WO2019088545A1 - Kit d'exosomes et procédé d'amélioration de la perméation transdermique d'exosomes l'utilisant - Google Patents

Kit d'exosomes et procédé d'amélioration de la perméation transdermique d'exosomes l'utilisant Download PDF

Info

Publication number
WO2019088545A1
WO2019088545A1 PCT/KR2018/012549 KR2018012549W WO2019088545A1 WO 2019088545 A1 WO2019088545 A1 WO 2019088545A1 KR 2018012549 W KR2018012549 W KR 2018012549W WO 2019088545 A1 WO2019088545 A1 WO 2019088545A1
Authority
WO
WIPO (PCT)
Prior art keywords
skin
exosome
exosomes
battery
patch
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/KR2018/012549
Other languages
English (en)
Korean (ko)
Inventor
이용원
조병성
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Biosensor Laboratories Inc
Exocobio Inc
Original Assignee
Biosensor Laboratories Inc
Exocobio Inc
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Biosensor Laboratories Inc, Exocobio Inc filed Critical Biosensor Laboratories Inc
Publication of WO2019088545A1 publication Critical patent/WO2019088545A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Images

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
    • A61K8/98—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution of animal origin
    • A61K8/981—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution of animal origin of mammals or bird
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
    • A61K35/28—Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/02—Cosmetics or similar toiletry preparations characterised by special physical form
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/02—Cosmetics or similar toiletry preparations characterised by special physical form
    • A61K8/0212—Face masks
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/02—Cosmetics or similar toiletry preparations characterised by special physical form
    • A61K8/14—Liposomes; Vesicles
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
    • A61K8/98—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution of animal origin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/10—Dispersions; Emulsions
    • A61K9/127—Synthetic bilayered vehicles, e.g. liposomes or liposomes with cholesterol as the only non-phosphatidyl surfactant
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00—Preparations for care of the skin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
    • A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
    • A61K2800/87—Application Devices; Containers; Packaging

