WO2019168281A1 - Procédé pour soins de la peau par exposition de la peau à un faisceau laser et traitement par exosomes dérivés de cellules souches - Google Patents

Procédé pour soins de la peau par exposition de la peau à un faisceau laser et traitement par exosomes dérivés de cellules souches Download PDF

Info

Publication number
WO2019168281A1
WO2019168281A1 PCT/KR2019/001677 KR2019001677W WO2019168281A1 WO 2019168281 A1 WO2019168281 A1 WO 2019168281A1 KR 2019001677 W KR2019001677 W KR 2019001677W WO 2019168281 A1 WO2019168281 A1 WO 2019168281A1
Authority
WO
WIPO (PCT)
Prior art keywords
skin
laser beam
exosomes
beam irradiation
composition
Prior art date
Application number
PCT/KR2019/001677
Other languages
English (en)
Korean (ko)
Inventor
조병성
이용원
Original Assignee
주식회사 엑소코바이오
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 주식회사 엑소코바이오 filed Critical 주식회사 엑소코바이오
Publication of WO2019168281A1 publication Critical patent/WO2019168281A1/fr

Links

Images

Classifications

    • 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
    • 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—Liposomes
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
    • A61M37/00—Other apparatus for introducing media into the body; Percutany, i.e. introducing medicines into the body by diffusion through the skin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61N—ELECTROTHERAPY; MAGNETOTHERAPY; RADIATION THERAPY; ULTRASOUND THERAPY
    • A61N1/00—Electrotherapy; Circuits therefor
    • A61N1/02—Details
    • A61N1/04—Electrodes
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61N—ELECTROTHERAPY; MAGNETOTHERAPY; RADIATION THERAPY; ULTRASOUND THERAPY
    • A61N1/00—Electrotherapy; Circuits therefor
    • A61N1/02—Details
    • A61N1/04—Electrodes
    • A61N1/0404—Electrodes for external use
    • A61N1/0408—Use-related aspects
    • A61N1/0428—Specially adapted for iontophoresis, e.g. AC, DC or including drug reservoirs
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61N—ELECTROTHERAPY; MAGNETOTHERAPY; RADIATION THERAPY; ULTRASOUND THERAPY
    • A61N1/00—Electrotherapy; Circuits therefor
    • A61N1/18—Applying electric currents by contact electrodes
    • A61N1/32—Applying electric currents by contact electrodes alternating or intermittent currents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61N—ELECTROTHERAPY; MAGNETOTHERAPY; RADIATION THERAPY; ULTRASOUND THERAPY
    • A61N1/00—Electrotherapy; Circuits therefor
    • A61N1/18—Applying electric currents by contact electrodes
    • A61N1/32—Applying electric currents by contact electrodes alternating or intermittent currents
    • A61N1/325—Applying electric currents by contact electrodes alternating or intermittent currents for iontophoresis, i.e. transfer of media in ionic state by an electromotoric force into the body
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61N—ELECTROTHERAPY; MAGNETOTHERAPY; RADIATION THERAPY; ULTRASOUND THERAPY
    • A61N5/00—Radiation therapy
    • A61N5/06—Radiation therapy using light
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61N—ELECTROTHERAPY; MAGNETOTHERAPY; RADIATION THERAPY; ULTRASOUND THERAPY
    • A61N5/00—Radiation therapy
    • A61N5/06—Radiation therapy using light
    • A61N5/0613—Apparatus adapted for a specific treatment
    • A61N5/0616—Skin treatment other than tanning
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61N—ELECTROTHERAPY; MAGNETOTHERAPY; RADIATION THERAPY; ULTRASOUND THERAPY
    • A61N5/00—Radiation therapy
    • A61N5/06—Radiation therapy using light
    • A61N5/067—Radiation therapy using light using laser light
    • 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
    • A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
    • A61M37/00—Other apparatus for introducing media into the body; Percutany, i.e. introducing medicines into the body by diffusion through the skin
    • A61M2037/0007—Other apparatus for introducing media into the body; Percutany, i.e. introducing medicines into the body by diffusion through the skin having means for enhancing the permeation of substances through the epidermis, e.g. using suction or depression, electric or magnetic fields, sound waves or chemical agents

