Anti-aging and antioxidant cosmetic composition comprising broccoli exosomes as active ingredient and functional cosmetics comprising the same

Abstract

The present invention relates to a functional cosmetic composition containing broccoli exosomes as an active ingredient and a functional cosmetic containing the same. The broccoli exosomes have a bilipid membrane structure and are adsorbed into skin cells. The adsorbed exosomes remove reactive oxygen species from skin cells and stimulate various antioxidant signaling mechanisms to exhibit antioxidant efficacy and promote the synthesis of collagen and hyaluronic acid. Therefore, they can be effectively used to produce functional cosmetics for skin anti-aging, skin whitening, wrinkle improvement, and skin cell regeneration.

Description

Detailed Description of the Invention Technical Field

[0002] This application claims priority based on Korean Patent Application No. 10-2020-0099518, filed on August 7, 2020, and all contents disclosed in the specification and drawings of the corresponding application are incorporated into this application.

[0003] The present disclosure relates to a cosmetic composition for anti-aging, anti-oxidation and anti-inflammation and a functional cosmetic containing the same. Background Art

[0004] There are many causes of skin aging, and in particular, reactive oxygen species produced by ultraviolet rays and respiration are considered to be the most important causes of skin aging. Reactive oxygen species are produced during normal metabolic processes and are produced in excess when disease states or physical stress are high. In addition, when exposed to ultraviolet rays, reactive oxygen species are produced. Reactive oxygen species cause damage to skin cells and tissues. Under normal circumstances, they kill abnormal and aging cells to establish a balance, but when produced in excess, they destroy the skin's antioxidant defense network, disrupt lipid peroxidation, protein oxidation and matrix components, and damage skin proteins such as collagen, elastin, etc., and therefore, they cause skin aging such as loss of elasticity, wrinkles, pigmentation, etc.

[0005] Skin proteins, including those in the connective tissue of the dermis, play an important role in maintaining skin elasticity and moisture, and this can be attributed to the fact that proteins are essential for cell construction and production. Recently, research has demonstrated a positive correlation between protein intake and increased skin moisture, and from a nutritional perspective, it can be inferred that high-quality protein intake is closely related to skin health. However, the effect of protein intake through food on skin is only modest.

[0006] Approximately 500 or more plant extracts with specific pharmacological activities, such as antioxidant and anti-aging properties, are known in Korea. However, the effects of most of these extracts on the skin have not been confirmed, and numerous ingredients are present in varying ratios in plant extracts, making the discovery of active substances with specific effects difficult. Furthermore, among plant components, active substances that exhibit effects may act as a single component alone, but these effects may also be achieved through the interaction of several components. Furthermore, many conventional plant extracts are known to have difficulty crossing the skin barrier, and their effects are limited by simply applying them to the skin.

[0007] Throughout this specification, many documents are referenced and their citations are indicated. The disclosures of the cited documents are incorporated into this specification in their entirety by reference to more clearly describe the level of technology to which the present disclosure belongs and the content of the present disclosure. Summary of the Invention

[0008] Technical issues

[0009] The present disclosure relates to the anti-aging, antioxidant and anti-inflammatory effects of exosomes isolated and purified from broccoli, and one object of the present disclosure is to provide a cosmetic composition using broccoli exosomes having anti-aging, antioxidant and anti-inflammatory functions as an active ingredient.

[0010] Another object of the present disclosure is to provide functional cosmetics for skin protection, wrinkle improvement, skin whitening, etc., prepared by using the composition as a cosmetic raw material.

[0011] Another object of the present disclosure is to provide a method for skin antioxidant comprising applying a cosmetic composition comprising broccoli exosomes to a subject.

[0012] Another object of the present disclosure is to provide cosmetic uses of a cosmetic composition comprising broccoli exosomes for skin antioxidant effects.

[0013] Another object of the present disclosure is to provide cosmetic uses of a cosmetic composition comprising broccoli exosomes for skin whitening, wrinkle improvement, and skin cell regeneration.

[0014] More specifically, the present disclosure aims to provide the following embodiments.

