Antimicrobial hydrocolloid composition and wound dressing comprising diatom-derived biosilica, and manufacturing methods thereof

Abstract

The present invention relates to an antimicrobial hydrocolloid composition comprising diatom-derived biosilica, polyvinyl alcohol, alginate, and polyphenol, a wound dressing, and a method for manufacturing the same. The antimicrobial hydrocolloid composition and wound dressing of the present invention not only possess excellent absorbency and the ability to maintain a moist environment, but also exhibit outstanding antimicrobial properties, offering the advantage of effectively preventing wound contamination and bacterial infection in humid environments. Specifically, by using diatom-derived biosilica and polyphenol together, the present invention offers excellent antimicrobial properties while possessing the ability to rapidly and effectively absorb exudate, thereby providing the advantage of effectively preventing contamination and bacterial infection caused by exudate absorption. Furthermore, by using polyvinyl alcohol and alginate together, the present invention offers excellent adhesiveness, allowing it to effectively protect the wound site against external contamination and infection. Additionally, it offers the advantage of minimizing bacterial infection in humid environments and inducing regenerative skin formation and the promotion of granular tissue formation. Moreover, the present invention does not exhibit cytotoxicity, ensuring safety and offering the advantage of safe and versatile application to wound sites.

Description

Technology Field

[0001] The present invention relates to an antimicrobial hydrocolloid composition and a wound dressing and a method for manufacturing the same. Specifically, the present invention relates to an antimicrobial hydrocolloid composition and a wound dressing comprising diatom-derived biosilica, and a method for manufacturing the same. Background Technology

[0002] A wound is a condition in which tissue continuity is disrupted due to bodily injury caused by external pressure, such as tearing of the skin surface or the formation of defects in specific areas of the skin or tissues. It is a comprehensive concept encompassing a pathological state in which tissues forming the internal or external surface of a living body—such as skin, muscles, nerve tissue, bone, soft tissue, internal organs, or vascular tissue—are fragmented or destroyed. Wounds are broadly classified into chronic and acute wounds. Healing periods and treatment methods vary depending on the severity, extent, size, and type of the injury, necessitating pharmaceuticals that facilitate smooth wound healing and ensure effective complete recovery.

[0003] Wound dressings are a representative pharmaceutical product used for the effective treatment of wounds. Wound dressings are a type of medical device that performs functions such as hemostasis, absorption of exudate, skin protection, and prevention of infection. They play a role in inducing rapid wound healing by more effectively managing the four major stages of wound progression: hemostasis, inflammation, proliferation, and maturation. Currently used wound dressings generally fall under the category of dressings whose primary function is to prevent the wound site from drying out by creating a moist environment.

[0004] Conventional research on wound dressings has primarily focused on creating a moist environment for exudates. Consequently, there have been limitations in ensuring sufficient exudate absorption and antimicrobial properties. In wound treatment, it is crucial to transport various wound healing factors from exudate to the wound site through rapid absorption. Furthermore, in the case of conventional wound dressings that lack antimicrobial properties, contamination of the dressing itself may occur due to absorbed exudate, raising concerns about wound contamination and secondary infection. Therefore, there is a need for research and development of wound dressings that exhibit excellent antimicrobial properties along with superior exudate absorption. Prior art literature

[0005] (Patent Document 0001) KR 10-1511405 B1 The problem to be solved

[0006] The main purpose of the present invention is to provide an antimicrobial hydrocolloid composition and a wound dressing that effectively prevent wound contamination and bacterial infection in a humid environment by having excellent absorbency and ability to maintain a moist environment, as well as excellent antimicrobial properties, in order to solve the aforementioned conventional problems. In addition, the main purpose of the present invention is to provide a method for manufacturing the antimicrobial hydrocolloid composition and the wound dressing.

[0007] Specifically, one objective of the present invention is to provide an antimicrobial hydrocolloid composition comprising diatom-derived biosilica, polyvinyl alcohol, alginate, and polyphenol.

[0008] In addition, one objective of the present invention is to provide an antibacterial wound dressing comprising the above-mentioned antibacterial hydrocolloid composition.

[0009] In addition, one objective of the present invention is to provide a method for preparing an antimicrobial hydrocolloid composition comprising: a) a step of preparing a first mixture by mixing polyvinyl alcohol and alginate, and a step of preparing a second mixture by mixing diatom-derived biosilica and polyphenol; and b) a step of preparing a third mixture by mixing the first mixture and the second mixture.

[0010] In addition, one objective of the present invention is to provide a method for manufacturing an antibacterial wound dressing comprising: a) a step of preparing a first mixture by mixing polyvinyl alcohol and alginate, and a step of preparing a second mixture by mixing diatom-derived biosilica and polyphenol; b) a step of preparing a third mixture by mixing the first mixture and the second mixture; c) a step of preparing a fourth mixture by mixing glycerin into the third mixture; and d) a step of removing the supernatant of the fourth mixture and obtaining a precipitate.

[0011] The purpose of the present invention is not limited to the description above and is provided for all cases in which appropriate effects can be obtained by utilizing the present invention. means of solving the problem

[0012] The inventors conducted research to manufacture an antimicrobial hydrocolloid composition and wound dressing that not only have excellent absorbency and the ability to maintain a moist environment, but also possess excellent antimicrobial properties to effectively prevent wound contamination and bacterial infection in a moist environment. As a result, it was discovered that when diatom-derived biosilica and polyphenol are used together, the diatom-derived biosilica exhibits excellent absorbency due to its large surface area and porosity, allowing for rapid and effective absorption of exudate, while the polyphenol acts as an excellent crosslinking agent and exhibits excellent antimicrobial properties, thereby providing excellent protection against contamination and bacterial infection caused by the absorption of exudate. Based on this, the present invention was completed.