Definitions

  • the present invention relates to a skin care method using a combination of laser beam irradiation to the skin and exosomes derived from stem cells.
  • the present invention relates to a skin cosmetic method for reducing skin redness, such as side effects due to laser beam irradiation by treating the skin with exosomes derived from stem cells with laser beam irradiation.
  • Laser beam irradiation is known to increase collagen synthesis in the skin to help skin regeneration and restore skin elasticity to improve wrinkles, wounds, and the like.
  • a skin cosmetic method using laser beam irradiation utilizes a mechanism in which the skin is regenerated by activating the tissues in the surrounding area after thermal damage to the treatment site.
  • the laser beam When the laser beam is irradiated to the entire skin of the treatment area, it transmits light energy from the epidermis to the dermis layer to regenerate the skin widely.However, high energy transfer causes severe pain in the treatment area and causes redness or swelling of the skin after the procedure. .
  • too much laser heat damage due to operator error can render the regeneration mechanism impossible and deep scars may remain, and erythema, edema, hyperpigmentation and underpigmentation ( Side effects, such as hypopigmentation, can occur.
  • a method of irradiating a laser beam in the form of a spot is used, and in this case, a plurality of laser beam spots (micro laser beams) are formed using a micro lens array or a laser scanner.
  • the treatment method using the laser beam spot can improve the sequelae, since energy is delivered to a certain depth of penetration while forming a fine treatment area.
  • side effects still occur. Therefore, there is a need for research on other aspects to minimize side effects due to laser beam irradiation.
  • the cell secretome contains a variety of bioactive factors that control the behavior (behavior) of the cell, especially in the cell secretion 'exosomes (cell) having a signaling function between cells ', And its research on the composition and function is actively underway.
  • Extracellular vesicles are called cell membrane-derived vesicles, ectosomes, shedding vesicles, microparticles, exosomes, and the like, and in some cases, may be used separately from exosomes.
  • Exosomes are tens to hundreds of nanometers of endoplasmic reticulum consisting of a double phospholipid membrane identical to the structure of a cell membrane, and include proteins, nucleic acids (mRNA, miRNA, etc.) called exosome cargo.
  • Exosome cargo includes a wide range of signaling factors, which are known to be specific for cell types and differently regulated by the environment of the secretory cell.
  • Exosomes are intercellular signaling media secreted by cells, and the various cellular signals transmitted through them regulate cell behavior, including activation, growth, migration, differentiation, dedifferentiation, apoptosis, and necrosis of target cells. Known.
  • Exosomes contain specific genetic material and bioactive factors depending on the nature and state of the cells from which they are derived. Stem cell-derived exosomes, which proliferate, regulate cell behavior such as cell migration, proliferation and differentiation, and reflect stem cell characteristics related to tissue regeneration (Nature Review Immunology 2002 (2) 569-579).
  • the present inventors have been intensively researching new applications of stem cell-derived exosomes and incorporating them into medical or cosmetic technologies.
  • the present invention was completed by confirming the effect of reducing skin redness and the like caused by side effects.
  • An object of the present invention is to provide a skin care method using a combination of laser beam irradiation to the skin and exosomes derived from stem cells.
  • Another object of the present invention is to provide a skin cosmetic method for reducing skin redness, which is a side effect of laser beam irradiation, by treating the skin with exosomes derived from stem cells along with laser beam irradiation.
  • the present invention provides a method for irradiating a skin with a laser beam, before the laser beam irradiation, during the laser beam irradiation, or after the laser beam irradiation.
  • exosomes refers to vesicles of a size ranging from tens to hundreds of nanometers (preferably approximately 30 to 200 nm) consisting of a double phospholipid membrane identical to the structure of a cell membrane, provided that Particle size of exosomes may vary depending on the cell type, isolation method and measurement method) (Vasiliy S. Chernyshev et al., "Size and shape characterization of hydrated and desiccated exosomes", Anal Bioanal Chem, (2015) DOI 10.1007 / s00216-015-8535-3). Exosomes include proteins called exosome cargo (cargo), nucleic acids (mRNA, miRNA, etc.).
  • Exosome cargo includes a wide range of signaling factors, which are known to be specific for cell types and differently regulated by the environment of the secretory cell. Exosomes are intercellular signaling media secreted by cells, and the various cellular signals transmitted through them regulate cell behavior, including activation, growth, migration, differentiation, dedifferentiation, apoptosis, and necrosis of target cells. Known.
  • exosome has a nano-size vesicle structure secreted by stem cells and released into the extracellular space and a vesicle having a composition similar to exosomes (eg, exosomes- Pseudo vesicles).
  • the type of the stem cells is not limited, but as an example, which does not limit the present invention, preferably may be mesenchymal stem cells, for example, fat, bone marrow, umbilical cord or cord blood-derived stem cells, more preferably Fat-derived stem cells.
  • the type of the adipose derived stem cells is not limited as long as there is no risk of infection by the pathogen and does not cause an immune rejection reaction, but preferably, human adipose derived stem cells.
  • the stem cell-derived exosomes used in the present invention have an effect of reducing skin redness, which is a side effect of laser beam irradiation, and various stem cells that are used in the art or may be used in the future as long as they do not cause adverse effects on the human body.
  • skin redness which is a side effect of laser beam irradiation
  • various stem cells that are used in the art or may be used in the future as long as they do not cause adverse effects on the human body.
  • skin care refers to side effects due to laser beam irradiation, such as pain, redness or swelling of the skin after laser treatment, and / or pigmentation (eg hyperpigmentation and / or subpigmentation). ) Decreases, alleviates and / or improves the skin, and / or has a positive effect such as soothing the skin, shrinking the pores, and / or improving the skin brightness (eg, increasing the face radiance area).
  • the term “iontophoresis” refers to a method of allowing an ionized active ingredient to penetrate the skin with electrical repulsion by changing a skin's electrical environment by applying a potential difference by flowing a microcurrent to the skin to which the active material is applied. Means.
  • the iontophoresis used in one embodiment of the present invention is a method in which a current from an external power source flows into the electrode patch on the skin to introduce a microcurrent into the skin, and a battery is mounted on the electrode patch itself.