Definitions

  • the present invention relates to an exosomal kit for the percutaneous permeation enhancement of exosomes comprising a mask pack, a mask sheet or a patch applied or immersed with a composition comprising an exosome as an active ingredient, and an iontophoresis device will be.
  • the present invention also relates to a method for enhancing transdermal permeability of exosome and its application, which enables the exosome to pass through the skin barrier effectively and to be deeply transferred into the skin using the exosome kit.
  • the skin has a three-layer structure of epidermis, dermis, and subcutaneous fat.
  • the stratum corneum of the epidermis which is located at the outermost part of the skin, suppresses moisture evaporation of the skin and acts as a skin barrier.
  • the skin barrier of the stratum corneum has a problem of impairing the penetration of the active ingredient when the functional cosmetic or the medicinal product is applied to the skin, thereby deteriorating the efficacy of the functional cosmetic or the medicinal product.
  • a transdermal delivery system is an effective ingredient applied to the surface of the skin to easily pass through the skin barrier and deliver it deeply into the skin.
  • transdermal delivery systems include liposomes. Liposomes are composed of lipid bilayers that are structurally similar to cell membranes or intercellular lipids of the stratum corneum, so they can be fused with cell membranes, effectively transferring the active ingredients inside the liposomes into the skin. Artificially synthesized liposomes, however, have limited stability and limited skin penetration.
  • Ethosomes are made by dissolving phospholipids in ethanol known as skin permeation enhancers. Ethanol ethanol weakens the intercellular lipid membranes of the skin's horny layer, increasing the skin's absorption rate of the active ingredient and making the membrane of the endoplasmic reticulum itself flexible. This feature allows the etosome to deliver effective ingredients to the deeper and deeper part of the active ingredient in the skin.
  • ethosomes also have problems such as particle aggregation due to curing of nanoparticles and layer separation phenomenon.
  • exosome which has intercellular signal transduction
  • Extracellular vesicles Cells release various membrane-type vesicles in the extracellular environment, and these release vesicles are commonly referred to as extracellular vesicles (EVs).
  • the extracellular endoplasmic reticulum is sometimes referred to as cell membrane-derived endoplasmic reticulum, ectosomes, shedding vesicles, microparticles, exosomes and, in some cases, differentiated from exosomes.
  • Exosome is an endoplasmic reticulum of several tens to several hundreds of nanometers in size, composed of double lipid membranes identical to the cell membrane structure, and contains proteins, nucleic acids (mRNA, miRNA, etc.) called exosomal cargo inside.
  • Exosomal cargo contains a wide range of signaling factors, which are known to be specific for cell types and differentially regulated by the environment of the secretory cells.
  • Exosome is an intercellular signaling mediator that is secreted by the cell, and various cell signals transmitted through it regulate cell behavior including activation, growth, migration, differentiation, de-differentiation, apoptosis, and necrosis of target cells It is known.
  • Exosomes contain specific genetic material and bioactivity factors depending on the nature and condition of the derived cells.
  • the proliferating stem cell-derived exosomes regulate cell behavior such as cell migration, proliferation and differentiation, and reflect the characteristics of stem cells involved in tissue regeneration (Nature Review Immunology 2002 (2) 569-579).
  • the inventors of the present invention have developed an exosomic kit for transcutaneous permeation enhancement of exosomes which greatly promotes transdermal permeation of exosomes while repeatedly studying the new application fields of exosomes and medical or cosmetic techniques Thus completing the present invention.
  • It is an object of the present invention to provide an exosome kit for enhancing transcutaneous permeation of exosomes comprising a mask pack, a mask sheet or a patch applied or immersed with a composition containing an exosome as an active ingredient, and an iontophoresis device, .
  • the present invention provides a method for treating a transplant of an exosomes comprising an iontophoresis device and a mask pack, a mask sheet or a patch, on which a composition containing an exosome as an active ingredient is applied or immersed, Thereby providing an exosome kit for enhancing permeability.
  • exosomes refers to an endoplasmic reticulum of several tens to several hundred nanometers (preferably about 30 to 200 nm) in size, consisting of double lipid membranes identical in structure to the cell membrane The particle size of the exosome can be varied according to the cell type, the separation method and the measurement method) (Vasiliy S. Chernyshev et al., "Size and shape characterization of hydrated and desiccated exosomes", Anal Bioanal Chem, DOI 10.1007 / s00216-015-8535-3). Exosomes contain proteins called exosomal cargo (cargo), nucleic acids (mRNA, miRNA, etc.).
  • Exosomal cargo contains a wide range of signaling factors, which are known to be specific for cell types and differentially regulated by the environment of the secretory cells.
  • Exosome is an intercellular signaling mediator that is secreted by the cell, and various cell signals transmitted through it regulate cell behavior including activation, growth, migration, differentiation, de-differentiation, apoptosis, and necrosis of target cells It is known.
  • biological solution means a liquid solution of biogenic origin, in which the exosome is dispersed, suspended, precipitated, suspended or mixed, and includes, for example, a cell culture solution, a cell culture supernatant, Cell culture supernatant, whole blood, serum, cord blood, plasma, multiple fluid, brain and cerebrospinal fluid, placenta extract, and bone marrow aspirate.
  • a cell culture solution a cell culture supernatant, Cell culture supernatant, whole blood, serum, cord blood, plasma, multiple fluid, brain and cerebrospinal fluid, placenta extract, and bone marrow aspirate.
  • a “ biological solution” may be cultured or incubated under conditions that release and / or secrete exosomes, and may be frozen and thawed.
  • exosome is intended to mean an exosome that is secreted from cells of various animals, plants, bacteria, fungi, algae and the like, preferably stem cells, (E. G., Exosome-like vesicles) having a nano-sized bezacl structure and a composition similar to exosome.
  • exosome used in the present invention can be applied to skin and various exosomes which are used in the art or can be used in the future can be used as long as they do not cause adverse effects on the human body. Therefore, it should be understood that the exosome isolated according to the separation method of the following embodiments should be understood as an example of exosome that can be used in the present invention, and the present invention is not limited thereto.
  • the term " improvement of skin condition" is a positive change visually and / or tactually perceptible to the appearance and / or feel of the skin, including skin tissue regeneration, Improving the skin condition, preventing aging of the skin, or restoring the skin condition to a mild condition, an improvement condition or a normal condition.
  • the term " improvement of skin condition" refers to alleviation, improvement or elimination of skin defects, skin wrinkles, reduced skin elasticity, wrinkles, wrinkles, rough and deep wrinkles, cracks, bumps, ; Skin thickening (e.g., removal of the epidermis, dermis, subcutaneous layer, nail and stratum corneum of the skin); Prevention of skin or hair elasticity loss due to loss, damage and / or inactivation of functional skin elastin; Reduction of cellulite; Discoloration of pale skin, spots, freckles, dullness, skin, hair or nails, such as blackness, dark circles under eyes, erythema, dots, café aureus, becker spots, osteoblast, pigmentary nevus, epidermal nevus, melanin pigment spot Alleviating, improving or eliminating pigmentation, capillary vasodilation or discoloration caused by spider blood vessels; Alleviating or preventing skin dryness and brittleness.