[0015] Embodiment 1. A composition or cosmetic composition comprising broccoli exosomes; a method for skin aging prevention, skin anti-oxidation, skin whitening, skin wrinkle improvement, skin cell regeneration, skin moisturizing, skin barrier improvement or skin anti-inflammation, the method comprising applying a composition comprising broccoli exosomes to a subject in need thereof or comprising applying isolated broccoli exosomes to a subject in need thereof; use or cosmetic use of isolated broccoli exosomes or a composition comprising broccoli exosomes for exhibiting cosmetic efficacy selected from skin aging prevention, skin anti-oxidation, skin whitening, skin wrinkle improvement, skin cell regeneration, skin moisturizing, skin barrier improvement and skin anti-inflammation; or use of a composition comprising broccoli exosomes for preparing a cosmetic product for skin aging prevention, skin anti-oxidation, skin whitening, skin wrinkle improvement, skin cell regeneration, moisturizing, skin barrier improvement or skin anti-inflammation, or use of isolated broccoli exosomes for preparing a cosmetic product for skin aging prevention, skin anti-oxidation, skin whitening, skin wrinkle improvement, skin cell regeneration, moisturizing, skin barrier improvement or skin anti-inflammation.

[0016] Embodiment 2. The composition; method; or use or cosmetic use according to embodiment 1, wherein the broccoli exosomes are derived from at least one member selected from the group consisting of broccoli flowers, broccoli stems, and broccoli sprouts.

[0017] Embodiment 3. The composition according to any one of the preceding embodiments; method; or use or cosmetic use, wherein the broccoli exosomes are present in an amount of 10 6 to 10 13 The amount of particles is included.

[0018] Embodiment 4. The composition; method; or use or cosmetic use according to any one of the preceding embodiments, wherein the composition is a formulation selected from the group consisting of an aqueous solution, a suspension, an emulsion, a cream, a frozen solution, a spray-dried powder, and a freeze-dried powder.

[0019] Embodiment 5. The composition; method; or use or cosmetic use according to any one of the preceding embodiments, wherein the broccoli exosomes comprise polyphenols.

[0020] Embodiment 6. The composition; method; or use or cosmetic use according to any one of the preceding embodiments, wherein the cosmetic composition has anti-aging or antioxidant uses.

[0021] Embodiment 7. A cosmetic comprising the composition according to any one of the preceding embodiments.

[0022] Embodiment 8. The cosmetic according to any one of the preceding embodiments, wherein the cosmetic is a preparation selected from the group consisting of a spray, a serum, a nourishing lotion, a soft lotion, a soft water, an emulsion, a skin lotion, a skin softener, a firming agent, an astringent, a lotion, a milky lotion, a moisturizing lotion, a nourishing lotion, a massage cream, a nourishing cream, a moisturizing cream, a hand cream, a foundation, a powder, a makeup base, a serum, a nourishing serum, a peel-off pack, a soap, a cleansing foam, a cleansing lotion, a cleansing cream, a body wash, a body cleanser, a facial wash, a treatment, a cosmetic liquid, a cosmetic mask, an ointment, a gel, a liniment, a liquid, a patch, a spray, a bath preparation, a sunscreen, a suntan oil, and a hair care product.

[0023] Embodiment 9. The cosmetic according to any one of the preceding embodiments, wherein the cosmetic is a functional cosmetic product having at least one effect selected from the group consisting of skin aging prevention, skin whitening, wrinkle improvement, skin cell regeneration, moisturizing, skin barrier improvement, and anti-inflammation.

[0024] Embodiment 10. A method of preparing the composition or cosmetic composition according to any one of the preceding embodiments, comprising crushing broccoli; performing centrifugation; and performing tangential-flow filtration (TFF).

[0025] Other objects and advantages of the present disclosure will become more apparent from the following detailed description of the invention, the claims and the accompanying drawings.

[0026] Technical Solution

[0027] One aspect of the present disclosure is to provide a cosmetic composition comprising broccoli sprout exosomes as an active ingredient.

[0028] The term "broccoli exosomes" as used herein refers collectively to exosomes or extracellular endoplasmic reticulum, comprising exosomes extracted from broccoli having a size of 50 nm to 300 nm, preferably a size of 50 nm to 200 nm. The broccoli exosomes form particles of uniform size of 50 nm to 200 nm in the form of a lipid bilayer.

