5 minutes at room temperature.

[0304] The supernatant was then transferred to a new tube and immediately treated with phenol-chloroform-isoamyl alcohol (25:24:1). Next, the final supernatant was treated with sodium acetate (pH 5.2) and ethanol to precipitate DNA.

[0305] Finally, the phage DNA was concentrated in TE buffer (pH 8.0) and stored at -18 °C for further use.

[0306] The ORF (open reading frames) of the phage genome obtained by the above method were analyzed using RAST (https: / rast.nmpdr.org / ) and FgenesV software (http: / linux1.softberry.com / ).

[0307] ORFs were annotated using NCBI's BLAST (http: / blast.ncbi.nlm.nih.gov / ) and InterProScan programs and processed using the Artemis program.

[0308] The genome map was generated using Genescene (DNAstar, Madison, WI), and the phylogenetic tree was constructed using MEGA 11 software based on the major tail protein. The genome sequence of phage PBSA08 has been deposited in the GenBank database under accession number OP856857.

[0309] The results of the genome analysis of PBSA08 are shown in Figure 13, and the results of the phylogenetic analysis are shown in Figure 14.

[0310] In Fig. 13, bacteriophage PBSA08 was confirmed to be a safe lytic bacteriophage as a biological antibacterial material, as it does not have lytic, toxic, or antibiotic resistance genes.

[0311] In addition, referring to Figure 14, it was confirmed that it is phylogenetically close to Staphylococcus aureus bacteriophage SA11 and Staphylococcus aureus bacteriophage MR003, and through this, it was confirmed that bacteriophage PBSA08 also belongs to Staphylococcus aureus bacteriophage.

[0312]

[0313] Manufacturing Example 4: Manufacturing of a bacteriophage-coated composite film

[0314] To manufacture an antibacterial film coated with bacteriophage, 1 / 10% (v / v) of the phage stock of Manufacturing Example 3 was loaded into the 3% SA / glycerol solution selected in Experimental Example 9, and coated on the PLA / PCL / TBC film selected in Experimental Example 2. The phage loading concentration in the 3% SA / glycerol solution was 10 8 PFU / mL, and the concentration on the final film surface is 10 6 PFU / cm 2 am.

[0315] All film samples were stored in a constant temperature and humidity chamber at 25 °C and 50% RH until the experiment.

[0316]

[0317] Experimental Example 15: Confirmation of the antibacterial activity of a bacteriophage-coated composite film.

[0318] The antibacterial effect of a film treated with a coating solution containing bacteriophage PBSA08 was confirmed by comparing it with a composite film coated with sodium alginate not containing bacteriophage and a composite film not coated with sodium alginate.

[0319] In vitro, phage-coated film samples with a diameter of 25 mm (10 6 PFU / film) was cut into circles and placed in a 12-well microplate.

[0320] Add 2 ml of TSB to each well and add 20 μL of Staphylococcus aureus (10 6 CFU) was inoculated. Subsequently, the microplate was cultured under shaking conditions (120 rpm) at 25°C, and wells without film were used as controls. Bacterial growth inhibition activity was confirmed by measuring the culture absorbance at 600 nm every hour for 24 hours using a spectrophotometer (VarioskanTM LUX, Thermo Fisher Scientific, USA), and the results are shown in Figure 15.

[0321] In Fig. 15, in the case of the control group that did not contain bacteriophage, the bacteria grew and the absorbance continuously increased, but in the case of the film coated with bacteriophage, it was confirmed that the bacteria were effectively suppressed and the absorbance was low for up to 24 hours.

[0322] In summary of the above experimental results, it was confirmed that the film treated with a coating solution containing bacteriophage had excellent phage survival and exhibited an excellent antibacterial effect against Staphylococcus aureus.

[0323]

[0324] Manufacturing Example 5: Manufacturing of Cu / Zn-CNF composite polymer layer using metal nanoparticles

[0325] A Cu / Zn-CNF composite polymer layer was prepared using a base polymer and copper and zinc nanoparticles of the polymer layer.

