Agastache rugosa exosome composition for skin barrier enhancement and Anti-inflammation, and use thereof

Abstract

To provide Agastache rugosa exosome composition for skin barrier enhancement and anti-inflammation, and use thereof.SOLUTION: The present disclosure provides a composition for skin barrier enhancement and anti-inflammation, including an exosome isolated from Agastache rugosa as an active ingredient. Also provided is the use of an exosome for producing skin barrier-enhancing and anti-inflammatory compositions, where the exosome is isolated from Agastache rugosa.SELECTED DRAWING: Figure 4

Description

[Technical Field]

[0001] The present disclosure relates to an exosome composition and uses thereof, and in particular to a Kawamidori exosome composition for skin barrier strengthening and anti-inflammation and uses thereof. [Background technology]

[0002] Skin tissue is primarily composed of the epidermis, dermis, and subcutaneous tissue, providing the body's first barrier defense and preventing the penetration of harmful substances such as allergens, irritants, and microorganisms. The epidermis is located at the outermost layer of skin tissue and can be divided into the stratum corneum, stratum lucidum, stratum granulosum, stratum spinosum, stratum basale, and basement membrane based on the multilayer structure of keratinocytes with different levels of differentiation. The stratum corneum is primarily responsible for the skin's barrier function and is composed of densely arranged keratinocytes. The stratum corneum is rich in lipid matrix and functions to prevent water loss. When the skin barrier is damaged due to aging or other adverse factors (including disease and UV exposure), transepidermal water loss increases, weakening the skin's resistance function. Environmental allergens can easily penetrate the skin, triggering immune and inflammatory responses, leading to the development of problems such as atopic dermatitis, psoriasis, allergic diseases, rosacea-like dermatitis, and facial eczema. In particular, the inflammatory response exacerbates abnormalities in the skin barrier function, and thus the inflammatory response and the skin barrier function form a vicious cycle.

[0003] Therefore, the prior art certainly needs to be improved in terms of how to strengthen the skin barrier function, maintain the stability of skin health, and alleviate conditions caused by "damage to the skin barrier," such as itching due to dry skin, increased wrinkles, and redness and fever due to inflammation. Summary of the Invention [Means for solving the problem]

[0004] One embodiment of the present disclosure provides a composition for strengthening skin barrier and anti-inflammatory, comprising exosomes isolated from Atractylodes mori as an active ingredient.

[0005] In some embodiments, exosomes are isolated from the aerial parts of Atractylodes oryzae.

[0006] In some embodiments, the exosomes have a mean particle size of between 20 nanometers and 500 nanometers.

[0007] In some embodiments, the exosomes have a concentration of 1×10 6 particles / ml ~ 1 x 10 12 Contains particles / ml.

[0008] In some embodiments, the exosomal transmembrane protein comprises CD9 (cluster of differentiation 9).

[0009] Another embodiment of the present disclosure provides a use of exosomes isolated from Atractylodes mori for the manufacture of a skin barrier strengthening and anti-inflammatory composition.

[0010] In some embodiments, the exosomal transmembrane protein comprises CD9.

[0011] In some embodiments, the exosomes are produced by the steps of harvesting the aerial parts of Atractylodes oryzae, adding a phosphate buffer to the aerial parts of Atractylodes oryzae followed by grinding to obtain a crude extract, filtering the crude extract to obtain a crude filtrate, and separating the crude filtrate by tangential flow filtration to obtain exosomes.

[0012] In some embodiments, the composition is a cosmetic composition, a pharmaceutical composition, or a food composition.

[0013] In some embodiments, the exosomes have a concentration of 1×10 6 particles / ml ~ 1 x 10 12 Contains particles / ml. [Brief explanation of the drawings]