[0029] Various conventional plant extracts are known to contain anti-aging and antioxidant substances, but they have the following problems: they have difficulty passing the skin barrier in the form of water-soluble extracts, and the active ingredients are easily destroyed by exposure to the extraction solvent during the extraction process, making them difficult to use as cosmetic raw materials. On the other hand, exosomes are nanoparticles composed of a lipid bilayer and are absorbed into skin cells through receptor-mediated endocytosis and membrane fusion. In addition, they are very advantageous for commercial use in the cosmetics industry because the active ingredients contained are not directly exposed to the outside, so they are not easily degraded and can maintain biological activity for a long time.

[0030] In the present disclosure, broccoli exosomes, through the anti-aging and antioxidant components derived from broccoli (which is a raw material containing broccoli sprouts), have the function of inhibiting reactive oxygen species that cause skin aging, pigmentation, and inflammation, and thus have anti-aging and / or antioxidant uses. Specifically, broccoli exosomes, which contain polyphenols and antioxidant enzyme promoters, inhibit accumulated reactive oxygen species, thereby demonstrating effects such as skin regeneration, wrinkle improvement, whitening, and inflammation relief.

[0031] The raw material plant broccoli of the present invention includes cabbage (Brassica Oleracea, Brassica Oleracea L.), which is a species of Brassica. As the raw material plant part, the whole broccoli or at least a part including flowers, stems, buds, etc. can be mixed and used.

[0032] In the present disclosure, the broccoli / broccoli sprout exosomes extracted from broccoli / broccoli sprouts can be prepared at a concentration of 1×10 6 to 1×10 13 , preferably 1×10 7 to 1×10 13 The content of each particle is included.

[0033] Broccoli / broccoli sprout exosomes themselves have anti-aging and antioxidant functions similar to superoxide dismutase (SOD), and this promotes the decomposition of reactive oxygen species existing outside cells and accumulated within cells, and exhibits the effects of promoting collagen production, promoting hyaluronic acid production, inhibiting melanin synthesis, and inhibiting the production of inflammatory substances.

[0034] Furthermore, the supply of high-quality protein is closely related to increased skin moisture and skin health, and the broccoli exosomes of the present disclosure can effectively deliver plant protein to the skin by containing protein at a high concentration of 10 μg / mL to 1,000 μg / mL, preferably 100 μg / mL to 1,000 μg / mL.

[0035] In one embodiment, the cosmetic composition for antioxidant comprising broccoli / broccoli sprout exosomes as an active ingredient may be provided in a formulation selected from the group consisting of aqueous solution, suspension, emulsion, cream, frozen solution, spray-dried powder, and freeze-dried powder.

[0036] Another aspect of the present disclosure is to provide a cosmetic comprising a cosmetic composition for anti-oxidation containing broccoli / broccoli sprout exosomes as an active ingredient.

[0037] The cosmetics can be applied as needed and mixed with general components for preparing general skin cosmetics, such as oil, water, surfactants, moisturizing agents, lower alcohols, thickeners, chelating agents, pigments, preservatives, flavoring agents, etc. In addition, the cosmetics can be prepared in a preparation selected from the following: sprays, serums, nourishing lotions, soft lotions, soft water, emulsions, skin lotions, skin softeners, skin tightening agents, astringents, lotions, emulsion lotions, moisturizing lotions, nourishing lotions, massage creams, nourishing creams, moisturizing creams, hand creams, foundations, powders, cosmetic bases, essences, nourishing essences, peel-off masks, soaps, cleansing foams, cleansing lotions, cleansing creams, body lotions, body cleansers, facial lotions, care agents, cosmetic liquids, cosmetic masks, ointments, gels, liniments, liquids, patches, sprays, bath preparations, sunscreens, sunscreen oils, and hair care products, and they can contain various conventional carriers and additives that are suitable for their various preparations and are well known in the art.

[0038] Such cosmetics of the present disclosure may be functional cosmetics having at least one effect selected from the group consisting of skin aging prevention, skin whitening, wrinkle improvement, and skin cell regeneration.

[0039] Another aspect of the present disclosure is to provide a method for preparing a cosmetic composition for anti-aging and anti-oxidation comprising broccoli / broccoli sprout exosomes as an active ingredient, the method comprising crushing broccoli; performing centrifugation; and performing tangential flow filtration (TFF).