[0013] Specifically, the present invention provides an antimicrobial hydrocolloid composition comprising diatom-derived biosilica, polyvinyl alcohol, alginate, and polyphenol.

[0014] In addition, the present invention provides an antimicrobial hydrocolloid composition comprising glycerin in addition to the antimicrobial hydrocolloid composition.

[0015] In addition, the present invention provides an antimicrobial hydrocolloid composition in which frustrule or diatomite is used as the diatom-derived bio-silica in the antimicrobial hydrocolloid composition.

[0016] In addition, the present invention relates to the antimicrobial hydrocolloid composition, wherein the polyphenol tannic acid, larch ( Larix The present invention provides an antimicrobial hydrocolloid composition using one or more selected from the group consisting of spp.) extracts and urushiol.

[0017] In addition, the present invention provides an antibacterial wound dressing comprising the above-described antibacterial hydrocolloid composition.

[0018] In addition, the present invention provides a method for preparing an antimicrobial hydrocolloid composition comprising: a) a step of preparing a first mixture by mixing polyvinyl alcohol and alginate, and a step of preparing a second mixture by mixing diatom-derived biosilica and polyphenol; and b) a step of preparing a third mixture by mixing the first mixture and the second mixture.

[0019] In addition, the present invention provides a method for preparing an antibacterial hydrocolloid composition, comprising the step of c) mixing glycerin into the third mixture.

[0020] In addition, the present invention provides a method for manufacturing an antibacterial wound dressing comprising: a) a step of preparing a first mixture by mixing polyvinyl alcohol and alginate, and a step of preparing a second mixture by mixing diatom-derived biosilica and polyphenol; b) a step of preparing a third mixture by mixing the first mixture and the second mixture; c) a step of preparing a fourth mixture by mixing glycerin into the third mixture; and d) a step of removing the supernatant of the fourth mixture and obtaining a precipitate.

[0021] In addition, the present invention provides a method for manufacturing an antibacterial wound dressing, comprising the step of e) freezing and then thawing the obtained precipitate to produce it in a hydrocolloid state.

[0022] The present invention will be described in more detail below.

[0024] In one specific embodiment, the present invention provides an antimicrobial hydrocolloid composition comprising diatom-derived biosilica, polyvinyl alcohol, alginate, and polyphenol.

[0025] In the present invention, the diatom is a type of microalgae known to be capable of growing through suspended growth or attached growth in freshwater or seawater where light, nutrients, and moisture are present. The diatom grows by forming trustles, which are porous biosilica, on its outer shell. The trustles are composed of amorphous SiO2 and exhibit a high-dimensional nanoporous structure due to their porosity. Frustles can be easily produced by culturing the diatoms, and their structure and / or size may vary depending on the species of diatoms, culture or growth conditions, etc. Even after the diatoms die, the trustles do not decompose but remain in the form of sediment, which is diatomite. From the above perspective, the diatom-derived biosilica particles of the present invention are not limited to the type of diatom, the method of extracting biosilica from diatoms, the type and form, structure, etc. of biosilica, and may be a concept that encompasses all biosilica derived from diatoms without special limitations.

[0026] Non-limiting examples of the above-mentioned diatom-derived biosilica include the genus Coconase ( Cocconeis ), genus Planotidium ( Planothidium ), Cymbela genus ( Cymbella ), genus Callonis ( Caloneis schroederi ), Nichia ( Nitzschia ), Yurakoseira genus ( Aulacoseira ), genus Nabicularis ( Navicula It may include biosilica derived from diatoms such as ). In one embodiment of the present invention, the diatom-derived biosilica is of the genus Euracocereus ( Aulacoseira It may be derived from ). Bio-silica derived from the genus Euracoceiras mentioned above is characterized by having a cylindrical porous structure, and thus can exhibit superior absorption.

[0027] In one embodiment of the present invention, the diatom-derived bio-silica may be frustrule or diatomite, and in a more specific embodiment, the diatom-derived bio-silica may be diatomite. Diatomite has the advantage of helping to rapidly and effectively absorb exudate from the wound site by exhibiting a large surface area and porosity.

[0028] In the present invention, the diatom-derived biosilica can exhibit nano-sized porosity and excellent water absorption. The diatom-derived biosilica of the present invention is inexpensive and does not require a separate nanoparticle manufacturing process, making it easy to use commercially; furthermore, it exhibits superior water absorption capacity and biocompatibility compared to synthetic silica particles.

[0029] In the present invention, the diatom-derived bio-silica may be surface-modified to be hydrophilic. As a specific example, the diatom-derived bio-silica may be surface-modified to be hydrophilic through an alkaline treatment method, for example, etching using sodium hydroxide (NaOH). As another specific example, the diatom-derived bio-silica may be surface-modified to be hydrophilic by a Piranha treatment method, which involves mixing sulfuric acid and hydrogen peroxide to remove organic matter and hydroxylate the surface. Alternatively, it may be surface-modified to be hydrophilic by a UV / O treatment method, such as generating ozone through UV to create -OH groups on the surface of the bio-material.