  • Skin thickening e.g., removal of the epider
  • the term " skin aesthetic" is intended to reduce, alleviate, or ameliorate skin aging such as skin sagging, dented cheeks, depressed eyes, wrinkles, reduced skin elasticity, Restoring the skin condition to a normal state; Skin, skin regeneration, skin whitening, freckles, dark circles, spots, tattoos, acne, elastin layer marks, stretch marks, scarring, enlarged pores, pale, spots, black circles, dark circles, erythema, Removal or correction of various skin defects such as spots, osteoblasts, pigmented nevi, epidermal nevus, melanin pigment spots, and pigmentation; Hair loss enhancement, scalp management, cellulite reduction, facial reduction, contour correction, soft tissue defect correction, tissue enlargement, volume-up, skin swelling, skin moisturizing and soothing, redness and blood vessels Reduction or improvement of diabetes mellitus.
  • skin beauty is not limited to the above, and includes, for example, removing or correcting various skin imperfections.
  • the term "treatment of skin diseases" refers to the treatment of facial flushing, spots, freckles, dullness, dark circles, dark circles, erythema, dots, café au lait, becker dots, Or ameliorating or alleviating pigmented skin lesions such as pigmentation, or restoring skin conditions that are deteriorated by such pigmented skin lesions to a normal state; Relieving, alleviating or ameliorating vascular skin lesions such as flame nevus, capillary vasodilatation, injection, hemangiomas, vesicular blebs, etc., or to restore normalized skin conditions deteriorated by such vascular skin lesions; Such as contact dermatitis, irritant contact dermatitis, allergic contact dermatitis, phototoxic and photoallergic contact dermatitis, contact urticaria, atopic dermatitis, seborrheic dermatitis, self-sensitizing dermatitis, autoimmune progesterone dermatitis, dermatiti
  • iontophoresis refers to a method in which an ionized active ingredient is allowed to permeate through the skin by an electrical repulsive force by applying a minute current to the skin to which the effective substance is applied, .
  • the iontophoresis used in one embodiment of the present invention is a method in which a current flows from an external power source to an electrode patch on the skin to introduce minute current into the skin, A method in which minute current is introduced, a method in which minute current is introduced into skin through a patch equipped with a reversed electrodialysis means for generating current through a difference in ion concentration between a high concentration electrolyte solution and a low concentration electrolyte solution, and the like .
  • the present invention is not limited thereto, and it is needless to say that various types of iontophoresis can be used.
  • An exosome kit for transcutaneous permeation enhancement of exosome comprises a first mask pack, a mask sheet or a patch which is applied or immersed with a composition containing an exosome as an active ingredient, And an iontophoresis device for applying a microcurrent to the skin of the skin.
  • an exosome kit for transcutaneous permeation enhancement of an exosome is characterized in that the first mask pack, the mask sheet or the patch is brought into contact with or attached to the skin of the mammal, By placing the iontophoresis device on a mask sheet or patch, the first mask pack, mask sheet or patch can be contacted or attached to be used in such a way that a microcurrent is applied to the skin of the mammal to which the composition is applied.
  • the composition may be used in combination with a conventionally used skin improving agent and / or moisturizer to the extent that the action thereof (improvement of skin condition, treatment of skin condition, skin disease, etc.) is not impaired.
  • the composition may be supported on or mixed with at least one of hydrogel, hyaluronic acid, hyaluronic acid salt (for example, sodium hyaluronate), or hyaluronic acid gel.
  • the type of the hydrogel is not limited, but it may preferably be a hydrogel obtained by dispersing a gelated polymer in a polyhydric alcohol.
  • the gel polymer is at least one selected from the group consisting of pluronic, purified agar, agarose, gellan gum, alginic acid, carrageenan, cacao gum, xanthan gum, galactomannan, glucomannan, pectin, cellulose, guar gum and locust bean gum
  • the polyhydric alcohol may be at least one selected from the group consisting of ethylene glycol, propylene glycol, 1,3-butylene glycol, isobutylene glycol, dipropylene glycol, sorbitol, xylitol and glycerin.
  • the iontophoresis device for enhancing transdermal permeation of exosome comprises a flexible battery, a lithium ion secondary battery, an alkaline battery, a dry battery, a mercury battery, a lithium battery, a nickel-cadmium A battery, and a reverse electrodialysis cell, or a second mask pack, a mask sheet, or a patch having the at least one battery mounted thereon.
  • the second mask pack, the mask sheet or the patch may be laminated on the first mask pack, the mask sheet or the patch.
  • At least one surface of the first mask pack, mask sheet or patch is coated with a hydrogel, hyaluronic acid, hyaluronate (for example, Sodium hyaluronate), or hyaluronic acid gel.
  • a hydrogel hyaluronic acid, hyaluronate (for example, Sodium hyaluronate), or hyaluronic acid gel.
  • the type of the hydrogel is not limited, but it may preferably be a hydrogel obtained by dispersing a gelated polymer in a polyhydric alcohol.
  • the gelled polymer and the polyhydric alcohol may be exemplified in the above description.
  • a micro current flows from the iontophoresis device so that the exosomes can effectively penetrate the epidermis and penetrate deeply into the skin.
  • the transdermal permeation enhancement method of exosome in one embodiment of the present invention can be carried out using the exosome kit as described above.
  • a method for enhancing transdermal permeability of an exosome comprises the steps of: (a) applying a first mask pack, a mask sheet or patch on which a composition comprising an exosome as an active ingredient is applied or immersed, (B) placing the iontophoresis device on the first mask pack, mask sheet or patch, and (c) contacting the first mask pack, mask sheet or patch with the first mask pack, (D) delivering the exosomes through the microcurrent to the interior of the mammalian skin.
  • At least one surface of the first mask pack, mask sheet or patch is coated with a hydrogel, hyaluronic acid, hyaluronic acid salt (for example, sodium hyaluronate ), Or hyaluronic acid gel.
  • a hydrogel hyaluronic acid, hyaluronic acid salt (for example, sodium hyaluronate ), Or hyaluronic acid gel.
  • the type of the hydrogel is not limited, but it may preferably be a hydrogel obtained by dispersing a gelated polymer in a polyhydric alcohol.
  • the gelled polymer and the polyhydric alcohol may be exemplified in the above description.
  • the iontophoresis device may be a flexible battery, a lithium ion secondary battery, an alkaline battery, a dry battery, a mercury battery, a lithium battery, a nickel- An electrodialysis cell, or a second mask pack, a mask sheet, or a patch with the at least one battery mounted thereon.
  • the step (b) may be performed by laminating the second mask pack, the mask sheet or the patch on the first mask pack, the mask sheet or the patch.
  • the exosome can be obtained by performing the following steps: (a) adding trehalose to the biological solution, (b) adding the trehalose (C) separating the exosomes from the filtered biological solution using TFF (Tangential Flow Filtration), and (d) removing the buffer used for desalting and buffer exchange (diafiltration). , Adding trehalose to the solution, and performing desalting and buffer exchange (diafiltration) on the separated exosome using TFF (Tangential Flow Filtration) using the buffer solution to which the trehalose is added .
  • TFF Tangential Flow Filtration
  • exosomes having a uniform particle size distribution and high purity can be effectively obtained (FIGS. 6A to 6C) 6E).