  • the manner in which the microcurrent is introduced the manner in which the microcurrent is introduced into the skin through a patch equipped with reversed electrodialysis means for generating a current through the difference in ion concentration between the high concentration electrolyte solution and the low concentration electrolyte solution.
  • the present invention is not limited thereto, and various methods of iontophoresis may be used.
  • the skin care method of an embodiment of the present invention comprises the steps of (a) irradiating a laser beam to the skin of a mammal, and (b) before or after the laser beam irradiation, to the skin of the mammal to be subjected to the laser beam irradiation Applying a composition comprising a stem cell-derived exosomes as an active ingredient.
  • the skin cosmetic method of an embodiment of the present invention can shorten downtime, which is a time taken for skin redness and swelling, which are side effects due to laser beam irradiation, to disappear.
  • the composition may be a cosmetic composition or a skin external preparation.
  • the cosmetic composition may be a cream or lotion.
  • the component usually used in cosmetics or external preparation for skin within the range that does not impair the effects of the present invention,
  • a moisturizer, antioxidant, oily component, ultraviolet absorber, emulsifier, surfactant, thickener, alcohol, powder component, colorant, aqueous component, water, various skin nutrients, etc. can be suitably blended as needed.
  • the composition may be used in combination with a conventional skin improver and / or moisturizers in addition to the exosomes derived from stem cells, as long as the action (reduction of side effects due to laser beam irradiation, etc.) is not impaired.
  • the exosomes derived from stem cells may be supported or mixed in at least one of a hydrogel, hyaluronic acid, a hyaluronic acid salt (for example, sodium hyaluronate), or a hyaluronic acid gel.
  • the type of the hydrogel is not limited, but may be preferably a hydrogel obtained by dispersing a gelling polymer in a polyhydric alcohol.
  • the gelling polymer is at least one selected from the group consisting of pluronic, purified agar, agarose, gellan gum, alginic acid, carrageenan, cassia 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 external preparation for skin and / or cosmetic composition of one embodiment of the present invention may include, for example, patches, mask packs, mask sheets, creams, tonics, ointments, suspensions, emulsions, pastes, lotions, gels, oils, packs, sprays, and aerosols. It can be applied to various forms such as, mist, foundation, powder and oil paper.
  • the external preparation for skin and / or cosmetic composition may be applied or deposited on at least one side of a patch, mask pack or mask sheet.
  • the external preparation for skin of one embodiment of the present invention is prepared with a cosmetic composition, for example, redness or swelling of the skin after laser treatment is reduced, pigmentation (for example, hyperpigmentation and / or underpigmentation) improvement, skin It may be used for the purpose of soothing, shrinking pores, improving skin brightness (eg, increasing face radiance, etc.), and the cosmetic formulation may be prepared in any formulation conventionally made in the art.
  • a cosmetic composition for example, redness or swelling of the skin after laser treatment is reduced, pigmentation (for example, hyperpigmentation and / or underpigmentation) improvement, skin
  • pigmentation for example, hyperpigmentation and / or underpigmentation
  • skin may be used for the purpose of soothing, shrinking pores, improving skin brightness (eg, increasing face radiance, etc.)
  • the cosmetic formulation may be prepared in any formulation conventionally made in the art.
  • patch for example, patch, mask pack, mask sheet, supple cosmetics, nourishing cosmetics, astringent cosmetics, nourishing cream, massage cream, eye cream, cleansing cream, essence, eye essence, cleansing lotion, cleansing foam, cleansing water, sunscreen, lipstick , Soaps, shampoos, surfactant-containing cleansing, bathing agents, body lotions, body creams, body oils, body essences, body cleaners, hair dyes, hair tonic and the like, but is not limited thereto.
  • the external preparation for skin and / or cosmetic composition of one embodiment of the present invention comprises ingredients conventionally used in external preparations for skin and / or cosmetics, such as conventional adjuvants such as antioxidants, stabilizers, solubilizers, vitamins, pigments and flavorings. And a carrier.
  • ingredients conventionally used in external preparations for skin and / or cosmetics such as conventional adjuvants such as antioxidants, stabilizers, solubilizers, vitamins, pigments and flavorings.
  • a carrier e.g., a carrier.
  • other ingredients may be appropriately selected and blended by those skilled in the art without difficulty according to the kind or purpose of use of the external preparation for skin and / or cosmetic composition.
  • Skin cosmetic method of an embodiment of the present invention (c) performing iontophoresis (iontophoresis) by flowing a microcurrent to the skin of the mammal to which the composition containing the stem cell-derived exosomes as an active ingredient is applied And (d) delivering the stem cell-derived exosomes into the mammalian skin through the microcurrent.
  • iontophoresis iontophoresis
  • the composition is, for example, patches, mask packs, mask sheets, creams, tonics, ointments, suspensions, emulsions, pastes, lotions, gels, oils, packs, sprays, It can be applied to various forms such as aerosol, mist, foundation, powder and oil paper.
  • the composition may be applied or deposited on at least one surface of the first mask pack, mask sheet or patch.
  • the step (b) is (b1) applying the composition directly to the skin of the mammal, (b2) the first mask pack to which the composition is applied or deposited, Contacting or attaching a masksheet or patch to the skin of the mammal or by sequentially proceeding with (b1) and (b2).
  • At least one surface of the first mask pack, mask sheet, or patch may include a hydrogel, hyaluronic acid, hyaluronic acid salt (for example, sodium hyaluronate), or hyaluronic acid.
  • hyaluronic acid for example, sodium hyaluronate
  • hyaluronic acid salt for example, sodium hyaluronate
  • hyaluronic acid for example, sodium hyaluronate
  • hyaluronic acid for example, sodium hyaluronate
  • hyaluronic acid for example, sodium hyaluronate
  • hyaluronic acid for example, sodium hyaluronate
  • hyaluronic acid for example, sodium hyaluronate
  • hyaluronic acid for example, sodium hyaluronate
  • hyaluronic acid for example, sodium hyaluronate
  • hyaluronic acid for example, sodium
  • step (c) may be performed by contacting or attaching an iontophoresis device to the skin of the mammal.
  • the iontophoresis device is a flexible battery, lithium ion secondary battery, alkaline battery, dry cell, mercury battery, lithium battery, nickel-cadmium battery, and reverse electrodialysis battery It may include at least one battery selected from the group consisting of, or may be a second mask pack, mask sheet or patch on which the at least one battery is mounted. Step (c) may be performed by stacking the second mask pack, mask sheet or patch on the first mask pack, mask sheet or patch.