[0030] In the present invention, the diatom-derived biosilica may be in the form of particles. The average diameter of the particles is not particularly limited and may vary in size, for example, from 4 to 20 μm. The average diameter of the particles may refer to the average of the diameters of 100% or less, 99% or less, 98% or less, 97% or less, 96% or less, 95% or less, 94% or less, 93% or less, 92% or less, 91% or less, or 90% or less of the diatom-derived biosilica particles. More specifically, the diameter may be the value obtained by measuring the maximum diameter of a single particle. More specifically, the average diameter of the particles may be 4 μm or more, 5 μm or more, 6 μm or more, 7 μm or more, 8 μm or more, 9 μm or more, 10 μm or more, 12 μm or more, 14 μm or more, 16 μm or more, or 18 μm or more, and may be 20 μm or less, 18 μm or less, 16 μm or less, 14 μm or less, 12 μm or less, 10 μm or less, 9 μm or less, 8 μm or less, 7 μm or less, 6 μm or less, 5 μm or less, 4 μm or less, 3 μm or less, or 2 μm or less.

[0031] In the present invention, the polyvinyl alcohol (PVA) refers to a type of water-soluble polymer also generally called vinyl alcohol resin. In a specific embodiment of the present invention, a hydrocolloid composition and an adhesive wound dressing were prepared using polyvinyl alcohol with excellent adhesive properties. The adhesive wound dressing has the advantage of being able to provide more effective protection to the wound site by blocking the ingress of external contaminants to the wound site and being excellent at preventing secondary infection compared to the non-adhesive case.

[0032] In the present invention, the alginate refers to a type of polysaccharide acid of the cell wall of brown algae, also generally called alginic acid. In a specific embodiment of the present invention, a hydrocolloid composition and a wound dressing are prepared using alginate, and the hydrocolloid composition and wound dressing using alginate have the advantage of minimizing bacterial infection in a humid environment and promoting regenerative skin formation and granular tissue formation.

[0033] In the present invention, the polyphenol generally refers to a type of aromatic alcohol compound derived from plants. Although the specific type of compound used in the present invention is not limited, in order to more effectively exhibit the excellent antibacterial activity and the role as an effective crosslinking agent of the present invention, tannic acid, larch ( Larix One or more selected from the group consisting of extracts of spp.) and urushiol may be used. When the above polyphenol is used, it can form non-covalent bonds with biopolymers and form hydrogen bonds with other polymer materials to act as an effective crosslinking agent, and has the advantage of exhibiting excellent antibacterial properties.

[0034] In the present invention, the tannic acid is generally known to be a type of polyphenol biomolecule that produces an astringent taste and can be obtained from extracts of oak bark, persimmon, chestnut, acorn, etc.

[0035] In the present invention, the above larch ( Larixspp.) is generally known as a type of deciduous coniferous tree also called Japanese larch or Japanese cypress, and the above-mentioned larch extract includes the extract itself or any formulation of extract that can be formed using the extract, such as an extract obtained using an extraction solvent from various organs of the larch, e.g., bark, leaves, roots, seeds, fruits, flowers, etc., a diluted or concentrated extract, or a dried product obtained by drying the extract. In a specific embodiment of the present invention, the above-mentioned larch extract may be a larch bark extract. The above-mentioned larch bark extract is rich in hydroxyl groups, so it can act as an effective crosslinking agent and has the advantage of exhibiting excellent antibacterial effects.

[0036] In the present invention, the urushiol is generally of the Anacardiaceae family ( Anacardiaceae It is known as the sap of the poison ivy, a deciduous tree belonging to the family Urushiol. Urushiol has the advantage of exhibiting excellent antibacterial effects by damaging the cell membranes of bacteria.

[0038] An antimicrobial hydrocolloid composition comprising diatom-derived biosilica, polyvinyl alcohol, alginate, and polyphenol according to one embodiment of the present invention may additionally include glycerin to the above components.

[0039] In the present invention, the glycerin is also generally referred to as glycerol and has the characteristics of being highly viscous and hygroscopic. When the glycerin is additionally added and used in the present invention, there is an advantage of effectively preventing moisture loss in the composition.

[0040] In the hydrocolloid composition of the present invention, the content or mixing ratio of each component of the diatom-derived biosilica, polyvinyl alcohol, alginate, and polyphenol included therein can be varied and appropriately adjusted depending on the physical properties of the desired composition or the target site to be applied.

[0041] In a specific embodiment of the present invention, for the purpose of exhibiting superior physical properties of the present invention, for example, when the hydrocolloid composition is prepared and used as a wound dressing in the form of a film or patch, the hydrocolloid composition of the present invention may comprise 0.5 to 20 (w / v)% of diatom-derived biosilica, 0.5 to 20 (v / v)% of polyvinyl alcohol, 0.05 to 10 (v / v)% of alginate, and 0.1 to 15 (v / v)% of polyphenol, and more specifically, may comprise 1 to 10 (w / v)% of diatom-derived biosilica, 1 to 10 (v / v)% of polyvinyl alcohol, 0.1 to 5 (v / v)% of alginate, and 0.5 to 7 (v / v)% of polyphenol. It may be possible. In addition, glycerin may be additionally included in addition to the above components, and to exhibit a superior moisture loss prevention effect, 0.5 to 20 (v / v)% of glycerin may be included, and more specifically, 1 to 10 (v / v)% of glycerin may be included. In a specific embodiment of the present invention, when preparing the hydrocolloid composition of the present invention, a hydrocolloid composition was prepared with a mixing ratio of 4 (w / v)% diatom-derived biosilica, 5.45 (v / v)% polyvinyl alcohol, 0.55 (v / v)% alginate, and 1.27 (v / v)% polyphenol. In addition, in a specific embodiment of the present invention, 4.55 (v / v)% of glycerin was additionally mixed in addition to the above components. When a wound dressing is manufactured using a hydrocolloid composition prepared with the above mixing ratio, there is an advantage of having superior tensile strength and water absorption rate.