[0326] To prepare the polymer layer, 0.5 g of cellulose nanofiber (CNF) powder (solid content 0.5 wt%) and 0.2 g of glycerol (40 wt% of CNF) were added to 100 mL of distilled water and dispersed by stirring at 60 °C for 30 minutes. Then, 50 μM of copper nanoparticles and 5 mM of zinc nanoparticles were added to the CNF solution and evenly dispersed using an ultrasonicator at 600 W for 20 minutes. Afterwards, the solution was uniformly applied to the substrate using casting and dried in a drying oven at 55 °C for 24 hours to obtain a Cu / Zn-CNF composite polymer layer.

[0327] The polymer layer, after drying, was conditioned for 24 hours at 25°C and 50% RH after peeling and then used in the experiment.

[0328]

[0329] Experimental Example 16: Confirmation of Cytotoxicity of Metal Nanoparticles

[0330] Cytotoxicity was evaluated using the MTT assay for 50 μM copper nanoparticles and 5 mM zinc nanoparticles.

[0331] Specifically, 2 X 10 per well in a 96-well plate 4 MTT assay was performed on cells, and 5 mg / mL of MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide) reagent was used to stain living cells. Afterwards, the formed formazan crystals were dissolved using DMSO, and the absorbance was measured under an optical density of 560 nm, and the results are shown in Fig. 16.

[0332] As a result, compared to the control group (100%) that was not treated with anything, the cell viability of the group treated with 50 μM copper nanoparticles and 5 mM zinc nanoparticles together was 89.99%, confirming that it did not exhibit cytotoxicity.

[0333]

[0334] Experimental Example 17: Structural Analysis of Cu / Zn-CNF Composite Polymer Layer

[0335] Scanning electron microscopy (SEM) and energy dispersive spectroscopy (EDS) analyses were performed on the Cu / Zn-CNF composite polymer layer of the above manufacturing example. Figure 17 shows an image of the film surface, Figure 18 shows the EDS analysis results (red Cu, green Zn) of the film surface, and Figures 19a and 19b show cross-sectional images of the CNF polymer layer (a) and the Cu / Zn-CNF composite polymer layer (b), respectively.

[0336] The above analysis results confirmed that the Cu / Zn-CNF polymer layer had a porous structure with pore sizes ranging from approximately 500 nm to 1 μm. This porous morphology of the CNF indicates that the nanoparticles have excellent compatibility with the CNF polymer matrix. In addition, EDS analysis results confirmed that copper nanoparticles and zinc nanoparticles were evenly distributed on the surface of the polymer layer at a constant ratio (average weight percentages of 2.17% and 53.32%, respectively) within the CNF polymer.

[0337]

[0338] Experimental Example 18: Characterization of Cu / Zn-CNF composite polymer layer

[0339] The water solubility was measured for the Cu / Zn-CNF composite polymer layer of the above manufacturing example, and the same experiment was performed for the CNF single layer, the Cu-CNF polymer layer, and the Zn-CNF polymer layer.

[0340] Figure 20 shows the results of water solubility measurements. According to the experimental results, when Cu / Zn nanoparticles were added to the CNF polymer layer, water solubility was significantly reduced compared to when Cu nanoparticles or Zn nanoparticles were added alone. Accordingly, it was confirmed that the joint treatment of Cu and Zn nanoparticles had a positive effect on reducing water solubility.

[0341] Since it is important to have high water activity and low water solubility for use in a wet bandage that comes into direct contact with wound exudate, it was found that the Cu / Zn-CNF composite polymer layer of the present invention has excellent water resistance due to the combined use of Cu and Zn, and thus can be usefully applied to a wet bandage.

[0342]

[0343] Experimental Method: Method for Measuring Antiviral Effect

[0344] Experiments were conducted using MS2 virus, a surrogate of human norovirus (HuNoV), and murine norovirus (MNV-1) as target viruses.

[0345] For growth media, MS2 virus was cultured in LB broth with Escherichia coli C-3000 (ATCC 15597) and MNV-1 was cultured in a medium containing high-glucose Dulbecco's Modified Eagle Medium (DMEM) and Fetal Bovine Serum (FBS) with macrophage cell line RAW 264.7. Afterwards, plaque assay was performed for quantitative measurement of MS2 virus and MNV-1.