  • Skin beauty method of the present invention by treating the skin with exosomes derived from stem cells with the laser beam irradiation to reduce the redness of the skin, which is a side effect of the laser beam irradiation, pain reduction and skin sedation due to the laser beam irradiation ( soothing) effect.
  • downtime which is a time taken to disappear the skin redness and swelling which is a side effect of the laser beam irradiation Not only that, but also a positive skin cosmetic effect such as skin pore reduction effect and facial radiance area increase.
  • the skin care method of the present invention improves the side effect reduction effect by laser beam irradiation and the positive skin care effect as described above when applying exosomes derived from stem cells to the skin using an iontophoresis device. You can.
  • FIG. 1 is a flowchart illustrating a process for separating and purifying exosomes in a method for producing exosomes from stem cell culture according to one embodiment of the present invention.
  • Figure 2 shows the results of measuring the relative amount of protein (Relative amount of protein) contained in the solution for each step (step) to prepare an exosome from the stem cell culture in accordance with an embodiment of the present invention.
  • the ratio of the total amount of protein in each step is expressed as the relative ratio of the total amount of protein to the stem cell culture.
  • the experimental results show the results obtained in two different batches, respectively.
  • Figure 3 shows the results of measuring the productivity (purity) and (productivity) of the exo-some obtained in accordance with an embodiment of the present invention.
  • the productivity of the exosomes was calculated as "the number of particles of exosomes per mL of stem cell culture (CM)", and the purity of the exosomes was calculated as "the number of particles of exosomes per ⁇ g of protein contained in the final fraction”. It was.
  • the experimental results show the results obtained in five different batches.
  • 4A to 4E show the results of physical characterization of the exosomes obtained according to one embodiment of the present invention.
  • 4A shows particle size distribution and particle number by tunable resistive pulse sensing (TRPS) analysis.
  • 4B shows particle size distribution and particle number by NTA (nanoparticle tracking analysis) analysis.
  • FIG. 4C shows the particle image by magnification by means of the transmitted electron microscopy (TEM) analysis.
  • TEM transmitted electron microscopy
  • 4D shows Western blot results of exosomes obtained according to one embodiment of the invention.
  • 4E shows the results of flow cytometry for CD63 and CD81 in marker analysis for exosomes obtained according to one embodiment of the invention.
  • 5A-5C show NTA analysis results for particle size distribution showing that exosomes with uniform particle size distribution and high purity are obtained with trehalose addition. As the amount of trehalose added increases, particle size distribution results with a single peak can be obtained.
  • FIGS. 6A to 6C show NTA analysis results showing particle size distribution depending on whether trehalose is added in the preparation of exosomes according to one embodiment of the present invention.
  • FIG. 6A shows the addition of trehalose throughout the manufacturing process
  • FIG. 6B shows freezing of the cell culture and thawing after thawing
  • FIG. 6C shows trehalo. The result obtained without adding oss is shown.
  • 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 particle size of the exosomes isolated by the methods of FIGS. 6A-6C.
  • Figure 7 shows the results of fluorescence intensity measurements confirming exosomes stained with PKH67.
  • Figure 8 is a fluorescence microscope image confirming the degree of delivery of the fluorescent stained exosomes inside the pig skin tissue.
  • 9 is a confocal fluorescence microscope image confirming the degree of delivery of fluorescently stained exosomes into mouse skin tissue.
  • FIG. 10 is a graph comparing total fluorescence intensities obtained by measuring fluorescence intensities on each image of FIG. 9.
  • FIG. 11 is a photograph comparing a skin condition of a human body treated with a composition comprising a stem cell-derived exosome according to an embodiment of the present invention after laser beam irradiation and before and after treatment.
  • FIG. 11 is a photograph comparing a skin condition of a human body treated with a composition comprising a stem cell-derived exosome according to an embodiment of the present invention after laser beam irradiation and before and after treatment.
  • FIG. 12A shows the right side of the face coated with a stem cell-derived exosome according to an embodiment of the present invention after laser beam irradiation, and the left side of the face coated with physiological saline after laser beam irradiation.
  • Figure 12B shows the result of measuring the redness after applying a exosome and physiological saline stem-derived physiological saline in accordance with an embodiment of the present invention to the right and left of the face after a laser beam irradiation for a certain time, respectively The figure is shown by zone.
  • FIG. 13 is a red phase on day 1 and day 3 after applying exosomes and physiological saline derived from stem cells according to an embodiment of the present invention to the right and left sides of the subject 1 after Er: Yag laser beam irradiation. redness) is a graph of results.
  • FIG. 14 shows redness at day 14 after applying exosomes and physiological saline derived from stem cells according to one embodiment of the present invention to the right and left sides of Subject 2 after Er: Yag laser beam irradiation. It is a graph of the measured result.
  • FIG. 17 after the Pico laser beam was applied to the stem cell-derived exosomes and vitamin C according to an embodiment of the present invention to the right and left of the subject 4 respectively, 18 days after further performing iontophoresis
  • the graph shows the result of measuring skin pore size.
  • the graph shows the result of measuring the radiance of the face.
  • Figure 19 shows the results confirmed that there is no cytotoxic after treating the stem cell-derived exosomes according to an embodiment of the present invention to HS68 cells, which are human skin fibroblasts.
  • 20 is a perspective view showing the configuration of an exosome kit 100 according to an embodiment of the present invention.
  • HS68 cells which are human dermal fibroblasts, are purchased from ATCC and are prepared by 10% fetal bovine serum (purchased from ThermoFisher Scientific) and 1% antibiotic-antimycotics (purchased from ThermoFisher Scientific). Passage was carried out in DMEM (purchased from ThermoFisher Scientific) medium containing 5% CO 2 at 37 ° C.
  • Fat-derived stem cells were cultured at 5% CO 2 , 37 ° C. according to cell culture methods known in the art. Then, washed with phosphate-buffered saline (purchased from ThermoFisher Scientific), replaced with serum-free, phenol-free medium, cultured for 1 to 10 days, and the supernatant (hereinafter, culture) was recovered. .
  • phosphate-buffered saline purchased from ThermoFisher Scientific