[0043] In another specific embodiment, the present invention,

[0044] a) a step of preparing a first mixture by mixing polyvinyl alcohol and alginate, and preparing a second mixture by mixing diatom-derived biosilica and polyphenol; and

[0045] b) a step of mixing the first mixture and the second mixture to prepare a third mixture; thereby providing a method for preparing an antimicrobial hydrocolloid composition.

[0046] In step a) above, the temperature and time conditions for mixing each component can be varied and appropriately adjusted depending on the specific type of component used, for example, the specific type of diatom-derived biosilica or the type of polyphenol, and can also be varied and appropriately adjusted depending on the physical properties of the desired hydrocolloid composition or wound dressing, the application site, etc. In a specific embodiment of the present invention, when diatomite is used as the diatom-derived biosilica and tannic acid is used as the polyphenol, a first mixture is prepared by mixing polyvinyl alcohol and alginate at approximately 90°C for approximately 10 minutes, and a second mixture of tannic acid and diatomite is added to the first mixture at regular intervals for approximately 20 minutes.

[0047] In step a) above, there is no particular order of precedence between the process of preparing the first mixture and the process of preparing the second mixture, and the first mixture may be prepared first and the second mixture may be prepared, or the second mixture may be prepared first and the first mixture may be prepared, or the first mixture and the second mixture may be prepared simultaneously in time.

[0048] In step a) above, the diatom-derived biosilica is first mixed with polyphenol, rather than with other components such as polyvinyl alcohol or alginate, to form a mixture, and then mixed with the polyvinyl alcohol and / or alginate. In this case, there is an advantage in that the clumping of the diatom-derived biosilica is effectively prevented.

[0049] After step b) above, step c) of mixing glycerin into the third mixture may be further included.

[0051] In another specific embodiment, the present invention,

[0052] a) a step of preparing a first mixture by mixing polyvinyl alcohol and alginate, and preparing a second mixture by mixing diatom-derived biosilica and polyphenol;

[0053] b) a step of preparing a third mixture by mixing the first mixture and the second mixture;

[0054] c) a step of preparing a fourth mixture by mixing glycerin into the third mixture; and

[0055] d) a step of removing the supernatant of the fourth mixture and obtaining a precipitate; thereby providing a method for manufacturing an antibacterial wound dressing.

[0056] In step d) above, the process of removing the supernatant of the fourth mixture can be performed by a method generally practiced in the relevant technical field or a similar field, and can be varied and appropriately adjusted according to the specific type, content, mixing ratio, etc. of each component. In a specific embodiment of the present invention, the process was performed by leaving the fourth mixture for about 30 minutes, and then removing the supernatant to obtain the precipitate once the supernatant and the precipitate are separated.

[0057] After step d) above, step e) of freezing and thawing the obtained precipitate to produce a hydrocolloid state may be further included. Step e) can be performed by methods generally practiced in the relevant technical field or similar fields, and can be varied and appropriately adjusted according to the specific type, content, mixing ratio, etc. of each component. In a specific embodiment of the present invention, an antibacterial wound dressing was prepared by spreading the obtained precipitate into a mold, freezing it for approximately 12 hours, and then repeating the process of thawing it for approximately 4 hours as one cycle, approximately 3 times.

[0059] Terms not otherwise defined in the present invention shall be interpreted as having the meanings commonly used in the relevant technical field. Additionally, the expression "or" as used in the present invention may be interpreted as a concept including "and" unless otherwise noted.

[0060] The scope of the present invention is not limited by the specific descriptions disclosed herein, and each description and embodiment disclosed herein may be applied to each other description and embodiment. That is, all possible combinations of the various elements disclosed herein are to be interpreted as falling within the scope of the present invention. Furthermore, a person skilled in the art may recognize or identify a number of equivalents to specific embodiments of the present invention through ordinary experimentation, and such equivalents are to be interpreted as falling within the scope of the present invention. Effects of the invention

[0061] The present invention relates to an antimicrobial hydrocolloid composition comprising diatom-derived biosilica, polyvinyl alcohol, alginate, and polyphenol, a wound dressing, and a method for manufacturing the same.

[0062] The present invention has the advantage of providing an antimicrobial hydrocolloid composition and wound dressing that not only have excellent absorbency and the ability to maintain a moist environment, but also possess excellent antimicrobial properties, thereby effectively preventing wound contamination and bacterial infection in a humid environment. Specifically, by using diatom-derived biosilica and polyphenol together, the present invention exhibits excellent absorbency due to the large surface area and porosity of the diatom-derived biosilica, allowing for rapid and effective absorption of exudate. At the same time, the polyphenol acts as an excellent crosslinking agent and exhibits excellent antimicrobial properties, providing the advantage of preventing contamination and bacterial infection caused by the absorption of exudate. In particular, it has the advantage of preventing contamination of the hydrocolloid composition or dressing itself by the absorbed exudate, thereby exhibiting excellent antimicrobial properties that prevent contamination and secondary infection of the wound site. In addition, the present invention has the advantage of being able to effectively protect the wound site against external contamination and infection by using polyvinyl alcohol, which provides excellent adhesion, and by using alginate, which minimizes bacterial infection in humid environments and promotes regeneration and granular tissue formation. Furthermore, the present invention has the advantage of being safe and usable in various ways on wound sites, as it does not exhibit cytotoxicity, thereby ensuring safety. Brief explanation of the drawing