[0014] The various aspects of the present disclosure will be best understood when reading the following detailed description in conjunction with the accompanying drawings. It should be noted that, in accordance with standard industry practice, the various features may not be drawn to scale. In fact, the sizes of the various features may be arbitrarily increased or decreased for clarity of discussion. To more clearly and comprehend the above and other objects, features, advantages, and embodiments of the present disclosure, the following description of the accompanying drawings is provided: [Figure 1] 1 is a transmission electron microscope image of a green alder exosome according to some embodiments of the present disclosure. [Figure 2] 1A-1C are sizes and concentrations of green alder exosomes according to some embodiments of the present disclosure. [Figure 3A] 1 is a high performance liquid chromatography (HPLC) chromatogram of a warm water extract of Atractylodes morifolium according to some embodiments of the present disclosure. [Figure 3B] 1 is an HPLC chromatogram of Kawamidori exosomes according to some embodiments of the present disclosure. [Figure 4] 1 shows a test of the protective activity of Kawamidori exosomes on keratinocytes according to some embodiments of the present disclosure. Statistical comparisons between groups were performed using one-way ANOVA for the experimental results, with ###p<0.0001 compared with the control group, *p<0.05, and ****p<0.0001 compared with the hydrogen peroxide (HO) group. The results are expressed as mean±standard deviation (SD) (N=3). [Figure 5] 1 is a microscopic observation of the protection of keratinocytes by Kawamidori exosomes according to some embodiments of the present disclosure. Cell images were recorded at a magnification of 10x10. [Figure 6] FIG. 1 shows that co-treatment of different concentrations of Bombyx mori exosomes with LPS according to some embodiments of the present disclosure does not affect cell viability. [Figure 7]1 shows a test of the anti-inflammatory activity of Kawamidori exosomes on immune cells according to some embodiments of the present disclosure. Statistical comparisons between groups were performed using one-way ANOVA. ##p<0.0001 compared with the control group, **p<0.01, and ****p<0.0001 compared with the bacterial lipopolysaccharide (LPS) group. Results are expressed as mean ± SD (N=3). [Figure 8] 1 shows the skin irritation test results of Kawamidori exosomes according to some embodiments of the present disclosure. [Figure 9] 1 shows eye irritation test results of Kawamidori exosomes according to some embodiments of the present disclosure. DETAILED DESCRIPTION OF THE INVENTION

[0015] To provide a more detailed and complete description of the present disclosure, the following provides illustrative descriptions of embodiments and specific examples of the present disclosure, but these are not the only ways to implement or use the specific examples of the present disclosure. The examples disclosed below may be combined with or substituted for each other when beneficial, and one example may be added to another example without further description or explanation. In the following description, many specific details are described in detail to enable the reader to fully understand the following examples. However, the embodiments of the present disclosure may also be implemented without these specific details.

[0016] Additionally, spatially relative terms such as "below," "above," and the like are intended to facilitate the description of one element or feature relative to another element or feature in the drawings. These spatially relative terms are intended to encompass different orientations of the device during use or operation in addition to the orientation depicted in the drawings. The device may be further positioned (e.g., rotated 90 degrees or at other orientations) and the spatially relative descriptions used herein may be interpreted accordingly.

[0017] As used herein, "a," "an," and "the" can generally refer to one or more, unless the context clearly indicates otherwise. It will be further understood that, as used herein, the terms "comprise," "include," "have," and similar terms designate stated features, regions, integers, steps, operations, elements, and / or components, but do not exclude the above or additional one or more other features, regions, integers, steps, operations, elements, components, and / or groups thereof.

[0018] Furthermore, when a number or range of numbers is described by "about," "approximately," and the like, the term is intended to cover the number within a reasonable range taking into account variations inherent in manufacturing as understood by those skilled in the art. For example, the number or range of numbers is intended to cover inclusive of the recited number, such as within a reasonable range of + / - 10% of the recited number, based on known manufacturing tolerances associated with manufacturing features (having the characteristics associated with the number).

[0019] Exosomes are small membrane-enclosed vesicles secreted from cells, ranging in size from 30 to 150 nm, and have a double membrane structure composed of proteins, lipids, and nucleic acids. Exosomes are involved in intercellular communication by transporting specific proteins, nucleic acids, and low-molecular-weight metabolites. Because exosomes have relatively low immunogenicity, tumor affinity, and targeting of innate and adaptive immune cells, they have become a trend in the development of treatments and delivery systems for multiple diseases in recent years.

[0020] Theoretically, any cell can secrete exosomes, and it has been proven that animal and human stem cell exosomes have multiple functions, such as promoting tissue regeneration, anti-aging, and immunoregulation, and related research has gradually expanded from medical functions to fields such as medical beauty and skin care products. However, in terms of supervision and management and application in skin care products, there are no clear standards and norms for animal stem cell exosomes.