[0346] For plaque assay evaluation, for MS2 virus, titer was measured after performing overlay technique on LB soft agar (0.4%) and LB agar plate. For MNV-1, RAW 264.7 cells were seeded at 5 X 10 per well in a 12-well plate. 5 The cells were seeded and cultured for 24 hours in a 5% CO2 atmosphere. Afterwards, the monolayer cells were infected with 100 μL of virus suspension, and the plates were cultured at 37°C for 1 hour. After the cells were added 1 mL of 2X DMEM and 1.5% agarose, the plates were fixed with 4% formaldehyde and stained with crystal violet to measure the titer.

[0347]

[0348] Experimental Example 19: Analysis of the in vitro antiviral effect of nanoparticles.

[0349] For in vitro antiviral evaluation, 50 μM of selected copper nanoparticles and 5 mM of zinc nanoparticles were added to 100 mL of distilled water, and then MS2 and MNV-1 were added to 10 6 Each was inoculated with 100 PFU / mL. Afterwards, the mixture was mixed in a shaking incubation for 1 hour, and the virus titer was measured using a plaque assay. The titer measurement results for MS2 and MNV-1 are shown in Figures 21a and 21b, respectively.

[0350] When copper nanoparticles were used alone in the MS2 virus, a 0.15-log reduction was observed, and when zinc nanoparticles were used alone, a 0.17-log reduction was observed. However, when copper and zinc nanoparticles were used together, a 4.86-log reduction was observed within 20 minutes, demonstrating high antiviral efficacy.

[0351] When copper nanoparticles were used alone in the MNV-1 virus, a 2.5-log reduction was observed, and when zinc nanoparticles were used alone, a 0.35-log reduction was observed. However, when copper and zinc nanoparticles were used together, a 4.42-log reduction was observed within 60 minutes, demonstrating high antiviral efficacy.

[0352] Through the above results, it was confirmed that the combination of copper nanoparticles and zinc nanoparticles showed an excellent synergistic effect that significantly reduced viral infection.

[0353]

[0354] Experimental Example 20: Analysis of the antiviral effect within the polymer layer of nanoparticles.

[0355] According to the ISO 21702 test method, the antiviral activity of Cu / Zn-CNF polymer layers was analyzed against MS2 and MNV-1 viruses. For comparison, the same experiments were performed on the CNF layer, Cu-CNF polymer layer, and Zn-CNF polymer layer, and the experimental results for MS2 and MNV-1 viruses are shown in Figures 22a and 22b, respectively.

[0356] As a result of the antiviral efficacy analysis against the MS2 virus, when copper nanoparticles were treated alone on the CNF polymer layer, a 1.19-log reduction was observed, and when zinc nanoparticles were treated alone, a 1.05-log reduction was observed. However, the Cu / Zn-CNF polymer layer treated with copper and zinc nanoparticles together showed a high antiviral efficacy, showing a 4.86-log reduction within 20 minutes.

[0357] When copper nanoparticles were treated alone on the CNF polymer layer against the MNV-1 virus, a 0.48-log reduction was observed, and when zinc nanoparticles were treated alone, a 0.07-log reduction was observed. However, when copper and zinc nanoparticles were treated together on the Cu / Zn-CNF polymer layer, a 0.85-log reduction was observed within 60 minutes, demonstrating high antiviral efficacy.

[0358] Through this, it was confirmed that the combination of copper nanoparticles and zinc nanoparticles showed an excellent synergistic effect in significantly reducing viral infection in the polymer layer.

[0359]

[0360] Experimental Example 21: Analysis of the antiviral durability of the Cu / Zn-CNF polymer layer.

[0361] With regard to temperature conditions, the antiviral persistence was evaluated for up to 3 days under frozen conditions (-18°C), refrigerated conditions (5°C), and room temperature conditions (25°C), and the results are shown in Figures 23a to 23c, respectively.

[0362] Experimental results showed that the Cu / Zn-CNF polymer layer with added copper and zinc nanoparticles showed superior antiviral efficacy compared to the group treated with only the CNF layer from day 0 to day 3 for both MS2 virus and MNV-1.