[0040] The process of crushing the raw material plant may include a process of mechanically crushing a mixture in which the raw material and the buffer solution are mixed in a weight ratio ranging from 1:1 to 1:10, and this may be performed by mechanically crushing the plant mixture via a rotating blade at a speed of 10 rpm to 1,000 rpm.

[0041] The centrifugation method can be selected from low-speed centrifugation, high-speed centrifugation, ultracentrifugation and combinations thereof, and preferably, it can include ultracentrifugation. Low-speed centrifugation can achieve a speed of 6,000 rpm (6,000 x g) or less, and is mainly used for samples such as cells or cell nuclei that are easily precipitated by centrifugation, and high-speed centrifugation has a maximum speed of about 20,000 rpm to 25,000 rpm (60,000 x g), and ultracentrifugation means a centrifugation method with a maximum speed of about 40,000 rpm to 80,000 rpm (600,000 x g).

[0042] Tangential flow filtration (TFF) is a filtration method in which a solution flows in a direction perpendicular to a filter membrane, thereby filtering out small-sized impurities present in the solution and separating large-sized exosomes. Compared with conventional filtration methods, it can minimize the adsorption of exosomes to the pores of the filter filter or the clogging of the membrane pores, and therefore, it is easy to apply process amplification and cGMP (cosmetic good manufacturing procedure, cosmetics manufacturing procedure) processes. Tangential flow filtration can be at least one selected from hollow fiber TFF and membrane TFF capable of ultrafiltration, and preferably, it can use a TFF filter with a molecular weight cutoff (molecular weight cutoff, MWCO) of 100,000Da to 500,000Da.

[0043] Therefore, broccoli / broccoli sprout exosomes according to the present disclosure can be isolated from broccoli and broccoli sprouts by mechanical crushing, stepwise ultracentrifugation and tangential flow filtration (TFF) methods, and these exosomes contain a large amount of polyphenols with anti-aging and antioxidant functions.

[0044] Beneficial effects

[0045] Broccoli exosomes have a lipid bilayer structure and are adsorbed into skin cells. The adsorbed polyphenolic components of broccoli exosomes remove reactive oxygen species within skin cells and stimulate antioxidant signaling mechanisms. Therefore, they have a variety of functional cosmetic uses, such as skin whitening, wrinkle improvement, skin aging prevention, skin barrier improvement, inflammation relief, and skin cell activation. Furthermore, the excellent stability of broccoli exosomes allows for their use in various types of functional cosmetics, such as aqueous solutions, frozen solutions, and freeze-dried powders. BRIEF DESCRIPTION OF THE DRAWINGS

[0046] Figure 1 is a schematic diagram of the process of the method for isolating plant exosomes.

[0047] Figure 2 These are the results of morphological analysis (TEM) of broccoli sprout exosomes.

[0048] Figure 3 These are the results of characterization of broccoli sprout exosomes (NTA and protein assay).

[0049] Figure 4 These are the results of an analysis of the polyphenol content and copper reducing capacity of broccoli sprout exosomes.

[0050] Figure 5 This is the result of evaluating the melanin formation inhibitory ability of broccoli sprout exosomes.

[0051] Figure 6 These are the results of evaluation of the melanin formation inhibitory ability of broccoli sprout exosomes in a cell model in which melanin formation is promoted by α-MSH.

[0052] Figure 7 This is the result of evaluating the collagen-producing capacity of broccoli sprout exosomes.

[0053] Figure 8 This is the result of evaluating the hyaluronic acid-producing capacity of broccoli sprout exosomes.

[0054] Figure 9 This is the evaluation result of the NO production inhibitory ability of broccoli sprout exosomes.

[0055] Figure 10 The results are the evaluation results of the cytotoxicity of broccoli sprout exosomes on human fibroblasts (HDF), B16F10 melanoma cells and mouse macrophages (RAW 264.7). DETAILED DESCRIPTION

[0056] Hereinafter, the present disclosure will be described in more detail by way of examples. These examples are only intended to illustrate the present disclosure in more detail, and it will be apparent to those skilled in the art to which the present disclosure pertains that the scope of the present disclosure is not limited by these examples.