[0021] Plants also secrete exosomes, called plant-derived exosome-like nanoparticles (PELNs). These tiny nanoscale vesicles secreted by plant cells contain substances such as DNA, small RNA (sRNA), microRNA (miRNA), and proteins. Plant exosomes have the advantages of being naturally sourced, easily mass-produced, low immunogenicity, zoonotic pathogen-free, highly bioavailable, and possessing potential biological activity. In addition, they also serve as a skin-penetrating carrier tool, helping active substances penetrate the skin via skin-related glandular pathways (hair follicles, sebaceous glands, and sweat glands), intercellular spaces, and cell penetration. Therefore, they are considered an economical and practical production source, and at the same time, they are safer and more compliant than animal- or human-derived stem cell exosomes, providing an opportunity for the development of innovative raw materials.

[0022] As used herein, the term "Agastache rugose exosomes", also referred to herein as "Agastache rugose exosome-like nanoparticles" or "Agastache rugose-derived exosomes", refers to nano-sized vesicles secreted by cells into the extracellular space.

[0023] In some embodiments, the Chloris sativa exosomes may be isolated from Chloris sativa.

[0024] In some embodiments, the Kawamidori exosomes have a particle size of 20 nanometers to 500 nanometers, including 25 nanometers, 30 nanometers, 35 nanometers, 40 nanometers, 45 nanometers, 50 nanometers, 60 nanometers, 70 nanometers, 80 nanometers, 90 nanometers, 100 nanometers, 150 nanometers, 200 nanometers, 250 nanometers, 300 nanometers, 350 nanometers, 400 nanometers, 450 nanometers, or any value between any two of these values.

[0025] In some embodiments, the Kawamidori exosomes are present at a concentration of, for example, 3×10 6 Particles / ml, 5 x 10 6 Particles / ml, 7 x 10 6 Particles / ml, 9 x 10 6 Particles / ml, 1 x 10 7 Particles / ml, 3 x 10 7 Particles / ml, 5 x 10 7 Particles / ml, 7 x 10 7 Particles / ml, 9 x 10 7 Particles / ml, 1 x 10 8 Particles / ml, 3 x 10 8 Particles / ml, 5 x 10 8 Particles / ml, 7 x 10 8 Particles / ml, 9 x 10 8 Particles / ml, 1 x 10 9 Particles / ml, 5 x 10 9 Particles / ml, 1 x 10 10 Particles / ml, 5 x 10 10 Particles / ml, 1 x 10 11 Particles / ml, 5 x 10 11 particles / milliliter, or any value between any two of these values, such as 1 x 10 6 particles / ml ~ 1 x 10 12 in the particles / milliliter range.

[0026] In some embodiments, the Kawamidori exosomes are obtained by methods including, but not limited to, ultra-high speed centrifugation, sucrose gradient centrifugation, microfiltration, tangential flow filtration, polymer precipitation, antibody magnetic bead separation, or a combination thereof.

[0027] In some embodiments, the Atractylodes mori exosomes may be obtained by a method comprising the steps of: harvesting the aerial parts of Atractylodes mori; adding a phosphate buffer to the aerial parts of Atractylodes mori and then grinding to obtain a crude extract; filtering the crude extract to obtain a crude filtrate; and separating the crude filtrate by tangential flow filtration to obtain Atractylodes mori exosomes.

[0028] In some embodiments, Kawamidori exosomes are used as active ingredients to enhance the skin barrier and provide anti-inflammatory effects.

[0029] In some embodiments, the present disclosure provides compositions for skin barrier strengthening and anti-inflammatory.

[0030] In some embodiments, the composition may be a cosmetic composition.

[0031] In some embodiments, the cosmetic composition may include functional additives and ingredients commonly found in cosmetic compositions. The functional additives may include, but are not limited to, polypeptides, polysaccharides, water-soluble vitamins, oil-soluble vitamins, or combinations thereof. The cosmetic composition may further include, but is not limited to, oils, fats, humectants, lubricants, surfactants, organic or inorganic pigments, organic powders, UV absorbers, preservatives, bactericides, antioxidants, plant extracts, pH regulators, alcohol, colorants, fragrances, blood circulation promoters, cooling agents, antiperspirants, purified water, or combinations thereof.