  • trehalose in the pre-filtration step (step (b) before exocase separation by TFF) and desalting by TFF and buffer exchange step (step (d)) after separation of exosome, a high purity particle size distribution Uniform exosome can be obtained with high yield.
  • Trehalose provides the ability to efficiently isolate exosomes for impurities such as cellular debris, waste products, proteins and macromolecules.
  • the desalting and buffer exchange can be performed continuously or intermittently. Desalting and buffer exchange can be carried out using a buffer solution having a volume of at least 4 times, preferably 6 times to 10 times, more preferably 12 times the starting volume.
  • a molecular weight cutoff (MWCO) 100,000 Da (Dalton), 300,000 Da, 500,000 Da or 750,000 Da TFF filter or 0.05 ⁇ ⁇ TFF filter can be used.
  • the step (c) may further include a step of concentrating the solution to a volume of 1/100 to 1/25 using TFF (Tangential Flow Filtration).
  • the biological solution may be a stem cell culture solution.
  • the type of the stem cell is not limited, but may be a mesenchymal stem cell, for example, a fat, a bone marrow, an umbilical cord or a cord blood-derived stem cell, more preferably a fat-derived stem cell.
  • the type of the adipose-derived stem cell is not limited as long as it does not cause a risk of infection by a pathogen and does not cause an immune rejection reaction, but it may be preferably a human adipose-derived stem cell.
  • the exosome used in the present invention is not limited to the exosome obtained according to the separation method described above, and it is needless to say that various exosomes which are used in the art or can be used in the future can be used. It is to be understood that the exosome isolated according to the above separation method should be understood as an example of exosome that can be used in the composition of the present invention, and the present invention is not limited thereto.
  • conditioning of the skin means improvement of the condition of the skin and / or prevention of the condition of the skin
  • improvement of the condition of the skin means visual and / Or a tactile perceptible positive change.
  • skin condition improvement may include skin regeneration, wrinkle removal or improvement, whitening, scar removal, skin texture improvement, redness reduction, moistening or smoothing of the condition of the skin, Rubbing can be an improvement of the deteriorated skin condition.
  • a method for enhancing transdermal permeation of an exosome comprising the steps of: (a) performing a transcutaneous permeation enhancing method of exosome as described above; and (b) The method comprising the steps of:
  • the present invention has the effect of facilitating skin application of exosomes that can function as functional cosmetics or medicines, and penetrating deeply into the skin while allowing the exosomes to pass the skin barrier effectively. Therefore, the present invention can be applied to a skin beauty which improves a poor skin condition or restores to a normal state, and can exhibit an excellent effect. In addition, the present invention provides an effect of treating a skin disease which alleviates, It is possible to exhibit an excellent effect.
  • FIG. 1 is a flow chart illustrating a process for separating and purifying exosomes in a method for producing exosomes from a biological solution according to one embodiment of the present invention.
  • FIG. 2 is a graph showing a result of measuring a relative amount of protein contained in a solution for each step of preparing a biological solution, for example, a stem cell culture solution, according to an embodiment of the present invention .
  • the ratio of the total amount of protein in each step was expressed by the relative ratio of total protein amount to the total biological solution.
  • the experimental results show the results obtained in two different batches, respectively.
  • FIG. 3 shows the results of measuring the productivity and purity of exosomes obtained according to one embodiment of the present invention.
  • the productivity of exosomes was calculated as "the number of particles of exosome obtained per mL of biological solution, eg, a stem cell culture (CM),” and the purity of exosome was calculated as "the number of exosomes per microgram of protein contained in the final fraction Quot; number of particles "
  • the experimental results show the results obtained in five different batches.
  • 4A to 4E show results of physical property analysis of exosomes obtained according to one embodiment of the present invention.
  • 4A shows particle size distribution and number of particles by TRPS (tunable resistive pulse sensing) analysis.
  • Figure 4B shows particle size distribution and number of particles by NTA (nanoparticle tracking analysis) analysis.
  • 4C shows particle images by transmission electron microscopy (TEM) according to magnification.
  • 4D shows the Western blot results of exosomes obtained according to one embodiment of the present invention.
  • &Quot; Figure 4E shows flow cytometric analysis results for CD63 and CD81 in marker assays for exosomes obtained according to one embodiment of the present invention.
  • Figures 5A-5C show the results of NTA analysis on particle size distribution showing that a uniform and high-purity exosome is obtained with trehalose addition. As the amount of trehalose added increases, a particle size distribution having a single peak can be obtained.
  • 6A to 6C show results of NTA analysis showing particle size distribution according to whether trehalose was added in the process of producing exosome according to one embodiment of the present invention.
  • 6A shows a case where trehalose is added throughout the production process
  • Fig. 6B shows a case where trehalose is added after the cell culture solution is stored in a frozen state
  • the results are shown without adding os.
  • 6D shows the results of comparing relative productivity and relative concentration of exosomes isolated by the methods of FIGS. 6A to 6C.
  • 6E shows the mean size of exosomes isolated by the methods of FIGS. 6A to 6C.
  • FIG. 7 shows the result of confirming the absence of cytotoxicity after treatment of exosome according to one embodiment of the present invention with HS68 cells of human skin fibroblasts.
  • FIG. 8 shows fluorescence intensity measurement results of exosomes stained with PKH67.
  • Fig. 9 is an image of a fluorescence microscope confirming the degree of transfer of fluorescent stained exosomes into pig skin tissue.
  • FIG. 10 is a confocal fluorescence microscope image showing the degree of fluorescence-stained exosomes transferred into the mouse skin tissue.
  • FIG. 11 is a graph showing a comparison of the total fluorescence intensity obtained by measuring the fluorescence intensity on each image in Fig.
  • FIG. 12 is a perspective view showing a configuration of the exosomes kit 100 according to one embodiment of the present invention.
  • FIG. 13 is a photograph showing the skin condition before and after treatment of human skin with the exosomal kit of the present invention.
  • FIG. 14 is a graph showing the results of application of iontophoresis for applying a microcurrent to a skin (affected part) coated with the composition after applying the composition containing exosome used in the exosome kit of the present invention to human skin As a result, it is a photograph showing that erythema of the skin (lesion) is remarkably improved.
  • FIG. 15 is a photograph showing the skin condition before and after the treatment of the exosome kit of the present invention on the face of a person suffering from acne;
  • 16A to 16I show the results of improving skin wrinkles, improving skin elasticity, skin moisturizing and dermal density when the exosome kit of the present invention is applied to human skin.
  • HS68 cells a human dermal fibroblast, were purchased from ATCC and cultured in RPMI 1640 supplemented with 10% fetal bovine serum (purchased from ThermoFisher Scientific) and 1% antibiotic-antimycotics (purchased from ThermoFisher Scientific) The cells were subcultured in DMEM (purchased from ThermoFisher Scientific) medium containing 5% CO 2 at 37 ° C.