[0063] FIG. 1 is a schematic diagram (Fig. 1a) and a photograph (Fig. 1b) schematically illustrating an antibacterial hydrocolloid composition and a method for manufacturing a wound dressing according to one embodiment of the present invention. FIG. 2 is a graph showing the results of measuring the tensile strength of a wound dressing according to one experimental example of the present invention. FIG. 2a shows all the results of measuring the tensile strength of Comparative Example 1 (PA), Comparative Example 2 (PAT), Comparative Example 3 (PAD), and Example 2 (PATD), and FIG. 2b shows the results of measuring the tensile strength of Comparative Example 1 (PA) and Comparative Example 2 (PAT), which are difficult to confirm on the graph of FIG. 2a due to their low tensile strength, as a separate graph. Figure 3 is a photograph of the results of SEM (Scanning Electron Microscope) image analysis of a wound dressing according to one experimental example of the present invention. Figure 4 is a graph showing the FT-IR analysis results of a wound dressing according to one experimental example of the present invention. Figure 4a shows the analysis results of Comparative Example 1 (PA), Figure 4b shows Comparative Example 2 (PAT), Figure 4c shows Comparative Example 3 (PAD), and Figure 4d shows the analysis results of Example 2 (PATD). Figure 5 is a graph showing the results of a cytotoxicity test of a wound dressing according to one experimental example of the present invention. FIG. 6 shows E. coli of a wound dressing according to one experimental example of the present invention ( Escherichia coli )(Fig. 6a) and Staphylococcus aureus ( Staphylococcus aureus This is a photograph showing the results of the antibacterial activity experiment for (Fig. 6b). In the figure, A is distilled water; B, C, D, and E are sections treated with ampicillin (amp) 0.1 μg / mL, 1 μg / mL, 10 μg / mL, and 100 μg / mL, respectively. Specific details for implementing the invention

[0064] The present invention will be described in more detail below through specific embodiments. However, these embodiments are merely examples for explaining the present invention and should not be interpreted as limiting the scope of the present invention in any way.

[0066] [Example]

[0067] Example 1: Preparation of an antimicrobial wound dressing

[0068] An antimicrobial wound dressing was prepared using diatom-derived biosilica, polyvinyl alcohol (PVA), alginate (Alg), and polyphenol. In this example, diatomite (DE) (Perma-Guard diatomaceous Earth, Fossil Shell Flour, USA) was used as the diatom-derived biosilica, and sigma-aldrich Mw. 146,000 - 186,000, 99+% hydrolyzed polyvinyl alcohol (PVA) was used. In addition, the above alginate (Alg) used was sigma-aldrich alginic acid sodium salt from brown algae, and the above polyphenol used was tannic acid (TA) (sigma-aldrich tannic acid, ACS reagent gallotannin, tannin).

[0069] Specifically, 30 mL of 10% polyvinyl alcohol and 10 mL of 3% alginate were mixed at 90°C for 10 minutes, and then a mixture of 7 mL of 10% tannic acid and 2 g of diatomaceous earth was added to the polyvinyl alcohol and alginate mixture at regular intervals while mixing for 20 minutes. Next, 5 mL of 50% glycerin was added and mixed to prevent moisture loss. Afterward, the mixture was left for 30 minutes to separate the supernatant and precipitate; the supernatant was then removed, the mixture was spread into a mold, and frozen for 12 hours to form a shape. Once the shape was formed, it was placed in a Petri dish and thawed for 4 hours. After completing a total of three freeze-thaw cycles, the surface moisture was removed, and the prepared wound dressing was stored in a sealed package to prevent deformation, thereby producing a hydrocolloid composition and a hydrocolloid wound dressing (Figs. 1a and 1b). The packaging film used in this practical example was 3M Tegaderm film.

[0071] Example 2: Preparation of antimicrobial wound dressing according to optimization of mixing ratio

[0072] In order to optimize the mixing ratio of polyvinyl alcohol, alginate, tannic acid, and diatomite in the preparation of the antibacterial wound dressing of Example 1 above, the tensile strength and water absorption rate of samples with different mixing ratios of each component were measured, and Scanning Electron Microscope (SEM) image analysis was performed.

[0073] Specifically, based on 5.45 (v / v)% polyvinyl alcohol, experiments were conducted by selecting alginate amounts of 0.27 (v / v)%, 0.55 (v / v)%, and 0.82 (v / v)%. At 0.27 (v / v)%, the water absorption rate decreased and precipitate formation was found to be unfavorable, while at 0.82 (v / v)%, the tensile strength was found to be weak. Accordingly, 0.55 (v / v)% was selected as the optimal value.

[0074] Experiments were conducted with tannic acid selected at 0.91(v / v)%, 1.27(v / v)%, and 1.82(v / v)%. At 0.91(v / v)%, the interaction reaction between the materials was insufficient, which was unfavorable for precipitate formation, while at 1.82(v / v)%, the bonding was strong, causing the materials to tend to separate from each other. Therefore, 1.27(v / v)% was selected as the optimal value.