[0363] In addition, with regard to humidity conditions, the antiviral persistence was evaluated for up to 3 days under low humidity conditions (relative humidity 20-30%), medium humidity conditions (relative humidity 50-60%), and high humidity conditions (relative humidity 70-80%), and the results are shown in Figures 24a to 24c, respectively.

[0364] As a result, it was confirmed that the Cu / Zn-CNF polymer layer with added copper and zinc nanoparticles according to the present invention exhibited superior antiviral efficacy compared to the CNF-only polymer layer treatment group from day 0 to day 3 for both MS2 virus and MNV-1.

[0365]

[0366] Experimental Example 22: Antiviral Analysis According to Cu and Zn Ratio

[0367] The antiviral activity against MS-2 and MNV-1 was measured using the method of Manufacturing Example 5, but the concentration of copper nanoparticles was adjusted to 0, 50, 100, 250, and 500 μM, and the concentration of zinc nanoparticles was adjusted to 0, 5, 10, 25, 50, and 100 mM, respectively.

[0368] Figures 25a and 25b are graphs showing the results of measuring antiviral activity according to the ratio of copper nanoparticles (a) and zinc nanoparticles (b) for MS2, respectively, and Figures 26a and 26b are graphs showing the results of measuring antiviral activity according to the ratio of copper nanoparticles (a) and zinc nanoparticles (b) for MNV-1, respectively.

[0369] Experimental results showed that a significant reduction in MS2 and MNV-1 viruses was observed when the concentration of copper nanoparticles was 50 μM, and in particular, no plaques were detected at concentrations of 250 μM or higher for MS2 and 100 μM or higher for MNV-1. In addition, in the case of zinc nanoparticles, a significant difference in MS2 and MNV-1 viruses was observed starting from a concentration of 5 mM, and this reduction was confirmed to persist up to 100 mM.

[0370] That is, it was found that copper nanoparticles and zinc nanoparticles exhibited excellent antiviral activity starting from a minimum concentration of 50 μM and 5 mM, respectively, compared to the untreated nanoparticle group.

[0371]

[0372] Experimental Example 23: Analysis of changes in viral structural proteins following treatment with Cu and Zn nanoparticles.

[0373] To confirm the changes in viral structural proteins due to the synergistic effect of copper and zinc nanoparticles, the protein structures of MS2 and MNV-1 were observed for the 50 μM copper nanoparticle treatment group, the 5 mM zinc nanoparticle treatment group, and the copper (50 μM) / zinc (5 mM) nanoparticle treatment group.

[0374] Figures 27a and 27b are graphs showing the protein SDS-PAGE results of the nanoparticle treatment groups for MS2 and MNV-1, respectively, and Figures 28a and 28b are graphs showing the results of calculating band intensities of the nanoparticle treatment groups for MS2 and MNV-1, respectively.

[0375] Experimental results showed that MS2 and MNV-1 showed clear bands at 44 kDa (A-protein, the major structural protein of MS2) and 58.9 kDa (VP1, the major capsid viral protein of MNV-1), respectively. However, after exposure to nanoparticles, the protein band patterns changed significantly. Accordingly, it was found that copper and zinc nanoparticles degrade viral structural proteins.

[0376] In particular, the group treated with copper and zinc nanoparticles together exhibited a significantly greater intensity reduction in both MS2 and MNV-1 compared to the groups treated with either nanoparticle individually, with a particularly significant decrease in intensity observed for MNV-1. Accordingly, the synergistic effect of copper and zinc nanoparticles on the degradation of MS2 and MNV-1 structural proteins was confirmed.

[0377]

[0378] Experimental Example 24: Analysis of the antiviral mechanism according to treatment with Cu and Zn nanoparticles.

[0379] To elucidate the intracellular antiviral mechanism by copper and zinc nanoparticles, RdRP expression of MS2 and MNV-1 was confirmed by RT-qPCR analysis using specific primers as shown in Table 6 below.