  • adipose-derived stem cells were cultured at 5% CO 2 and 37 ° C. Then, the cells were washed with a phosphate-buffered saline (purchased from ThermoFisher Scientific), replaced with serum-free, non-phenol red medium, cultured for 1 to 10 days, and the supernatant .
  • a phosphate-buffered saline purchased from ThermoFisher Scientific
  • Trehalose was added to the culture medium in an amount of 2% by weight in order to obtain an exosome having a uniform particle size distribution and high purity in the process of separating exosome.
  • the culture was filtered with a 0.22 ⁇ m filter to remove impurities such as cellular debris, waste products and large particles.
  • the filtered cultures were immediately separated to isolate exosomes.
  • the filtered culture was stored in a refrigerator (image below 10 ° C) and used for exosome isolation.
  • the filtered culture was frozen in an ultra-low temperature freezer at -60 ° C or lower, and thawed, followed by exosome isolation. Then, the exosomes were separated from the culture medium by using Tangential Flow Filtration (TFF).
  • TMF Tangential Flow Filtration
  • Example 1 a TFF (Tangential Flow Filtration) method was used for separating, concentrating, desalting and diafiltration exosomes from a culture filtrated with a 0.22 ⁇ m filter.
  • a cartridge filter also called a hollow fiber filter (purchased from GE Healthcare) or a cassette filter (purchased from Pall or Sartorius or Merck Millipore) was used.
  • the TFF filter can be selected by a variety of molecular weight cutoffs (MWCO). Exosomes were selectively isolated and concentrated by selected MWCOs, and particles, proteins, lipids, nucleic acids, low molecular weight compounds, etc. smaller than MWCO were removed.
  • MWCO molecular weight cutoffs
  • TFF filters of MWCO 100,000 Da (Dalton), 300,000 Da, or 500,000 Da were used.
  • the culture medium was concentrated to a volume of about 1/100 to 1/25 using the TFF method, and substances smaller than MWCO were removed to separate the exosomes.
  • Separated and concentrated exosomal solutions were further desalted and buffered (diafiltration) using the TFF method.
  • the desalination and buffer exchange are performed by continuous diafiltration or discontinuous diafiltration, and at least 4 times, preferably 6 times to 10 times or more, more preferably, Was performed using a buffer solution having a volume of 12 times or more.
  • 2% by weight of trehalose dissolved in PBS was added to obtain an exosome having a uniform particle size distribution and high purity.
  • 6A to 6E show the effect of obtaining an exosome having a high purity and a uniform particle size distribution with trehalose treatment at a high yield.
  • the amount of protein in the fractions of the isolated exosome, culture medium, and TFF separation process was measured using BCA colorimetry (purchased from ThermoFisher Scientific) or FluoroProfile fluorescence (purchased from Sigma).
  • BCA colorimetry purchased from ThermoFisher Scientific
  • FluoroProfile fluorescence purchased from Sigma.
  • the extent to which the exosome was isolated and concentrated by the TFF method of the present invention and the removal of proteins, lipids, nucleic acids, and low-molecular compounds was monitored by a protein determination method, and the results are shown in FIG. As a result, it was found that the protein present in the culture solution was very effectively removed by the TFF method of one embodiment of the present invention.
  • FIG. 3 shows the results of comparing the productivity and purity in five independent batches when isolating exosomes by the TFF method of one embodiment of the present invention. As a result of analyzing the results obtained from the five independent batches, it was confirmed that the exosome can be separated very stably by the TFF method of one embodiment of the present invention.
  • 5A to 5C show results of NTA analysis of the size distribution of exosome according to whether or not trehalose was added after the exosome was separated by the TFF method.
  • concentration of trehalose was increased to 0% by weight, 1% by weight and 2% by weight (from top to bottom of FIGS. 5A to 5C) and repeated three times.
  • particles having a size of 300 nm or more were identified, while particles having a size of 300 nm or more were reduced by increasing the amount of trehalose, and the size distribution of the exosome was uniformized .
  • FIG. 4D shows the presence of CD9, CD63, CD81 and TSG101 markers as a result of performing Western blotting on isolated exosomes according to the method of one embodiment of the present invention.
  • Anti-CD9 purchased from Abcam
  • anti-CD63 purchasedd from System Biosciences
  • anti-CD81 purchasedd from System Biosciences
  • anti-TSG101 purchasedd from Abcam
  • FIG. 4E shows the presence of CD63 and CD81 markers as a result of analysis using a flow cytometer on exosomes isolated according to the method of one embodiment of the present invention.
  • Human CD63 isolation / detection kit purchased from ThermoFisher Scientific
  • PE-mouse anti-human CD63 PE-Mouse anti markers were stained using the PE-mouse CD63 (purchased from BD) and PE-mouse anti-human CD81 (purchased from BD), and analyzed using a flow cytometer (ACEA Biosciences) Respectively.
  • exosome used in the present invention is not limited to the exosome of the above-mentioned embodiments, and it is needless to say that various exosomes which are used in the art or can be used in the future can be used. It should be understood that the exosome isolated according to the above embodiments is an example of exosome that can be used in the present invention, and the present invention is not limited thereto.
  • exosomes were treated by concentration to the cells and the proliferation rate of the cells was confirmed.
  • HS68 cells were suspended in DMEM containing 10% FBS, and the cells were mixed with 80 ⁇ 90% confluency and cultured in a 5% CO 2 incubator at 37 ° C for 24 hours. After 24 hours, the culture solution was removed, and the cell survival rate was evaluated by culturing the exosome prepared in Example 2 for each concentration and culturing for 24 to 72 hours.
  • WST-1 reagent purchased from Takara
  • MTT reagent purchased from Sigma
  • CellTiter-Glo reagent purchased from Promega
  • Aramar Blue reagent alamarBlue reagent purchased from ThermoFisher Scientific
  • a microplate reader purchased from Molecular Devices
  • the comparison group was based on the number of cells cultured in the normal cell culture medium not treated with exosome, and it was confirmed that no cytosoxicity by exosome was observed within the tested concentration range (FIG. 7).
  • PKH67 dye (purchased from Sigma) was used to prepare fluorescently stained exosomes. 1 mM PKH67 was diluted in Diluent C (purchased from Sigma) to prepare a 10 ⁇ M PKH67 solution, mixed with an appropriate concentration of exosome solution, and allowed to react at room temperature for 10 minutes in the absence of light. After the reaction, an MW3000 spin column (purchased from ThermoFisher Scientific) was used to remove the remaining PKH67 dye from the exosomes stained with PKH67 (hereinafter abbreviated as "PKH-exosomes").
  • Example 6 Skin permeability test of exosome used in the exosome kit of the present invention
  • PKH-exosomes were dispersed in phosphate buffer (PBS) at a proper concentration, for example, 1 ⁇ 10 5 to 1 ⁇ 10 9 particles / mL, and then applied to the outer surface of the pig skin.
  • PBS phosphate buffer
  • the nonwoven fabric was covered and allowed to react for an appropriate period of time, for example, 30 minutes to 1 hour to allow the PKH-exosomes to be delivered to the subcutaneous tissues.
  • the PKH-exosomes were applied to the outer surface of the pig skin and covered with the nonwoven fabric, followed by flowing a microcurrent for a predetermined time, for example, 30 minutes to 1 hour.
  • the pig skin tissue was immersed in a 3.7% formaldehyde solution, reacted overnight, and washed three times for 5 minutes each with phosphate buffered saline.
  • the washed pig skin tissue was immersed in a 30% sucrose solution and treated with OCT compound.
  • sections were prepared on a longitudinal section using a microtome, and tissue sections were placed on a slide glass.