  • exosomes In the separation of exosomes, 2% by weight of trehalose was added to the culture to obtain exosomes with uniform particle size distribution and high purity. After the addition of trehalose, the culture solution was filtered through a 0.22 ⁇ m filter to remove impurities such as cell debris, waste, and large particles. The filtered culture immediately separated the exosomes through a separation process. In addition, the filtered culture was stored in the refrigerator (image 10 °C or less) and then used for exosome separation. In addition, the filtered culture solution was stored frozen in an cryogenic freezer of -60 °C or less and thawed and then exosomes were separated. Thereafter, exosomes were separated from the culture using a tangential flow filtration device (TFF).
  • TMF tangential flow filtration device
  • Example 1 the exosomes were separated from the culture medium filtered with a 0.22 ⁇ m filter, and the TFF (Tangential Flow Filtration) method was used for concentration, desalting and diafiltration.
  • the filter for the TFF method was a cartridge filter (aka hollow fiber filter; purchased from GE Healthcare) or a cassette filter (purchased from Pall or Sartorius or Merck Millipore).
  • TFF filters can be selected by various molecular weight cutoffs (MWCO). Exosomes were selectively isolated and concentrated by the selected MWCO, and particles, proteins, lipids, nucleic acids, and small molecule compounds smaller than MWCO were removed.
  • MWCO molecular weight cutoffs
  • MWCO 100,000 Da (Dalton), 300,000 Da, or 500,000 Da TFF filter was used to isolate and concentrate the exosomes.
  • the culture solution was concentrated to a volume of 1/100 to 1/25 by using the TFF method, while exosomes were separated by removing substances smaller than MWCO.
  • the separated and concentrated exosome solution was further subjected to desalting and diafiltration using the TFF method.
  • desalting and buffer exchange were carried out continuously (discontinuous diafiltration) or at least 4 times, preferably 6 times to 10 times, more preferably, relative to the starting volume. It was performed using a buffer solution having a volume of 12 times or more.
  • To the buffer solution was added 2% by weight of trehalose dissolved in PBS to obtain exosomes with uniform particle size distribution and high purity.
  • Figures 6A to 6E The results of confirming the effect of obtaining a high purity and uniform particle size distribution of exosomes according to the trehalose treatment in a high yield are shown in Figures 6A to 6E.
  • the amount of protein in the isolated exosomes, cultures, and fractions of TFF separation was measured using BCA coloration (purchased from ThermoFisher Scientific) or FluoroProfile fluorescence (purchased from Sigma).
  • Exosome is isolated and concentrated by the TFF method of one embodiment of the present invention, and the degree of protein, lipid, nucleic acid, low molecular weight compounds, etc. is monitored by protein quantitation and the results are shown in FIG. As a result, it was found that the protein present in the culture medium was effectively removed by the TFF method of one embodiment of the present invention.
  • the isolated exosomes were measured for particle size and concentration by nanoparticle tracking analysis (NTA; purchased from Malvern) or tunable resistive pulse sensing (TRPS; purchased from Izon Science).
  • NTA nanoparticle tracking analysis
  • TRPS tunable resistive pulse sensing
  • the uniformity and size of the isolated exosomes were analyzed using a transmitted electron microscopy (TEM).
  • TRPS, NTA, TEM analysis results of the exosomes isolated in accordance with one embodiment of the present invention are shown in Figures 4A to 4C.
  • FIGS. 5A to 5C the results of NTA analysis of the size distribution of the exosomes depending on whether trehalose was added are shown in FIGS. 5A to 5C.
  • Trehalose concentrations were increased to 0%, 1% and 2% by weight (from top to bottom in FIGS. 5A-5C) and were repeated three times.
  • trehalose is not present, particles having a size of 300 nm or more are identified, while increasing the amount of trehalose added decreases the particles having a size of 300 nm or more and makes the size distribution of exosomes uniform. .
  • Figure 4D shows the results of Western blot for exosomes isolated according to the method of one embodiment of the present invention, confirming the presence of CD9, CD63, CD81 and TSG101 markers.
  • Anti-CD9 purchased from Abcam
  • anti-CD63 purchasedd from System Biosciences
  • anti-CD81 purchasedd from System Biosciences
  • anti-TSG101 purchasedd from Abcam
  • Figure 4E confirmed the presence of CD63 and CD81 markers as a result of analysis using a flow cytometer for the exosomes isolated in accordance with the method of one embodiment of the present invention.
  • an exosome-human CD63 separation / detection kit purchased from ThermoFisher Scientific
  • PE-mouse anti-human CD63 PE-Mouse anti markers were stained using -human CD63
  • PE-mouse anti-human CD81 purchasedd from BD
  • the present invention confirms that exosomes with high purity and uniform particle size distribution can be efficiently and efficiently separated and purified in high yield by adding trehalose in the manufacturing process using tangential flow filtration.
  • the processes of the separation method of one embodiment of the present invention are scale-up and suitable for GMP.
  • HS68 cells which are human skin fibroblasts
  • HS68 cells were treated with exosomes at different concentrations, and cell proliferation rates were confirmed.
  • HS68 cells were suspended in DMEM containing 10% FBS and then aliquoted to have a confluency of 80-90% and incubated in 37 ° C., 5% CO 2 incubator for 24 hours. After 24 hours, the culture solution was removed and the exosomes prepared in Example 2 were treated for each concentration, and cultured for 24 to 72 hours to evaluate cell viability.
  • WST-1 reagent purchased from Takara
  • MTT reagent purchased from Sigma
  • CellTiter-Glo reagent purchased from Promega
  • Aramamar blue reagent Measurements were performed using alamarBlue reagent (purchased from ThermoFisher Scientific) and 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 exosomes, it was confirmed that no cytotoxicity by the exosomes of the present invention within the tested concentration range (Fig. 19).
  • 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, and then mixed with an appropriate concentration of exosome solution and reacted for 10 minutes while blocking light at room temperature. After the reaction, a MW3000 spin column (purchased from ThermoFisher Scientific) was used to remove residual PKH67 dye from exosomes stained with PKH67 (hereinafter abbreviated as "PKH-exosome").
  • PKH-exosomes were dispersed in phosphate buffer solution (PBS) at a suitable concentration, for example 1 ⁇ 10 5 particles / mL to 1 ⁇ 10 9 particles / mL, and then applied to the outside of pig skin. To prevent drying of the applied PKH-exosomes, the nonwoven fabric was covered and then reacted for an appropriate time, for example, 30 minutes to 1 hour, so that the PKH-exosomes were delivered to the subcutaneous tissue. In addition, PKH-exosomes were applied to the outside of the pig skin and covered with a nonwoven fabric, and then a microcurrent was flowed for a predetermined time, for example, 30 minutes to 1 hour.
  • PBS phosphate buffer solution
  • the pig skin tissue was immersed in 3.7% formaldehyde solution to react overnight, washed three times with phosphate buffer solution for 5 minutes.
  • the washed porcine skin tissue was soaked in 30% sucrose solution and then treated with OCT compound.