[0057] Example

[0058] <Example 1> Plant exosome separation

[0059] Broccoli and broccoli sprouts, aloe vera peel, and spinach were used as raw materials, and plant exosomes were isolated from them using ultracentrifugation and tangential flow filtration (TFF). Specifically, the cleaned plant material was crushed and homogenized with phosphate-buffered saline (PBS) using a blender at a weight ratio of 1:1 (weight / weight) to 1:10 (weight / weight). The supernatant was collected by centrifugation at 1,000 × g for 10 minutes and sequentially centrifuged at 2,000 × g for 20 minutes, 3,000 × g for 30 minutes, and 10,000 × g for 60 minutes. The supernatant was then ultracentrifuged at 100,000 × g for 70 minutes at 4°C, and the vacuoles were removed through this process. Finally, the supernatant was discarded, and the remaining pellet was suspended and purified using a TFF system. Specifically, the exosomes were purified in a TFF system by using a filter with a molecular weight cutoff of 100 kDa to 500 kDa, and other impurity particles smaller than the filter pores were removed, and the solution containing the exosomes was concentrated. The isolated exosomes were stored frozen at -70°C or lower until use ( Figure 1 ).

[0060] <Example 2> Evaluation of exosome protein content

[0061] Micro BCA from Thermo Scientific TMThe protein assay kit measures the protein content contained in broccoli sprout exosomes, aloe vera exosomes, and spinach exosomes. Two groups of exosomes were used for comparative studies, one group consisting of exosome samples immediately after isolation and the other consisting of exosome samples stored at -70°C for at least one month. Specifically, using a pipette, 1.0 mL of each standard and sample was taken out of the test tube, and 1.0 mL of WR was added to each tube and then mixed. The tubes were capped and incubated in a water bath at 37°C for 30 minutes, then cooled at room temperature. The instrument was zeroed in a cuvette filled with distilled water, the spectrophotometer was set to 562 nm, and the absorbance of the sample was measured within 10 minutes. A standard curve was prepared by measuring the total protein concentration relative to the protein standard and plotting the average blank-corrected 562 nm readings for each standard. Using the standard curve, the protein content of the samples was measured, and the results are shown in Table 1 below.

[0062] [Table 1]

[0063]

[0064] As a result of the experiment, it was confirmed that the broccoli sprout exosomes contained 100 μg / mL or more of protein per 1 mL unit volume even after thawing. Additional property evaluations were performed using the broccoli sprout exosomes confirmed to have a high protein content.

[0065] <Example 3> Evaluation of the characteristics of broccoli sprout exosomes

[0066] For the broccoli sprout exosomes isolated in Example 1, additional exosome property evaluation was performed by the following method. As for the shape of the exosomes, spherical microstructures ( ) of 200 nm or less were confirmed by scanning electron microscopy (TEM) (scale bar 50 nm). Figure 2 The microparticle size was analyzed by nanoparticle tracking analysis (NTA) and determined to be between 50 nm and 300 nm ( Figure 3 ). By nanoparticle tracking analysis (NTA), it was determined that 1×10 8 to 1×10 13 The exosome concentration ( Figure 3 ). Micro BCA from Thermo Scientific was used. TM The protein content in the exosomes was further measured several times using a protein assay kit and determined to contain 10 μg / mL to 1,000 μg / mL of protein per 1 mL unit volume ( Figure 3 ).

[0067] <Example 4> Evaluation of polyphenol content of broccoli sprout exosomes

[0068] The polyphenol content of the broccoli sprout exosomes isolated in Example 1 was evaluated by the following method. The polyphenol content of the broccoli sprout exosomes was measured using a phenolic compound assay kit from BioVision, and the measurement method was performed according to the kit manufacturer's protocol. 8 Each broccoli efflux contains 50 pmol to 500 pmol of polyphenols ( Figure 4 ).

[0069] <Example 5> Evaluation of the Antioxidant Activity of Broccoli Sprout Exosomes

[0070] The polyphenol content of the broccoli sprout exosomes isolated in Example 1 was evaluated by the following method. TM The total antioxidant capacity (TAC) assay kit was used to evaluate the antioxidant capacity of broccoli exosomes, and the measurement method was performed according to the kit manufacturer's protocol. As a result of the evaluation, it was determined that broccoli sprout exosomes had the ability to reduce 10 μM to 100 μM of Cu(II) ions to Cu(I) ions per 1 mL unit volume ( Figure 4 ).