[0075] For diatomite, the experiment was conducted with concentrations of 2(v / v)%, 4(v / v)%, and 6(v / v)%. At 2(v / v)%, the increase in absorption rate due to the addition of diatomite was weak, while at 6(v / v)%, it showed an unfavorable tendency for precipitate formation. Therefore, 4(v / v)% was selected as the optimal value.

[0076] Based on the above experimental results, polyvinyl alcohol 5.45 (v / v)%, alginate 0.55 (v / v)%, tannic acid 1.27 (v / v)%, and diatomaceous earth 4 (w / v)% were finally selected as the optimized mixing ratio. In addition, the mixing ratio of glycerin added to prevent moisture loss was finally selected as 4.55 (v / v)%.

[0078] [Comparative Example]

[0079] Comparative Examples 1 to 3: Preparation of wound dressings with different mixed components

[0080] Comparative Examples 1 to 3 were prepared by the same method as in Example 1 above, but with different mixing ratios of polyvinyl alcohol (PVA), alginate (Alg), tannic acid (TA), and / or diatomite (DE) as shown in Table 1 below.

[0081] ingredient Content Comparative Example 1 (PA) Comparative Example 2 (PAT) Comparative Example 3 (PAD) Example 2 (PATD) Polyvinyl alcohol (PVA) 5.45(v / v)% 5.45(v / v)% 5.45(v / v)% 5.45(v / v)% Alginate (Alg) 0.55(v / v)% 0.55(v / v)% 0.55(v / v)% 0.55(v / v)% Tannic acid (TA) - 1.27(v / v)% - 1.27(v / v)% Diatomite (DE) - - 4(w / v)% 4(w / v)% Glycerin (Gly) 4.55(v / v)% 4.55(v / v)% 4.55(v / v)% 4.55(v / v)%

[0083] [Experimental Example]

[0084] Experimental Example 1: Measurement of Tensile Strength of Antimicrobial Wound Dressing

[0085] The following experiment was performed to compare the tensile strength of the antibacterial wound dressing of the present invention, prepared with the optimized mixing ratio according to Example 2 above, with that of Comparative Examples 1 to 3.

[0086] Specifically, the wound dressings prepared in Example 2 and Comparative Examples 1 to 3 were each cut to dimensions of 2 cm X 4 cm X 0.1 cm, and then their tensile strength was measured. Tensile strength was measured and compared based on the value at the fracture point, with stress (the force resisting per unit area) and stroke (the degree of elongation), and the measured values ​​are shown in Figure 2 and Table 2 below.

[0087] division Stress (N / mm 2 ) Stroke (mm) Comparative Example 1 (PA) 0.0024 9.168 Comparative Example 2 (PAT) 0.0082 17.314 Comparative Example 3 (PAD) 0.2638 31.642 Example 2 (PATD) 0.3496 20.316

[0088] As a result of the experiment, it was confirmed that the values ​​shown in Table 2 above were obtained. Accordingly, the hydrocolloid wound dressing using diatom-derived biosilica of the present invention, prepared according to Example 2 (PATD), showed results of 145.67% stress and 122% stroke compared to Comparative Example 1 (PA), 4,163% stress and 17% stroke compared to Comparative Example 2 (PAT), and 33% stress and -36% stroke compared to Comparative Example 3 (PAD), confirming that it had the best overall tensile strength. In addition, considering that the tensile strength values ​​increased particularly in Comparative Example 3 (PAD) and Example 2 (PATD) containing diatomite, it was confirmed that diatomite contributes to the improvement of the physical properties of the wound dressing film.

[0090] Experimental Example 2: Measurement of Swelling Ratio and Porosity of Antimicrobial Wound Dressing

[0091] The following experiment was performed to compare the moisture absorption rate and porosity of the antibacterial wound dressing of the present invention, prepared with the optimized mixing ratio according to Example 2 above, with that of Comparative Examples 1 to 3.

[0092] Specifically, the wound dressings prepared in Example 2 and Comparative Examples 1 to 3 were each cut into pieces measuring 1 cm X 1 cm X 0.1 cm, and their respective weights were measured. Then, each was immersed in 10 mL of distilled water for 2 hours, and their respective weights were measured again to calculate the water absorption rate.

[0093] [Equation 1] Water absorption rate

[0094]

[0095] In the case of porosity, the wound dressings prepared in Example 2 and Comparative Examples 1 to 3 were each cut into pieces measuring 1 cm X 1 cm X 0.1 cm, and their respective weights were measured. After each was soaked in 10 mL of 95% ethanol for 1 hour, their respective weights were measured again to calculate the porosity.

[0096] [Equation 2] Porosity

[0097]

[0098] The results of the above water absorption rate and porosity experiments are shown in Table 3 below.

[0099] Comparative Example 1 (PA) Comparative Example 2 (PAT) Comparative Example 3 (PAD) Example 2 (PATD) Water absorption rate 87.7% 25.3% 107.09% 39.63% porous 64.2% 39.6% 130.33% 49.04%

[0100] As a result of the experiment, Example 2 (PATD) showed lower water absorption and porosity than Comparative Example 1 (PA) and Comparative Example 3 (PAD), but showed higher results of 1.57 times absorption and 1.24 times porosity compared to Comparative Example 2 (PAT) without diatomaceous earth, confirming that it has excellent absorption and porosity. In addition, considering that Comparative Example 3 (PAD) and Example 2 (PATD) containing diatomaceous earth showed increased absorption and porosity compared to cases without diatomaceous earth, it was confirmed that the addition of diatomaceous earth contributes to the improvement of the absorption and porosity of the wound dressing film.