[0032] In some embodiments, the composition may be a pharmaceutical composition.

[0033] In some embodiments, the pharmaceutical composition comprises a Kawamidori exosome active ingredient and is administered to an individual via oral or parenteral routes. In some specific examples of the present disclosure, Kawamidori exosome is formulated and administered to an individual in an oral dosage form selected from the group consisting of a solution, suspension, emulsion, powder, troche, pill, syrup, buccal tablet, tablet, chewable capsule, and capsule.

[0034] In some embodiments, the pharmaceutical composition comprises a Kawamidori exosome active ingredient and a pharmaceutically acceptable carrier, wherein the pharmaceutically acceptable carrier is water, alcohols, glycols, preserving agents, antioxidants, solvents, emulsifiers, suspending agents, decomposers, binding agents, excipients, stabilizing agents, chelating agents, diluents, gelling agents, preservatives, lubricants, absorption enhancers, active agents, humectants, odor absorbers, fragrances, pH adjusting agents, occlusive agents, or the like. Agents, emollients, thickeners, solubilizing agents, penetration enhancers, anti-irritants, colorants, propellants, surfactants, and other similar carriers or carriers applicable to the present invention.

[0035] In some embodiments, the composition may be a food composition.

[0036] In some embodiments, the food composition may be added to edible materials in the form of a food additive, for example, to produce a food product for human or animal consumption. Food compositions include, but are not limited to, general foods, health foods, beverages, dietary supplements, dairy products, or feed. In an example of an oral dosage form, the food composition may optionally further comprise a food-acceptable carrier, excipient, and / or additive. In other examples, the dosage form of the multiple probiotic composition may include, but is not limited to, a powder, a lozenge, a granule, a suppository, a microcapsule, an ampoule, a liquid spray, or a stopper.

[0037] Hereinafter, the composition comprising Kawamidori exosomes of the present disclosure will be described in more detail by listing several examples and experimental examples. However, these are merely for illustrative purposes and are not intended to limit the present disclosure, and the scope of protection of the present disclosure is based on that defined by the appended claims.

[0038] The present disclosure provides a composition for strengthening the skin barrier and preventing inflammation, comprising exosomes isolated from Eucalyptus japonica as an active ingredient. Eucalyptus japonica exosomes have a particle size of 20 to 500 nanometers and may be isolated from Eucalyptus japonica plants. The composition containing Eucalyptus japonica exosomes as an active ingredient has the function of strengthening the skin barrier and exerting an anti-inflammatory effect by suppressing the production of nitric oxide without causing side effects to the skin.

[0039] Example

[0040] Although the methods disclosed herein are described below using a series of operations or steps, the order in which these operations or steps are presented should not be construed as limiting the present disclosure. For example, some operations or steps may be performed in a different order and / or simultaneously with other steps. Also, not all illustrated operations, steps, and / or features need to be performed to implement embodiments of the present disclosure. Also, each operation or step described herein may include multiple sub-steps or actions.

[0041] For clarity, features known in the art, features and elements that are not necessary for understanding the principles being described are omitted.

[0042] Production Example 1: Method for producing Kawamidori exosomes

[0043] The above-ground parts of the Atractylodes japonica plant were removed, washed to remove surface dirt, rinsed with pure water, and then dried. Three times the weight of phosphate buffer was added and the plant was crushed in a juicer to obtain a crude extract. The crude extract was subjected to solid-liquid separation, and the liquid portion was passed through a 0.22 micrometer (μm) filter membrane to remove impurities, obtaining a crude filtrate. The crude filtrate was concentrated using tangential flow filtration (TFF) with a 100 kilodalton (kDa) filter membrane to isolate Atractylodes japonica exosomes, also known as Atractylodes japonica-derived exosomes (AGS-TFF). The appearance of the purified Atractylodes japonica exosomes was examined using an electron microscope, and the particle size and concentration of Atractylodes japonica exosomes were analyzed using a nanoparticle size analyzer.