[0071] <Example 6> Evaluation of the in vitro whitening efficacy of broccoli sprout exosomes

[0072] In order to evaluate the antioxidant efficacy of the broccoli sprout exosomes isolated in Example 1, the whitening efficacy of the exosomes was evaluated. 3 to 1×10 4 Skin melanoma (B16F10) cells were treated and, after 3 days of culture, intracellular and extracellular melanin content was measured. To determine the intracellular melanin content, the absorbance of the solution at 475 nm was measured using a UV-visible spectrophotometer after destruction and lysis of the cells. For extracellular melanin, after collecting the cell culture solution for 3 days of culture, the absorbance of the solution at 490 nm was measured using a UV-visible spectrophotometer. As a negative control group, growth medium (GM) without exosomes was used, and as a positive control group, 0.5 mM arbutin, which is widely used in whitening efficacy evaluation, was used.

[0073] Determined at 5×10 7 or greater, broccoli sprout exosomes inhibited extracellular and intracellular melanin formation, and in particular, when 8 When treated under the condition of Figure 5).

[0074] The whitening efficacy of the broccoli sprout exosomes isolated in Example 1 was further evaluated using a model in which skin melanoma (B16F10) cells were treated with 100 nM α-MSH to promote melanin synthesis and secretion. After treating the cell culture solution containing α-MSH and broccoli exosomes, they were cultured for 3 days, and the total content of intracellular and extracellular melanin was measured using a UV-visible spectrophotometer. For protein content measurement, after quantification according to the Bradford method, the amount of melanin was converted into the amount of melanin per protein. As a negative control group, a cell culture medium without exosome treatment was used, and as a positive control group, 0.36 mM arbutin, which is widely used for whitening efficacy evaluation, was used. At 2 × 10 7 or greater, broccoli sprout exosomes inhibited total melanin formation (p<0.05) ( Figure 6 ).

[0075] <Example 7> Evaluation of in vitro collagen formation of broccoli sprout exosomes

[0076] In order to evaluate the efficacy of the broccoli sprout exosomes isolated in Example 1 on collagen formation, the exosomes were treated to human dermal fibroblasts (HDF) to determine the amount of intracellular collagen produced. A cell culture solution containing broccoli sprout exosomes was treated to HDF cells, and after culturing them for 2 days, the amount of procollagen was measured using the supernatant by centrifugation. For the amount of procollagen, the absorbance at 450 nm was measured using a procollagen type I C-peptide (PIP) ELISA kit. As a negative control group, a cell culture solution (GM) without any treatment was used, and as a positive control group, TGF-β1, which is known to induce collagen production, was used. It was determined that compared with the negative control group, at 1.3×10 6 to 1×10 7 At the concentration of 1.57kJ / cm2, broccoli sprout exosomes had a significant effect on increasing the production of intracellular collagen (p<0.05) ( Figure 7 ). This had an efficacy similar to that of the positive control, TGF-β1.

[0077] <Example 8> Evaluation of hyaluronic acid production in vitro by broccoli sprout exosomes

[0078] In order to evaluate the efficacy of the broccoli sprout exosomes isolated in Example 1 on hyaluronic acid production, the exosomes were treated to human dermal fibroblasts (HDF) to determine the amount of intracellular hyaluronic acid produced. A sample containing broccoli sprout exosomes was treated to HDF, and after culturing it for 2 days, the supernatant was collected by centrifugation and used as a test sample to measure the amount of hyaluronic acid produced. In order to determine the amount of hyaluronic acid produced, the absorbance at 450 nm was measured using a human hyaluronic acid (HA) ELISA kit. As a negative control group, growth medium (GM) without exosomes was used, and as a positive control group, retinoic acid for promoting hyaluronic acid production was used. It was determined that compared to the negative control group, at 2.5×10 6 to 1×10 7 At the concentration of 1.57kcal, broccoli sprout exosomes had a significant effect on increasing the production of intracellular hyaluronic acid (p<0.05) ( Figure 8 ). This had an efficacy similar to that of the positive control, retinoic acid.