[0102] Experimental Example 3: SEM (Scanning Electron Microscope) Image Analysis

[0103] SEM analysis was performed on the antibacterial wound dressing of the present invention prepared with the optimized mixing ratio according to Example 2 above and Comparative Examples 1 to 3.

[0104] Specifically, after coating wound dressings according to Comparative Example 1 (PA), Comparative Example 2 (PAT), Comparative Example 3 (PAD), and Example 2 (PATD), respectively, the surface was observed by photographing it at 500X magnification using an SEM (Fig. 3).

[0105] As a result of the experiment, it was confirmed that the surfaces of Comparative Example 1 (PA) and Comparative Example 2 (PAT) were aligned, and that diatomaceous earth was added to the surfaces of Comparative Example 3 (PAD) and Example 2 (PATD). In particular, it was confirmed that the dispersion of diatomaceous earth was uniform in Example 2 (PATD). Furthermore, it was confirmed that the added diatomaceous earth maintained a cylindrical porous shape, thus not losing its absorption rate and porosity characteristics. Additionally, regarding Example 2 (PATD), there is a problem where a dispersant must be added separately because dispersion does not occur well in the use of conventional bio-silica such as diatomaceous earth; however, in the case of Example 2 (PATD), it was confirmed that the material could be loaded onto the surface in a well-dispersed form without the addition of a separate dispersant. In addition, it was confirmed that Example 2 (PATD) maintained its structure without damage despite undergoing various processes, such as mixing and freeze-thaw cycles. This undamaged structure exerts a favorable effect on increasing the absorption rate, such as by increasing the surface area to maximize absorption. Furthermore, when compared to Comparative Example 3 (PAD), which does not contain tannic acid, Example 2 (PATD) appeared as a form in which the diatomite was embedded in polyvinyl alcohol and alginate, whereas in Example 2 (PATD), the diatomite was dispersed through the interaction between the diatomite and tannic acid, so it was confirmed to exist in a form in which the diatomite is attached to the surface of the polyvinyl alcohol (Fig. 3).

[0107] Experimental Example 4: FT-IR (Fourier-transform infrared spectroscopy) analysis

[0108] FT-IR analysis was performed on the antibacterial wound dressing of the present invention prepared with the optimized mixing ratio according to Example 2 above and Comparative Examples 1 to 3.

[0109] Specifically, wound dressings according to Comparative Example 1 (PA), Comparative Example 2 (PAT), Comparative Example 3 (PAD), and Example 2 (PATD) were each prepared in film form, and then 600-4000 cm -1 Each sample was analyzed within the range. The results of analyzing each peak are as shown in Figure 4 and the following description.

[0110] 1) Comparative Example 1 (PA): 660 cm -1 C=C stretching, 1080 cm -1 CO stretching, 2430 cm -1 C=C stretching, 2580 cm -1 SH stretching, 2660 cm -1 OH stretching, 2900 cm -1 CH stretching, 3150 cm -1 OH stretching, 3350 cm -1 OH stretching

[0111] 2) Comparative Example 2 (PAT): 660 cm -1 C=C stretching, 1080 cm -1 CO stretching, 2430 cm -1 2580 cm -1 SH stretching, 1875 CH stretching

[0112] 3) Comparative Example 3 (PAD): 660 cm -1 C=C stretching, 1080 cm -1 CO stretching, 2430 cm -1 2580 cm -1 SH stretching, 3215 cm -1 3300 cm -1 OH stretching 1875 cm -1 CH stretching

[0113] 4) Example 2 (PATD): 660 cm-1 C=C stretching, 1080 cm -1 CO stretching, 2430 cm -1 2580 cm -1 -H stretching, 3000 cm -1 3215 cm -1 OH stretching, 1875 cm -1 CH stretching

[0115] Experimental Example 5: Cytotoxicity Test

[0116] Cytotoxicity tests were performed on the antibacterial wound dressing of the present invention prepared with the optimized mixing ratio according to Example 2 above and Comparative Examples 1 to 3.

[0117] Specifically, to examine the biological activity of the wound dressings according to Comparative Example 1 (PA), Comparative Example 2 (PAT), Comparative Example 3 (PAD), and Example 2 (PATD), the cytotoxicity results of each film were confirmed using L-929 mouse fibroblasts.

[0118] First, L-929 mouse fibroblasts were cultured in a 96-well plate in a CO2 incubator at a temperature of 37°C for 48 hours. Each wound dressing according to Comparative Example 1 (PA), Comparative Example 2 (PAT), Comparative Example 3 (PAD), and Example 2 (PATD) was prepared with a diameter of 5 mm and then sterilized with UV-C (wavelength: 100-280 nm) for 30 minutes inside a biological safety cabinet (BSC). The sterilized samples were cultured for 72 hours at a temperature of 37°C with 0.4 mL of RPMI 1640 growth medium added per 0.02 g of sample.

[0119] Next, the medium present in the supernatant of the 96-well plate was removed, and 100 μL of each cultured sample extract was dispensed and incubated for 48 hours. After 48 hours, the plates were washed twice with 100 μL of PBS (phosphate-buffered saline), then treated with 80 μL of RPMI 1640 growth medium and 20 μL of MTS reagent, and the absorbance at 490 nm was measured after 3 hours.

[0120] Cell viability was calculated by using each wound dressing extract according to Comparative Example 1 (PA), Comparative Example 2 (PAT), Comparative Example 3 (PAD), and Example 2 (PATD) at a concentration of 0.2 g / mL, and it was determined that there was no cytotoxicity if the cell viability was 70 to 80% relative to the control group (RPMI 1640 growth medium).