  • the fabrication of tissue sections can also be performed prior to fixation of the tissue with formaldehyde solution. Fluorescence detected from PKH-exosomes in tissue sections was observed using a fluorescence microscope. The above method confirmed the PKH-exosomes that penetrated the epidermis of the pig skin tissue and delivered to the subcutaneous tissue.
  • the exosomes more effectively passed through the epidermis and deeply delivered into the skin tissue, it was confirmed that more effectively absorbed by the skin (Fig. 8).
  • the skin tissue of the hairless mouse was extracted and placed above the chamber of the Franz Diffusion Cell.
  • the inside of the diffusion cell was filled with a phosphate buffer solution.
  • PKH-exosomes were dispersed in phosphate buffer solution (PBS) at a suitable concentration, for example 1 ⁇ 10 5 particles / mL to 1 ⁇ 10 9 particles / mL, and then applied to the outside of the skin tissue of the mouse.
  • PBS phosphate buffer solution
  • the nonwoven fabric was pre-positioned outside the mouse skin tissue, and PKH-exosomes were injected between the nonwoven fabric and the skin tissue. After that, the reaction was carried out for 30 minutes to 1 hour.
  • a microcurrent was flowed for a predetermined time, for example, 30 minutes to 1 hour. After the reaction, immediately check the PKH-exosomes delivered into the skin tissue using confocal fluorescence microscope (Leica, SP8X), or further react the skin tissue and PKH-exosome solution for 1 to 6 hours. After PKH-exosomes were confirmed by confocal fluorescence microscopy. As a result, in the case of the micro current flowing (active transdermal delivery), it was confirmed that the exosomes more effectively passed through the epidermis and deeply into the skin tissue, and more effectively absorbed by the skin (FIGS. 9 and 10). .
  • the experimental results show that when exosomes derived from stem cells are applied to the skin while flowing microcurrent through the skin by iontophoresis, the exosomes derived from stem cells effectively pass through the epidermis and deep into the skin tissue. Can be delivered and effectively absorbed by the skin. Therefore, when the exosomes derived from stem cells are applied to the skin using an iontophoresis device (an apparatus for flowing a microcurrent to the skin) and / or the exosome kit 100 including the same, the skin penetration ability is higher. It can be improved to enhance the side effect reduction effect by the laser beam irradiation and the positive skin cosmetic effect as described above.
  • the exosome derived from the stem cell of one embodiment of the present invention may be directly applied to the skin, and an iontophoresis device (active transdermal permeation device for flowing a microcurrent to the skin) or It may be applied to the skin using the exosome kit 100 having it.
  • the photograph immediately before the laser beam irradiation (Before) and the photograph (After) three days after the application of the stem cell-derived exosome of one embodiment of the present invention were compared (see FIG. 11).
  • Erbium Fraxel Laser was irradiated on the human face using an Er: YAG laser instrument, Joule (Sciton, Inc., Calif., USA).
  • the laser parameters were as follows: wavelength 2940 nm; Wavelength energy 7 J / cm 2 ; 10 ⁇ 10 cm scanning mode; 1 pass].
  • anesthesia was applied to the face for 30 minutes and irradiated with an erb Fraxel laser.
  • 1 ml of an exosome (exosome prepared in Example 2) derived from stem cells at a concentration of 7.39 ⁇ 10 8 particles / mL was placed on the right half of each subject's face immediately after laser beam irradiation (suspension).
  • uspension was applied, and an appropriate amount of physiological saline was applied to the left half of each subject's face.
  • a rubber mask was laminated and pressed on the face of each subject for 30 minutes.
  • the redness of each subject's face was measured using a Mark-Vu facial skin analyzer of PS-I Plus, Gyeonggi-do, Korea. Before the laser beam irradiation (Before), and after applying the stem cell-derived exosomes of one embodiment of the present invention for measuring the red phase (After) a day and 3 days after (After) Compared. Based on the results of the red phase measurement, when the exosome derived from the stem cell of the present invention is treated with the laser beam irradiation on the skin, it was evaluated whether the side effects due to the laser beam irradiation were alleviated or restored to the normal state.
  • the stem cell-derived exosomes and physiological saline according to one embodiment of the present invention were applied to the right and left sides of the subject 1, respectively, on the first and third days.
  • the exosomes and the physiological saline derived from the stem cells of one embodiment of the present invention were respectively applied to the right and left sides of the subject 2, and then the mark-view facial skin Redness was measured using an analyzer.
  • the redness of the E region and the G region of the face region treated with the exosomes derived from the stem cells of one embodiment of the present invention was significantly reduced compared to the F and H regions of the face region to which the physiological saline was applied, respectively. It could be confirmed that (Fig. 14).
  • the stem cell-derived exosomes (7.39 ⁇ 10 8 particles / mL concentration) of one embodiment of the present invention were applied to the right side of the subject 3's face. . Iontophoresis was performed for 15 minutes on the right face, and a rubber mask was laminated and pressed on the left face for 15 minutes. Iontoporesis was performed by flowing a 0.5 mA microcurrent for 20 minutes to the right face of the stem cell-derived exosomes using an iontophoresis device (IONZYME) (purchased from Environ). The red phases of the right and left faces before and after the treatment were measured and compared in the manner described in Example 7-1. As a result, when applying exosomes derived from stem cells to the face and performing iontophoresis, it was confirmed that the redness immediately decreased significantly compared to simply pressing and stacking a rubber mask (FIG. 15).
  • IONZYME iontophoresis device
  • the exosomes 12 derived from the stem cells were applied using the IONZYME iontophoresis device, but immediately after the laser beam irradiation, the exosomes 12 derived from the stem cells
  • the liquid immersion sheet mask 10 included in the liquid immersion sheet mask 10 is adhered evenly to the entire face of the person in accordance with the eyes and mouth, and then the liquid transdermal penetration promotion sheet mask 20 is activated to flow a micro current of about 0.3 to 0.4 mA.
  • the iontophoresis may be performed by laminating on the sheet mask 10 and using it for about 25 to 30 minutes (FIG. 20).
  • the Pico laser was irradiated on the face of a person using Pico Plus, Pico Plus, Lutronic, Inc., Gyeonggi-do, Korea.
  • Laser parameters are as follows: wavelength 1064 nm; Wavelength energy 0.7 J / cm 2 ; Laser spot size 7 mm; Laser irradiation time 450 ps (picosecond); Frequency 10 Hz; Number of surveys 3000 shots].
  • exosomes prepared in Example 2 were applied to the right face of Subject 4 with a concentration of 7.39 ⁇ 10 8 particles / mL of the stock solution (suspension), and left of Subject 4 A proper amount of vitamin C solution (1% vitamin phosphate solution) was applied to the face. Then, iontophoresis was performed on the right and left faces of the subject 4 in the same manner as described in Example 7-2.