[0044] Example 1: Characterization of Kawamidori exosomes

[0045] The size and properties of the Kawamidori exosomes obtained in Production Example 1 were analyzed using a transmission electron microscope (TEM). Figure 1 shows that the Kawamidori exosomes have a diameter of 20 to 500 nanometers and are approximately spherical. Using an Exoid Nanoparticle Analyzer, the concentration of Kawamidori exosomes per milliliter (mL) of unit volume was found to be 5.89 x 10 9 It was confirmed that there were 100 individuals (Figure 2).

[0046] Example 2: Analysis of the specific marker CD9 transmembrane protein of Kawamidori exosomes

[0047] Previous studies have reported that plants contain a transmembrane structure similar to animal exosome CD9. Therefore, in this example, we attempted to detect the marker using a CD9 detection kit (ExoELISA-ULTRA Complete Kit, Cat. No. EXEL-ULTRA-CD9-1, System Bioscience, UK). As shown in Table 1 below, the color reaction of the Kawamidori exosomes obtained in Preparation Example 1 can also be generated using this platform, compared with the color values ​​of the exosome standard. Therefore, in addition to the particle size and image of the Kawamidori exosomes obtained in Preparation Example 1 matching the characteristics of exosomes, it is inferred that the lipid bilayer membrane contains the CD9 transmembrane protein, a common exosome-specific marker.

[0048] [Table 1]

[0049] Example 3: HPLC analysis of Kawamidori exosomes

[0050] According to literature reports, the warm water extract of A. japonica generally contains two main metabolites: rosmarinic acid and tilianin. To compare the components of A. japonica extract and A. japonica exosomes, HPLC was selected to perform HPLC component analysis of these two samples. The HPLC analysis method is described as follows:

[0051] 1. Hot water extract: 10 g of the aerial parts of A. japonica were boiled in 100 mL of water for 1 hour, and the filtrate was concentrated under reduced pressure and then freeze-dried. 25 mg of the dried sample was dissolved in 1 mL of deionized water. The filtrate was passed through a 0.45 μm filter membrane and subjected to HPLC analysis.

[0052] 2. Kawamidori exosomes: Kawamidori exosomes prepared in Production Example 1 were passed through a 0.45 μm filter membrane and subjected to HPLC analysis.

[0053] HPLC analysis conditions: Mobile phase A: 0.1% formic acid / water Mobile phase B: 0.1% formic acid / acetonitrile [Table 2] Chromatographic column: ACQUITY UPLC® CSH™ C18 chromatography column (2.1*100 mm, 1.7 μm, Waters Corporation, Milford, MA, USA) Flow rate: 0.4mL / min, UV=330nm Column temperature: 35°C, sample injection volume: 2 μL

[0054] As a result, as shown in Figure 3A, the Kawamidori warm water extract certainly contained two components, rosmarinic acid and tilianin. However, as shown in Figure 3B, Kawamidori exosomes contained 5.89 x 10 9 At this concentration, two components, rosmarinic acid and tilianin, were not detected.

[0055] Example 4: Protective activity of skin keratinocytes

[0056] In this example, damage to human HaCaT keratinocytes was induced by simulating oxidative stress with hydrogen peroxide, and the protective activity against the cells was evaluated after adding Kawamidori exosomes obtained in Preparation Example 1. HaCaT keratinocytes were cultured in a DMEM culture medium containing 100 units / mL penicillin, 100 μg / mL streptomycin, and 10% fetal bovine serum in an environment of 5% CO2 and 37°C. First, 1 × 10 4 The cells were cultured in a 96-well culture plate for 24 hours, and the supernatant was removed the next day. 6 , 1.83×10 7 , 3.71 × 10 7 , 7.36 x 10 7 , 1.47×10 8 , 2.95×10 8 Cell culture medium containing Kawamidori exosomes (particles / mL) was added and incubated for 1 hour. Subsequently, H2O2 was added to a final concentration of 200 μM, and the cells were incubated in an incubator for 16 hours. On day 3, the cells were incubated with 0.5 mg / mL 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) for 1 hour, then lysed with dimethyl sulfoxide (DMSO). The cell viability of each group was evaluated by OD 570 The absorbance values ​​were expressed as a percentage after comparison with the control cell group.