[0121] As a result of the experiment, Comparative Example 1 (PA) and Comparative Example 3 (PAD) did not exhibit toxicity, but cytotoxicity was confirmed in Comparative Example 2 (PAT) (Fig. 5). This indicates that the cell viability increased in Example 2 (PATD), to which diatomite was added to Comparative Example 2 (PAT), which exhibited cytotoxicity, confirming that the addition of diatomite can have an effect in preventing cytotoxicity. When the above results are taken into account, it was confirmed that cell stability was ensured in the wound dressing of Example 2 (PATD) of the present invention.

[0123] Experimental Example 6: Antimicrobial Activity Experiment

[0124] Antibacterial activity experiments were performed on the antibacterial wound dressing of the present invention prepared with the optimized mixing ratio according to Example 2 above and Comparative Examples 1 to 3.

[0125] Specifically, five wound dressings according to Comparative Example 1 (PA), Comparative Example 2 (PAT), Comparative Example 3 (PAD), and Example 2 (PATD) were each punched to form disc shapes, and then UV-C (wavelength: 100-280 nm) sterilization was performed. Subsequently, 100 μg / mL, 10 μg / mL, 1 μg / mL, and 0.1 μg / mL of ampicillin (amp) and 1 μL of distilled water were dispensed into each of the five disc-shaped wound dressings, placed on a culture medium inoculated with bacteria, and incubated in a 37°C incubator for 16 hours for observation. Escherichia coli was used as the bacterium. Escherichia coli )(Fig. 6a) and Staphylococcus aureus ( Staphylococcus aureus Experiments were performed using )(Fig. 6b) respectively.

[0126] Experimental results, E. coli ( Escherichia coli ) and Staphylococcus aureus ( Staphylococcus aureus In all cases, under conditions of ampicillin (amp) 100 μg / mL and 10 μg / mL, a clear zone (bacteriostatic zone) of similar size of 0.9±0.1 cm was confirmed in all wound dressings according to Comparative Example 1 (PA), Comparative Example 2 (PAT), Comparative Example 3 (PAD), and Example 2 (PATD). In the case of ampicillin 1 μg / mL and 0.1 μg / mL and distilled water, a clear zone was confirmed only in Comparative Example 2 (PAT) and Example 2 (PATD), which had tannic acid added; thus, while Comparative Example 1 (PA) and Comparative Example 3 (PAD) were not confirmed to have antibacterial activity, Comparative Example 2 (PAT) and Example 2 (PATD) were confirmed to have excellent antibacterial activity (Fig. 6).

[0128] This specification omits detailed descriptions of matters that can be sufficiently recognized and inferred by those skilled in the art, and various modifications are possible within the scope of not altering the technical concept or essential configurations of the invention, in addition to the specific examples described herein. Accordingly, the invention may be implemented in a manner different from that specifically described and exemplified in this specification, and this is a matter that can be understood by those skilled in the art.

Claims

Claim 1 An antimicrobial hydrocolloid composition comprising 0.5 to 20 (w / v)% diatom-derived biosilica, 0.5 to 20 (v / v)% polyvinyl alcohol, 0.05 to 10 (v / v)% alginate, and 0.1 to 15 (v / v)% tannic acid. Claim 2 An antimicrobial hydrocolloid composition according to claim 1, further comprising glycerin. Claim 3 An antimicrobial hydrocolloid composition according to claim 1, wherein the diatom-derived bio-silica is frustrule or diatomite. Claim 4 delete Claim 5 An antibacterial wound dressing comprising the antibacterial hydrocolloid composition according to any one of claims 1 to 3. Claim 6 a) a step of preparing a first mixture by mixing 0.5 to 20 (v / v)% of polyvinyl alcohol and 0.05 to 10 (v / v)% of alginate, and a step of preparing a second mixture by mixing 0.5 to 20 (w / v)% of diatom-derived biosilica and 0.1 to 15 (v / v)% of tannic acid; and b) a step of preparing a third mixture by mixing the first mixture and the second mixture; comprising a method for preparing an antimicrobial hydrocolloid composition. Claim 7 A method for preparing an antimicrobial hydrocolloid composition according to claim 6, further comprising the step of mixing glycerin into the third mixture. Claim 8 A method for preparing an antimicrobial hydrocolloid composition according to claim 6, wherein the diatom-derived bio-silica is frustrule or diatomite. Claim 9 delete Claim 10 a) a step of preparing a first mixture by mixing 0.5 to 20 (v / v)% of polyvinyl alcohol and 0.05 to 10 (v / v)% of alginate, and a step of preparing a second mixture by mixing 0.5 to 20 (w / v)% of diatom-derived biosilica and 0.1 to 15 (v / v)% of tannic acid; b) a step of preparing a third mixture by mixing the first mixture and the second mixture; c) a step of preparing a fourth mixture by mixing glycerin into the third mixture; and d) a step of removing the supernatant of the fourth mixture and obtaining a precipitate; comprising a method for preparing an antibacterial wound dressing. Claim 11 A method for manufacturing an antibacterial wound dressing according to claim 10, further comprising the step of e) freezing and then thawing the obtained precipitate to produce it in a hydrocolloid state. Claim 12 A method for manufacturing an antimicrobial wound dressing according to claim 10, wherein the diatom-derived bio-silica is frustrule or diatomite. Claim 13 delete

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