[0380]

[0381] Virus Gene Sequence (5'-3') Function MS2A-protein F: TGT GGA GAG ACA GGG CAC TG Capsid component R: CAG TTG TTG GCC ATA CGG ATTR dRPF: CTC TGA GAG CGG CTC TAT TGG T Virus replication R: GTT CCC TAC AAC GAG CCT AAA TTCMNV-1VP1F: ATG AGG ATG AGT GAT GGC GC Capsid component R: TTA TTG TTT GAG CAT TCG GC CR dRPF: GGA ATT CCA TAT GCT TCC CCG CCC CTC AGG CAC CTA Virus replication R: ATA AGA ATG CGG CCG CAT CCT CAT TCA CAA AGA CTG CTG A

[0382]

[0383] Figures 29a and 29b are graphs showing the results of measuring the Ct values ​​of MS2 (a) and MNV-1 (b) during RT-qPCR for each treatment group, and Figures 30a and 30b are graphs showing the results of calculating the RNA expression level for each treatment group based on the Ct values.

[0384] The experimental results showed that the expression of replication genes and capsid synthesis genes was suppressed in the nanoparticle-treated group, and in particular, the Ct value of RdRP was significantly high in both MS2 and MNV-1, indicating that the expression of genes involved in replication by NP was suppressed.

[0385] Accordingly, it was confirmed that RdRP was significantly inhibited by a synergistic antiviral effect when copper and zinc nanoparticles were treated together, thereby significantly affecting the replication process.

[0386]

[0387] Experimental Example 25: Analysis of the anti-norovirus effect of the Cu / Zn-CNF polymer layer.

[0388] Depending on whether the Cu / Zn-CNF polymer layer of Manufacturing Example 5 was treated, the antiviral activity of the shellfish against MS2 and MNV-1 was analyzed under frozen conditions (-18°C), refrigerated conditions (5°C), and room temperature conditions (25°C), and the results are shown in Fig. 31.

[0389] Experimental results showed that under frozen conditions, MS2 showed a survival rate of 66.82% within 12 hours, and MNV-1 showed a survival rate of 82.77% within 24 hours, demonstrating excellent antiviral activity compared to the CNF polymer layer. Similarly, under refrigerated conditions, MS2 showed a survival rate of 65.67% within 6 hours, and MNV-1 showed a survival rate of 78.62% within 12 hours. Comparing frozen and refrigerated conditions, refrigerated conditions showed a higher antiviral effect in a shorter period of time.

[0390] Accordingly, it was confirmed that when the film of the present invention was applied to seafood, excellent antiviral activity was exhibited under freezing and refrigeration conditions.

[0391]

[0392] The above-described experimental examples confirmed the antibacterial effect using bacteriophages and the antiviral effect using copper and zinc nanoparticles. Therefore, the wet bandage of the present invention, with bacteriophages and copper and zinc nanoparticles dispersed in the polymer layer, demonstrated the potential for effective antibacterial and antiviral effects when applied to a wound.

[0393]

[0394] The above description is merely an illustrative example of the technical idea of ​​the present invention, and those skilled in the art will appreciate that various modifications and variations can be made without departing from the essential characteristics of the present invention. Therefore, the embodiments disclosed in the present invention are intended to illustrate rather than limit the technical idea of ​​the present invention, and the scope of the technical idea of ​​the present invention is not limited by these embodiments. The scope of protection of the present invention should be interpreted by the following claims, and all technical ideas within a scope equivalent thereto should be interpreted as being included in the scope of the rights of the present invention.

Claims

1. Adhesive layer; a film-like polymer layer positioned on the adhesive layer; and A bacteriophage dispersed on one side of the polymer layer; A moist bandage with antibacterial and antiviral properties.

2. In paragraph 1, The above polymer layer comprises biodegradable properties, Containing at least one selected from the group consisting of polyvinyl alcohol (PVA), polylactic acid (PLA), polycaprolactone (PCL), polybutylene succinate (PBS), polyethylene terephthalate (PET), polybutylene terephthalate (PBT), polyethylene (PE), polypropylene (PP), polyvinyl chloride (PVC), polyamide (PA), and polyurethane (PU). A moist bandage with antibacterial and antiviral properties.