Abstract

La présente invention concerne un procédé pour soins de la peau comprenant : (a) une étape d'exposition de la peau d'un mammifère à un faisceau laser ; et (b) une étape d'application, avant ou après l'étape d'exposition au faisceau laser, d'une composition contenant un exosome dérivé de cellules souches à titre de principe actif sur la peau du mammifère ayant été soumise à l'exposition au faisceau laser. Le traitement par exosomes dérivés de cellules souches conjointement avec l'exposition au faisceau laser, selon le procédé pour soins de la peau de la présente invention, permet de raccourcir le temps d'arrêt, qui est le temps nécessaire pour que les phénomènes de rougeur et de gonflement de la peau, qui sont des effets secondaires provoqués par l'exposition au faisceau laser, disparaissent, et permet également d'exercer les effets de soins de la peau bénéfiques d'une réduction des pores cutanés et d'un accroissement d'une zone de radiance faciale.
PCT/KR2019/001677 2018-03-02 2019-02-12 Procédé pour soins de la peau par exposition de la peau à un faisceau laser et traitement par exosomes dérivés de cellules souches WO2019168281A1 (fr)

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
KR10-2018-0025100 2018-03-02
KR20180025100 2018-03-02
KR10-2018-0062707 2018-05-31
KR20180062707 2018-05-31

Publications (1)

Publication Number Publication Date
WO2019168281A1 true WO2019168281A1 (fr) 2019-09-06

Family

ID=67806295

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/KR2019/001677 WO2019168281A1 (fr) 2018-03-02 2019-02-12 Procédé pour soins de la peau par exposition de la peau à un faisceau laser et traitement par exosomes dérivés de cellules souches

Country Status (2)

Country Link
KR (1) KR20190104881A (fr)
WO (1) WO2019168281A1 (fr)

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
USD891628S1 (en) 2015-03-03 2020-07-28 Carol Cole Company Skin toning device
CN112553152A (zh) * 2020-10-27 2021-03-26 重庆市铂而斐细胞生物技术有限公司 一种快速提高脂肪间充质干细胞外泌体产量的方法
USD949358S1 (en) 2018-05-15 2022-04-19 Carol Cole Company Elongated skin toning device
USD953553S1 (en) 2020-02-19 2022-05-31 Carol Cole Company Skin toning device
USD957664S1 (en) 2020-07-29 2022-07-12 Carol Cole Company Skin toning device

Families Citing this family (1)

* 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

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20100122087A (ko) * 2008-02-22 2010-11-19 에이전시 포 사이언스, 테크놀로지 앤드 리서치 중간엽 줄기세포 입자
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
KR20170082111A (ko) * 2016-01-05 2017-07-13 바이오센서연구소 주식회사 마스크팩에 장착되는 장치, 그를 포함하는 마스크팩 및 키트
JP2017520555A (ja) * 2014-07-03 2017-07-27 ピープラス スキン ケア リミテッド 美容方法および美容製品

Family Cites Families (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR101015881B1 (ko) 2010-06-17 2011-02-23 이용수 레이저를 이용한 흉터 및 피부질환 치료 시스템 및 그 방법
KR101761680B1 (ko) 2015-03-31 2017-08-23 포항공과대학교 산학협력단 수용액 이상계를 이용한 세포 밖 소포체의 분리방법