[0057] The results in Figure 4 showed that compared with the control group, H2O2 caused a decrease in cell viability (46.3 ± 0.3%), with a 9.42 × 10 6 , 1.83×10 7 , 3.71 × 10 7 , 7.36 x 10 7 , 1.47×10 8 , 2.95×10 8After adding Kawamidori exosomes obtained in Preparation Example 1 at 1000particles / mL, the viability returned to 47.9±2.3%, 52.8±1.1%, 59.7±5.1%, 73.9±1.0%, 83.8±1.5%, and 92.8±0.5%, respectively, demonstrating that Kawamidori exosomes have the activity of protecting cells under the platform of oxidative stress-induced HaCaT injury in a dose-dependent manner.

[0058] Images of Kawamidori exosomes protecting skin keratinocytes were also simultaneously observed. Microscopic observation of cell morphology showed that the induction of oxidative stress H2O2 significantly reduced the number of HaCaT cells compared to the control group. However, testing the cell viability of Preparation Example 1 showed that the number of cells recovered as the concentration of Kawamidori exosomes obtained in Preparation Example 1 increased, as shown in Figure 5.

[0059] Example 5: Anti-inflammatory activity of immune cells

[0060] In this example, bacterial lipopolysaccharide (LPS) was used to induce the production of nitric oxide (NO) free radicals in mouse macrophages RAW264.7 to simulate an inflammatory response, and Kawamidori exosomes obtained in Preparation Example 1 were added and then the anti-inflammatory activity of the cells was evaluated. RAW264.7 immune cells were cultured in a DMEM culture medium containing 100 units / mL penicillin, 100 μg / mL streptomycin, and 10% fetal bovine serum in an environment of 5% CO2 and 37°C. First, 2 × 10 4 The cells were cultured in a 96-well culture plate for 24 hours. The next day, the supernatant was removed, and 9.42 × 10 6 , 1.83×10 7 , 3.71 × 10 7 , 7.36 x 10 7 , 1.47×10 8 , 2.95×10 8Cell culture medium containing 0.1g / mL of particles was added and incubated for 1 hour. Subsequently, lipopolysaccharide (LPS) was added to a final concentration of 100nM and incubated in an incubator for 16 hours. On the third day, the supernatant was collected and the NO content was detected by Griess reagent. The cells were incubated with MTT (0.5mg / mL) for 30 minutes and then dissolved in DMSO. The cell viability of each group was measured by OD. 570 The absorbance values ​​were expressed as a percentage after comparing with the control cell group. The results in Figure 6 indicate that co-treatment with LPS at 0.31%, 0.63%, 1.25%, 2.5%, and 5% concentrations of Kawamidori exosomes obtained in Preparation Example 1 did not affect cell viability. In the inflammatory mode (shown in Figure 7), LPS was able to induce NO production (18.3 ± 0.6 μM) compared to the control group (10.7 ± 1.2 μM), and 9.42 × 10 Kawamidori exosomes obtained in Preparation Example 1 were significantly increased. 6 , 1.83×10 7 , 3.71 × 10 7 , 7.36 x 10 7 , 1.47×10 8 , 2.95×10 8 After adding 1000 particles / mL, the NO concentrations were suppressed to 16.4±1.4 μM, 15.4±0.9 μM, 12.7±0.3 μM, 9.8±0.2 μM, 6.5±0.7 μM, and 3.6±0.5 μM, respectively, demonstrating that the Kawamidori exosomes obtained in Preparation Example 1 have anti-inflammatory activity in the inflammatory mode and in a dose-dependent manner.

[0061] Example 6: Safety evaluation

[0062] In this example, a safety evaluation test was conducted using an in vitro bionic skin platform, and a skin irritation test was conducted on Kawamidori exosomes from Preparation Example 1. The skin irritation test included the following procedures. This test was conducted using a human skin model with a detection kit (EpiSkin™ KIT, WPISKIN / S / 13) approved by the Organization for Economic Cooperation and Development (OECD) TG 439. At the time of the test, Kawamidori exosomes from Preparation Example 1 were dissolved in 1x PBS to a final concentration of 5.89x10 8 After adjusting the concentration to 0.15 microparticles / mL, the solution was applied directly to the surface of the skin model. After 15 minutes of incubation, the solution was washed and the skin model was cultured for 42 hours. Finally, the effect of Preparation 1 or the positive control group (5 wt% SDS) on cell activity was evaluated by cell viability analysis (MTT assay). The irritation assessment standard is based on Type 2 of the globally harmonized system of classification and labeling of chemicals (UN GHS), where a cell viability of 50% or less is considered to be skin irritant.