  • the preparation of the tissue section may be carried out before the tissue is fixed with formaldehyde solution. Fluorescence detected from PKH-exosomes in tissue sections was observed using fluorescence microscopy.
  • PKH-exosomes transmitted to the subcutaneous tissue through the epidermis of the pig skin tissue were identified.
  • the exosomes were more effectively passed through the epidermis, deeply transferred into the skin tissue, and more effectively absorbed to the skin when active current was delivered (active transdermal delivery) (see FIG. 9).
  • the exosomes used in the exosome kit of the present invention can effectively pass through the epidermis and be deeply transferred into the skin tissue, and can be effectively absorbed to the skin.
  • PKH-exosomes were dispersed in phosphate buffer (PBS) at a proper concentration, for example, 1 ⁇ 10 5 to 1 ⁇ 10 9 particles / mL, and then applied to the outside of the skin tissue of the mice.
  • PBS phosphate buffer
  • the nonwoven fabric was pre-positioned outside the mouse skin tissue, and PKH-exosomes were injected between the nonwoven fabric and skin tissue. Then, the reaction was carried out for 30 minutes to 1 hour.
  • the PKH-exosomes were injected between the nonwoven fabric and the skin tissue, and the microcurrent was flowed for a predetermined time, for example, 30 minutes to 1 hour. After completion of the reaction, the PKH-exosomes transferred into the skin tissue were confirmed immediately using a confocal fluorescence microscope (Leica, SP8X) or the skin tissue and the PKH-exosomal solution were further reacted for 1 hour to 6 hours , And PKH-exosomes were identified by confocal fluorescence microscopy. As a result, it was confirmed that the exosomes were more efficiently passed through the epidermis and deeply transferred into the skin tissue when the microcurrent was passed (active transdermal delivery), and more effectively absorbed to the skin (Figs. 10 and 11) .
  • the exosome kit 100 includes a liquid immersion sheet mask 10 (a white sheet mask coated or immersed with a composition containing the exosome prepared in Example 2) And a percutaneous permeation promoting sheet mask (silver sheet mask) 20 which is an iontophoresis device.
  • Exosome 12 is coated or deposited on the surface or inside of liquid immersion sheet mask 10.
  • a battery 22 for generating a microcurrent is mounted on the percutaneous permeation enhancing sheet mask 20 and a conductive pattern 24 electrically connected to the cell 22 is attached to the percutaneous permeation enhancing sheet mask 20, So that the micro current generated from the battery 22 can be uniformly transmitted to the entirety of the percutaneous permeation enhancing sheet mask 20.
  • the conductive pattern 24 is formed by printing or plating a conductive metal such as gold, silver, or copper.
  • exosome kit 100 having the above-described structure is applied once to the face of a person having a flushing symptom and a severe skin trouble (hereinafter referred to as "case 1") to alleviate or alleviate skin troubles and flushing symptoms .
  • case 1 a severe skin trouble
  • exosome kit 100 of the present invention was applied once to the face of a person with flushing symptom (hereinafter referred to as " case 2 ") to evaluate whether the overall skin tone and flushing symptom were improved.
  • a liquid immersion sheet mask 10 containing 5 ⁇ 10 5 exosomes 12 was uniformly adhered to the entire faces of the cases 1 and 2 by aligning them with the eyes and mouth parts and then a microcurrent of about 0.3 to 0.4 mA
  • An activated percutaneous permeation enhancing sheet mask 20 which was activated to flow was laminated on the liquid immersion sheet mask 10 and used for about 25 to 30 minutes.
  • the exosomatic kit 100 of the present invention which comprises the liquid immersion sheet mask 10 on which the composition containing the exosome as an active ingredient is immersed and the percutaneous permeation promoting sheet mask (iontophoresis device) 20, It can be said that there is a skin-beauty effect or a skin disease treatment effect that prevents, suppresses, alleviates or improves symptoms and skin troubles.
  • a composition (exosome suspension) containing the exosome prepared in Example 2 at a concentration of 0.29 x 10 8 particles / mL was applied to the affected part (hand, neck, arm, etc.) of three severe atopic patients complaining of itching
  • An iontophoresis was performed using a iontophoresis device (IONZYME DF MACHINE) (purchased from Environ) to flow a micro current of 0.4 mA for 20 minutes to the affected part of the composition.
  • IONZYME DF MACHINE purchased from Environ
  • the exosome kit 100 of the present invention can be said to have a skin-beauty effect or a skin disease-treating effect for preventing, suppressing, alleviating or alleviating itching and erythema of the skin as confirmed by the clinical test as described above.
  • Example 8 Treatment of exosome kit on facial surface of subjects with acne symptoms
  • Example 7 The "exosome kit" described in Example 7 was applied to the face of a person suffering from acne symptomatically once every four days to evaluate whether acne symptoms and related skin troubles were alleviated or improved.
  • a liquid immersion sheet mask 10 containing 5 ⁇ 10 5 exosomes 12 was applied to the entire face of a person suffering from acne symptomatically by aligning them with the eyes and the mouth and then a micro current of about 0.3 to 0.4 mA
  • An activated percutaneous permeation enhancing sheet mask 20 is laminated on the liquid immersion sheet mask 10 and used for about 25 to 30 minutes.
  • the exosome kit 100 of the present invention can be said to have a skin-beauty effect or a skin disease treatment effect for preventing, suppressing, alleviating or alleviating acne symptoms and skin troubles associated therewith.
  • Example 9 Test for improving wrinkles, improving elasticity, moisturizing effect, etc. on human skin
  • the exosomes prepared in Example 2 were mixed at a concentration of 1 x 10 < 7 > (Translucent oil-in-water) and the exosome kit 100 of the present invention were used once every three days in the evening to test skin cosmetic effects such as skin wrinkles, skin elasticity improvement, moisturizing and skin regeneration effects.
  • the liquid immersion sheet mask 10 was brought into close contact with the eyes and the mouth, and the entire surface of the face was evenly adhered. Then, an activated percutaneous permeation- The mask 20 was laminated on the liquid immersion sheet mask 10 and used for about 25 to 30 minutes.
  • the skin moisturizing effect was measured by a moisture meter (Corneometer CM825) (Courage-Khazaka eletronic GmbH, Germany) And skin elasticity was evaluated by measuring the sound pressure with CM580 (Cutometer CM580) (Courage + Khazaka electronic GmbH, Germany).
  • CM580 Cutometer CM580
  • Dermal density improvement was assessed by analyzing the density (%) after the skin scanner ultrasonography at the eye.
  • the test subjects were tested for the test items after the use of the exosomatic kit 100 of the present invention in five steps of excellent (4), good (3), normal (2), poor (1) and very poor (0) And responded to the questionnaire.
  • the questionnaire survey the mean value and the standard deviation of the questionnaire and the percentage of the subjects to the answer were obtained for each test item.
  • 100% of the test subjects were evaluated as above all the test items.
  • the subjects were asked to answer the questionnaire about feeling of use of the exosome kit (100) of the present invention.
  • the exosome kit 100 of the present invention is excellent in skin wrinkles, skin elasticity, skin regeneration and skin moisturizing effects, as confirmed by the above-mentioned clinical tests.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Epidemiology (AREA)
  • Chemical & Material Sciences (AREA)
  • Birds (AREA)
  • Cell Biology (AREA)
  • Medicinal Chemistry (AREA)
  • Developmental Biology & Embryology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Immunology (AREA)
  • Zoology (AREA)
  • Biomedical Technology (AREA)
  • Biotechnology (AREA)
  • Virology (AREA)
  • Dermatology (AREA)
  • Engineering & Computer Science (AREA)
  • Hematology (AREA)
  • Dispersion Chemistry (AREA)
  • Medicinal Preparation (AREA)