[0079] <Example 9> Evaluation of Nitric Oxide Production in Vitro by Broccoli Sprout Exosomes

[0080] In order to evaluate the efficacy of the broccoli sprout exosomes isolated in Example 1 in inhibiting nitric oxide (NO) production, the exosomes were treated with mouse macrophages (RAW 264.7 cells) to determine whether they inhibited NO production. 5 After culturing RAW264.7 cells, the culture medium was replaced with a new cell culture solution containing 1 μg / mL lipopolysaccharide (LPS) and broccoli sprout exosomes and then incubated for 24 hours. At 24 hours, the culture solution was reacted with a NO detection reagent for 10 minutes, and the absorbance of the solution at 540 nm was measured. The amount of NO produced was assessed by correcting for total protein. As a positive control, 7.8 μg / mL of dexamethasone, which has an anti-inflammatory effect, was used.

[0081] Determine the broccoli sprout exosomes at 2×10 7 or more had the effect of inhibiting NO production (p<0.05) ( Figure 9 ). Preferably, at 2×10 7 to 4×10 7 In the presence of a single exosome, it effectively inhibited cellular NO production with a potency similar to that of the positive control, dexamethasone.

[0082] <Example 10> Evaluation of cytotoxicity of broccoli sprout exosomes

[0083] To determine the cytotoxicity of the broccoli sprout exosomes isolated in Example 1, the cell viability of two human cells and mouse macrophages was assessed using the MTT assay after treatment of the exosomes.

[0084] In 5×10 4 After inoculating human dermal fibroblasts at a concentration of 1.5×10 cells / well and culturing them for 24 hours, broccoli sprout exosomes were treated and then further cultured for 24 hours. 4 After inoculating B16F10 melanoma cells at a concentration of 1.8×10 cells / well and culturing them for 24 hours, broccoli exosomes were treated and then cultured for 72 hours. 5 After inoculating mouse macrophages (RAW 264.7) at a concentration of 10 cells / well and culturing them for 20 hours, broccoli exosomes were treated and then cultured for 24 hours. MMT reagent was added to each well at a concentration of 0.05% of the cell culture medium and then incubated for 4 hours. After removing the culture solution, DMSO was inoculated at 1 mL / well and stirred for 10 minutes. The DMSO solution was then recovered and the absorbance at 540 nm was measured. Cell viability was expressed as a percentage compared to the exosome-untreated group ( Figure 10 As a result of the experiment, it was determined that broccoli sprout exosomes showed no cytotoxicity at all concentrations.

Claims

1. Use of isolated broccoli exosomes for the preparation of products for skin whitening or hyaluronic acid production, wherein the broccoli exosomes are isolated as active ingredients from a broccoli raw material comprising broccoli sprouts, and The product is a cosmetic product, wherein the broccoli exosomes are present in an amount of 10 6 to 10 13 An amount of 100 particles is contained in the composition, and the cosmetic composition is contained in the product.

2. The use according to claim 1, wherein the composition is formulated into one selected from the group consisting of an aqueous solution, a suspension, an emulsion, a cream, a frozen solution, a spray-dried powder, and a freeze-dried powder.

3. The use according to claim 1, wherein the broccoli exosomes comprise polyphenols.

4. The use according to claim 1, wherein the cosmetic product is a preparation selected from the group consisting of serums, nourishing lotions, softening lotions, emulsions, skin softeners, skin tighteners, astringents, lotions, massage creams, nourishing creams, moisturizing creams, hand creams, foundations, powders, makeup bases, essences, peel-off masks, soaps, cleansing foams, cleansing creams, care agents, makeup lotions, makeup masks, ointments, gels, liniments, patches, sprays, bath preparations, sunscreens, suntan oils, and hair care products.

5. The use according to claim 1, wherein the cosmetic product is a formulation selected from the group consisting of a cleansing lotion and a facial lotion.

6. The use according to claim 1, wherein the cosmetic product is a formulation selected from the group consisting of a spray and a body cleanser.

7. A method for preparing a product for skin whitening or hyaluronic acid production, comprising: Crush the broccoli; Perform centrifugation; Tangential flow filtration (TFF) was performed to separate broccoli exosomes; as well as causing isolated broccoli exosomes to be included in the product, wherein the broccoli exosomes are isolated as active ingredients from a broccoli raw material comprising broccoli sprouts, and The product is a cosmetic product.

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