3. In paragraph 1, The above polymer layer comprises polylactic acid (PLA) and polycaprolactone (PCL) in a weight ratio of 99:1 to 70:

30. A moist bandage with antibacterial and antiviral properties.

4. In paragraph 1, The above bacteriophage has a killing ability against at least one selected from the group consisting of Staphylococcus aureus, pathogenic Escherichia coli, Salmonella, and Shigella. A moist bandage with antibacterial and antiviral properties.

5. In paragraph 1, The above bacteriophage belongs to the Myoviridae family. A moist bandage with antibacterial and antiviral properties.

6. In paragraph 1, The above bacteriophage is 10 3 PFU / cm 2 10 inland 9 PFU / cm 2 which is included in the concentration of A moist bandage with antibacterial and antiviral properties.

7. In paragraph 1, The polymer layer further comprises metal nanoparticles dispersed in an area that does not come into contact with the bacteriophage; The above metal nanoparticles include at least one of copper and zinc nanoparticles. A moist bandage with antibacterial and antiviral properties.

8. In paragraph 7, The polymer layer comprises at least one selected from the group consisting of cellulose, chitosan, starch, hydrogel, polyethylene terephthalate (PET), polypropylene (PP), polyethylene (PE), ethylene vinyl alcohol (EVOH), ethylene vinyl acetate (EVA), polystyrene (PS), polyvinylidene chloride (PVDC), polyvinyl chloride (PVC), and polyethylene naphthalate (PEN). A moist bandage with antibacterial and antiviral properties.

9. In paragraph 7, The polymer layer comprises a first film layer including a surface on which bacteriophages are dispersed, A second film layer comprising a surface on which metal nanoparticles are dispersed, A moist bandage with antibacterial and antiviral properties.

10. In paragraph 7, The above metal nanoparticles are dispersed on the other side of the polymer layer, A moist bandage with antibacterial and antiviral properties.

11. In paragraph 1, The polymer layer comprises a porous three-dimensional network structure, A moist bandage with antibacterial and antiviral properties.

12. In paragraph 1, The thickness of the layer in which the bacteriophage is dispersed is 1 ㎛ to 50 ㎛, A moist bandage with antibacterial and antiviral properties.

13. In paragraph 1, The layer in which the bacteriophage is dispersed further comprises a thickener, The thickener comprises at least one selected from the group consisting of sodium alginate, gelatin, gum arabic, carboxymethylcellulose, guar gum, and kappa carrageenan. A moist bandage with antibacterial and antiviral properties.

14. In paragraph 1, The layer in which the bacteriophage is dispersed further comprises a plasticizer, The plasticizer comprises at least one selected from the group consisting of glycerol, sorbitol, trehalose, fructose, sucrose, mannitol, propylene glycol, and polyethylene glycol. A moist bandage with antibacterial and antiviral properties.

15. In paragraph 13 or 14, The layer in which the bacteriophage is dispersed contains the thickener and the plasticizer in a weight ratio of 10:1 to 1:

10. A moist bandage with antibacterial and antiviral properties.

16. Step for preparing the adhesive layer; A step of forming a polymer layer on one side of the adhesive layer; A step of dispersing bacteriophage on one side of the polymer layer; comprising; A method for manufacturing a wet bandage having antibacterial and antiviral effects.

17. In paragraph 16, The dispersion of the above bacteriophage is formed by casting or coating a solution containing the bacteriophage on one side of the polymer layer. A method for manufacturing a wet bandage having antibacterial and antiviral effects.

18. In paragraph 17, The above solution is 10 5 PFU / cm 2 10 inland 11 PFU / cm 2 Containing bacteriophages, A method for manufacturing a wet bandage having antibacterial and antiviral effects.

19. In paragraph 16, The step of forming the polymer layer includes laminating two different polymer layers, After the step of dispersing the above bacteriophage, A step of dispersing copper and zinc nanoparticles in an area of ​​the polymer layer that does not come into contact with the bacteriophage; further comprising; A method for manufacturing a wet bandage having antibacterial and antiviral effects.

20. In paragraph 16, A step of dispersing bacteriophage on one side of the polymer layer; A step of adding a thickener and a plasticizer to a solution containing a bacteriophage; further comprising; A method for manufacturing a wet bandage having antibacterial and antiviral effects.

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