Abstract

La présente invention concerne un kit d'exosomes pour l'amélioration de la perméation transdermique d'exosomes, comprenant un dispositif d'iontophorèse, un conditionnement de masque et un masque revêtu d'une composition contenant des exosomes en tant que principe actif ou sur lequel une telle composition est déposée. La présente invention facilite l'application sur la peau d'exosomes, qui peuvent fonctionner en tant que produit cosmétique fonctionnel ou quasi-médicament et permet aux exosomes de pénétrer profondément dans la peau en traversant efficacement la barrière cutanée. Par conséquent, la présente invention peut présenter d'excellents effets lorsqu'elle est utilisée pour traiter la peau afin d'atténuer des lésions cutanées ou pour faire retrouver à la peau son état normal, et peut présenter d'excellents effets sur le traitement des maladies de peau permettant une atténuation ou un soulagement des lésions cutanées ou le retour de la peau à son état normal.
PCT/KR2018/012549 2017-10-31 2018-10-23 Kit d'exosomes et procédé d'amélioration de la perméation transdermique d'exosomes l'utilisant Ceased WO2019088545A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
KR20170143358 2017-10-31
KR10-2017-0143358 2017-10-31

Publications (1)

Publication Number Publication Date
WO2019088545A1 true WO2019088545A1 (fr) 2019-05-09

Family

ID=66333242

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/KR2018/012549 Ceased WO2019088545A1 (fr) 2017-10-31 2018-10-23 Kit d'exosomes et procédé d'amélioration de la perméation transdermique d'exosomes l'utilisant

Country Status (2)

Country Link
KR (1) KR102320845B1 (fr)
WO (1) WO2019088545A1 (fr)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20240207143A1 (en) * 2021-04-29 2024-06-27 Reza Mollaaghababa Articles for streamlined skin rejuvenation and wound healing using exosomes and prp

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR102347311B1 (ko) * 2019-03-11 2022-01-06 바이오센서연구소 주식회사 역전기투석과 산화-환원 반응을 이용한 장치 및 그를 사용하여 약물을 전달하는 방법
KR102565365B1 (ko) * 2022-01-24 2023-08-09 주식회사 엑소코바이오 캣닢 유래의 엑소좀을 유효성분으로 포함하는 새로운 조성물

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR100628946B1 (ko) * 2005-04-04 2006-09-27 주식회사 엘지생활건강 미세 전류를 이용한 마스크 팩 세트
KR20160086253A (ko) * 2015-01-08 2016-07-19 한양대학교 에리카산학협력단 줄기세포로부터 추출된 엑소좀을 함유하는 피부 미백, 주름개선 또는 재생용 화장료 조성물
KR20160116491A (ko) * 2015-03-30 2016-10-10 조정호 스마트폰을 이용하는 패턴화된 전도성 탄소필름 지지체를 구비한 전류마스크팩 장치
KR20170044999A (ko) * 2015-10-16 2017-04-26 (주)프로스테믹스 줄기세포 유래 엑소좀을 포함하는 상처 치료, 피부 개선 및 탈모 방지 또는 치료용 조성물 및 그 제조방법
KR20170104821A (ko) * 2016-03-08 2017-09-18 (주)아이보손 마스크 팩용 이온도입장치