[0063] For test results, see Figure 8, which shows the results of the skin irritation test for Production Example 1, the control group, and the positive control group of the present disclosure. The experimental data are expressed as mean ± SD, and the number of samples was three for each group. After the skin irritation test, the survival rate of the control group was 100.0 ± 1.9%, that of Production Example 1 was 133.2 ± 1.8%, and that of the positive control group was 10.5 ± 0.2%. As can be seen from Figure 8, the Kawamidori exosomes of Production Example 1 were non-irritating in the skin irritation test.

[0064] Furthermore, in this example, a safety evaluation test for eye irritation was also conducted, and an eye irritation test was conducted on Kawamidori exosomes of Production Example 1. This test was based on the OECD TG 492 standard and was conducted using a human corneal epithelial tissue model with a detection kit (LabCyte (trademark) CORNEA-MODEL, Cat. No. 411324). During the test, Kawamidori exosomes of Production Example 1 were dissolved in 1x PBS to a final concentration of 5.89x10 8 After adjusting the concentration to 0.01g / mL, the solution was applied directly to the surface of the corneal epithelial tissue model and allowed to react for 60 minutes, after which it was washed off. The corneal epithelial tissue model was cultured for 24 hours, and finally the effect on cell activity of Preparation 1 or the positive control group (ethanol) was evaluated by cell viability analysis (WST-8 assay kit). The evaluation standard for irritation is based on UN GHS Type 2, where a cell viability of ≦40% is considered to be an eye irritation test.

[0065] For the test results, see Figure 9, which shows the results of the corneal epithelial tissue irritation test for Production Example 1 of the present disclosure, the control group, and the positive control group. The experimental data are expressed as mean ± standard deviation (mean ± SD), and the number of samples was three for each group. After the eye irritation test, as shown in Figure 9, the survival rate of the control group was 100.0 ± 1.8%, that of Production Example 1 was 113.3 ± 13.8%, and that of the positive control group was 18.2 ± 5.1%. As can be seen from Figure 9, the Kawamidori exosomes of Production Example 1 demonstrated no irritation in the eye irritation test.

[0066] Although the present disclosure has been disclosed in the above embodiments, this is not intended to limit the present disclosure, and anyone skilled in the art can make various changes and modifications without departing from the spirit and scope of the present disclosure. Therefore, the scope of protection of the present disclosure should be based on what is defined by the claims attached below.

Claims

1. A composition for strengthening the skin barrier and preventing inflammation, comprising exosomes isolated from Atractylodes mori as an active ingredient.

2. 2. The composition of claim 1, wherein the exosomes are isolated from the aerial parts of Atractylodes macrocarpa.

3. 2. The composition of claim 1, wherein the exosomes have an average particle size of 20 nanometers to 500 nanometers.

4. The exosomes have a concentration of 1×10 ６ Particles / milliliter ~ 1 x 10 １２ 10. The composition of claim 1 comprising particles per milliliter.

5. The composition of claim 1 , wherein the exosomal transmembrane protein comprises CD9.

6. In the use of exosomes for producing a skin barrier strengthening and anti-inflammatory composition, The exosomes are isolated from a green alga,

7. The use according to claim 6 , wherein the exosomal transmembrane protein comprises CD9.

8. The exosomes The process of removing the above-ground parts of the Kawamidori tree, adding a phosphate buffer solution to the aerial parts of the genus Eucalyptus and then grinding the mixture to obtain a crude extract; filtering the crude extract to obtain a crude filtrate; and separating the crude filtrate by tangential flow filtration to obtain the exosomes.

9. The use according to claim 6, wherein the composition is a cosmetic composition, a pharmaceutical composition or a food composition.

10. The exosomes have a concentration of 1×10 ６ Particles / milliliter ~ 1 x 10 １２ 7. The use of claim 6, in the range of particles per milliliter.

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