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20100122087A (ko) * 2008-02-22 2010-11-19 에이전시 포 사이언스, 테크놀로지 앤드 리서치 중간엽 줄기세포 입자
JP2017520555A (ja) * 2014-07-03 2017-07-27 ピープラス スキン ケア リミテッド 美容方法および美容製品
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
KR20170082111A (ko) * 2016-01-05 2017-07-13 바이오센서연구소 주식회사 마스크팩에 장착되는 장치, 그를 포함하는 마스크팩 및 키트

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
22 October 2015 (2015-10-22), Retrieved from the Internet <URL:https://blog.naver.com/restemkorea/220516198077> *

Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
USD891628S1 (en) 2015-03-03 2020-07-28 Carol Cole Company Skin toning device
USD949358S1 (en) 2018-05-15 2022-04-19 Carol Cole Company Elongated skin toning device
USD959005S1 (en) 2018-05-15 2022-07-26 Carol Cole Company Elongated skin toning device
USD953553S1 (en) 2020-02-19 2022-05-31 Carol Cole Company Skin toning device
USD957664S1 (en) 2020-07-29 2022-07-12 Carol Cole Company Skin toning device
USD1017822S1 (en) 2020-07-29 2024-03-12 Carol Cole Company Skin toning device
CN112553152A (zh) * 2020-10-27 2021-03-26 重庆市铂而斐细胞生物技术有限公司 一种快速提高脂肪间充质干细胞外泌体产量的方法

Also Published As

Publication number Publication date
KR20190104881A (ko) 2019-09-11

Similar Documents

Publication Publication Date Title
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
KR102359387B1 (ko) 줄기세포 유래의 엑소좀을 유효성분으로 포함하는 피지분비 감소용 조성물
WO2019182299A1 (fr) Procédé de soin de la peau utilisant le rayonnement ipl sur la peau et traitement d&#39;exosome dérivé de cellules souches en combinaison
KR102646145B1 (ko) 줄기세포 유래의 엑소좀을 유효성분으로 포함하는 모공 축소용 조성물
EP3639832A2 (fr) Utilisation d&#39;une composition comprenant un exosome dérivé de cellules souches adipeuses en tant que principe actif pour atténuer la dermatite
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
KR102045188B1 (ko) 지방줄기세포 유래의 엑소좀을 유효성분으로 포함하는 조성물의 피부염 개선 용도
KR20190069301A (ko) 줄기세포 유래의 엑소좀을 유효성분으로 포함하는 민감성 피부의 예방, 억제, 완화 또는 개선용 화장료 조성물
KR20190003399A (ko) 줄기세포 유래의 엑소좀을 유효성분으로 포함하는 조성물의 피부 섬유증 개선 용도
WO2021033899A1 (fr) Composition pour la cicatrisation ou l&#39;accélération de la cicatrisation, contenant, en tant que principe actif, une combinaison d&#39;acide hyaluronique et d&#39;exosomes lyophilisés dérivés de cellules souches
KR20190069277A (ko) 줄기세포 유래의 엑소좀 및/또는 세포외 소포체를 유효성분으로 포함하는 조성물의 민감성 피부 예방, 억제, 완화 또는 개선 용도
KR20240037208A (ko) 줄기세포 유래의 엑소좀을 유효성분으로 포함하는 안면 홍조 개선용 조성물
WO2019088545A1 (fr) Kit d&#39;exosomes et procédé d&#39;amélioration de la perméation transdermique d&#39;exosomes l&#39;utilisant
WO2021201637A1 (fr) Procédé de préparation d&#39;un milieu de culture contenant des niveaux élevés d&#39;exosomes hautement puissants sécrétés par des cellules souches de sang de cordon ombilical et utilisation correspondante
KR20190116858A (ko) 줄기세포 유래의 엑소좀 및/또는 세포외 소포체를 유효성분으로 포함하는 조성물의 염증후 과색소침착의 예방, 완화, 감소 내지는 제거 용도
WO2019112238A2 (fr) Composition cosmétique pour hydrater la peau comprenant un exosome dérivé de cellules souches en tant qu&#39;ingrédient actif
WO2020027467A1 (fr) Préparation lyophilisée d&#39;exosomes issus de cellules souches et composition anti-inflammatoire la comprenant en tant que principe actif
WO2019190093A1 (fr) Procédé de soin de la peau utilisant, en combinaison, l&#39;application d&#39;énergie rf sur la peau et le traitement par exosomes dérivés de cellules souches sur la peau
KR20130021793A (ko) 지방 유래 성체줄기세포 배양액의 나노섬유 또는 나노캡슐을 포함하는 화장료 조성물 및 그 제조방법
KR20200016183A (ko) 피부 필링과 줄기세포 유래의 엑소좀 처리를 병용한 피부 미용방법
KR20200038672A (ko) 피부에 대한 플라즈마 처리와 엑소좀 처리를 병용한 피부 상태의 개선 방법
WO2019231133A1 (fr) Composition pour réduire les pores, comprenant des exosomes dérivés de cellules souches en tant que principe actif
KR20190114740A (ko) 피부에 대한 ｒｆ 에너지 인가와 줄기세포 유래의 엑소좀 처리를 병용한 피부 미용방법
KR20190045829A (ko) 엑소좀의 경피 투과 증진 방법 및 이의 응용
WO2019112229A2 (fr) Nouvelle utilisation d&#39;un kit d&#39;exosomes comprenant un exosome dérivé d&#39;une cellule souche

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: 19761630

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 19761630

Country of ref document: EP

Kind code of ref document: A1