Family Cites Families (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP2254586B1 (fr) * 2008-02-22 2015-04-08 Agency For Science, Technology And Research (A*star) Particules de cellules souches mésenchymateuses
KR20090111269A (ko) * 2009-02-10 2009-10-26 (주)프로스테믹스 아토피성 피부염의 예방 또는 치료용 조성물
KR101443180B1 (ko) 2012-11-27 2014-09-22 한국콜마주식회사 신규한 경피흡수용 약물전달체, 그를 이용한 탈모방지 외용제 조성물 및 화장료
US20160089535A1 (en) * 2014-09-29 2016-03-31 Elc Management Llc Targeted And Individualized Delivery Of Skincare Treatments With Micro-Current In A Mask Or Patch Form
WO2016072821A1 (fr) 2014-11-07 2016-05-12 한양대학교 에리카산학협력단 Composition pour l'induction de la différenciation d'adipocyte contenant un exosome dérivé de cellules souches, la régénération de tissus adipeux, et la décoloration de la peau ou l'atténuation des rides
KR101784050B1 (ko) 2014-12-23 2017-10-10 영남대학교 산학협력단 엑소솜을 유효성분으로 함유하는 피부 노화 예방 또는 치료용 약학조성물
KR101686064B1 (ko) 2015-09-21 2016-12-13 (주)프로스테믹스 면역 억제 및 항염증성 조성물
WO2017119520A1 (fr) * 2016-01-05 2017-07-13 바이오센서연구소 주식회사 Masque facial utilisant l'électrodialyse inverse et kit le comprenant

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR100628946B1 (ko) * 2005-04-04 2006-09-27 주식회사 엘지생활건강 미세 전류를 이용한 마스크 팩 세트
KR20160086253A (ko) * 2015-01-08 2016-07-19 한양대학교 에리카산학협력단 줄기세포로부터 추출된 엑소좀을 함유하는 피부 미백, 주름개선 또는 재생용 화장료 조성물
KR20160116491A (ko) * 2015-03-30 2016-10-10 조정호 스마트폰을 이용하는 패턴화된 전도성 탄소필름 지지체를 구비한 전류마스크팩 장치
KR20170044999A (ko) * 2015-10-16 2017-04-26 (주)프로스테믹스 줄기세포 유래 엑소좀을 포함하는 상처 치료, 피부 개선 및 탈모 방지 또는 치료용 조성물 및 그 제조방법
KR20170104821A (ko) * 2016-03-08 2017-09-18 (주)아이보손 마스크 팩용 이온도입장치

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20240207143A1 (en) * 2021-04-29 2024-06-27 Reza Mollaaghababa Articles for streamlined skin rejuvenation and wound healing using exosomes and prp

Also Published As

Publication number Publication date
KR20190049474A (ko) 2019-05-09
KR102320845B1 (ko) 2021-11-02

Similar Documents

Publication Publication Date Title
KR102163806B1 (ko) 줄기세포 유래의 엑소좀을 유효성분으로 포함하는 피지분비 감소용 조성물
JP7037205B2 (ja) 幹細胞由来のエキソソームを有効成分として含む組成物のかゆみの抑制又は改善の用途
KR102755006B1 (ko) 줄기세포 유래의 엑소좀을 유효성분으로 포함하는 민감성 피부의 예방, 억제, 완화 또는 개선용 화장료 조성물
KR102646145B1 (ko) 줄기세포 유래의 엑소좀을 유효성분으로 포함하는 모공 축소용 조성물
WO2019168281A1 (fr) Procédé pour soins de la peau par exposition de la peau à un faisceau laser et traitement par exosomes dérivés de cellules souches
CN110123838B (zh) 负载白藜芦醇的人多能干细胞外泌体及其制备方法与用途
KR20190003399A (ko) 줄기세포 유래의 엑소좀을 유효성분으로 포함하는 조성물의 피부 섬유증 개선 용도
WO2019088545A1 (fr) Kit d'exosomes et procédé d'amélioration de la perméation transdermique d'exosomes l'utilisant
WO2019112238A2 (fr) Composition cosmétique pour hydrater la peau comprenant un exosome dérivé de cellules souches en tant qu'ingrédient actif
WO2019182299A1 (fr) Procédé de soin de la peau utilisant le rayonnement ipl sur la peau et traitement d'exosome dérivé de cellules souches en combinaison
KR20190045829A (ko) 엑소좀의 경피 투과 증진 방법 및 이의 응용
KR20190069291A (ko) 줄기세포 유래의 엑소좀을 유효성분으로 포함하는 피부 보습용 화장료 조성물
WO2021125558A1 (fr) Composition pour le noircissement des cheveux, la prévention du blanchiment des cheveux, ou la teinture des cheveux, comprenant un exosome dérivé de cellules souches en tant que principe actif
KR20240037208A (ko) 줄기세포 유래의 엑소좀을 유효성분으로 포함하는 안면 홍조 개선용 조성물
KR102689564B1 (ko) 피부 필링과 줄기세포 유래의 엑소좀 처리를 병용한 피부 미용방법
KR101964991B1 (ko) 줄기세포 유래의 엑소좀을 포함하는 엑소좀 키트의 새로운 용도
KR20190114740A (ko) 피부에 대한 ｒｆ 에너지 인가와 줄기세포 유래의 엑소좀 처리를 병용한 피부 미용방법
KR20200038672A (ko) 피부에 대한 플라즈마 처리와 엑소좀 처리를 병용한 피부 상태의 개선 방법
WO2019190093A1 (fr) Procédé de soin de la peau utilisant, en combinaison, l'application d'énergie rf sur la peau et le traitement par exosomes dérivés de cellules souches sur la peau
WO2019231133A1 (fr) Composition pour réduire les pores, comprenant des exosomes dérivés de cellules souches en tant que principe actif
KR20190027720A (ko) 줄기세포 유래의 엑소좀을 포함하는 엑소좀 키트의 새로운 용도

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 18874650

Country of ref document: EP

Kind code of ref document: A1

ENP Entry into the national phase

Ref document number: 2020600104

Country of ref document: JP

Kind code of ref document: A

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 18874650

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: JP