Composition containing stem cell exosome as well as preparation method and application thereof in cosmetics

Abstract

The invention discloses a composition containing stem cell exosomes, a preparation method of the composition and application of the composition in cosmetics, and belongs to the technical field of biology. Culturing the mesenchymal stem cells, collecting supernate, and extracting exosomes; preparing a chamomile extract, a white willow bark extract and a centella asiatica extract; the exosome, the chamomile extract, the white willow bark extract, the centella asiatica extract, PCA zinc, salicylic acid, panthenol and sodium hyaluronate are stirred and mixed uniformly, and the composition containing the stem cell extracting solution is prepared. The composition containing the stem cell exosome prepared by the invention can form a synergistic interaction system of the exosome and various components, realizes anti-inflammatory repair synergy, cortical regulation synergy and micro-ecological synergy through a multi-target action mechanism, has an outstanding effect on inhibition of propionibacterium acnes and inhibition of skin grease, and has a wide application prospect.

Description

Technical Field

[0001] The present invention relates to the field of biotechnology, and in particular to a composition containing stem cell exosomes, a preparation method thereof, and an application in cosmetics. Background Art

[0002] With the development of social economy and the upgrading of consumption concepts, the global cosmetics market scale has been continuously expanding. In 2023, the global anti-acne product market has reached 18.7 billion US dollars, with a compound annual growth rate of 5.8%, reflecting that acne has become a common skin problem affecting 450 million global patients. It is worth noting that the acne incidence group shows a significant trend of adultization, which is closely related to modern life stress, environmental pollution, and changes in dietary structure. Affected by the aggravation of environmental pollution, the popularization of high-sugar diets, and the doubling of workplace stress, sebum secretion imbalance has become the core cause of modern skin problems. Clinical data shows that about 42% of adults globally suffer from persistent oily skin problems. Among them, due to the high temperature and high humidity climate characteristics in the Asian region, the sebum secretion in the T-zone is 1.3 - 1.8 times higher than that of the European and American populations.

[0003] There are bottlenecks in existing solutions: Firstly, although chemically synthesized ingredients can quickly inhibit sebum secretion, 28% of users experience impaired barrier function; Secondly, single-effect ingredients are difficult to simultaneously solve multi-dimensional pathogenic mechanisms such as sebum hypersecretion, Propionibacterium acnes proliferation, and abnormal follicular keratinization. Consumer research reveals new demand trends: 62% of acne patients prefer "oil control - acne removal - repair" trinity products, and 79% tend to formulas containing natural active ingredients. Summary of the Invention

[0004] The purpose of the present invention is to provide a composition containing stem cell exosomes, a preparation method thereof, and an application in cosmetics, so as to provide a product with outstanding effects in inhibiting Propionibacterium acnes and skin oil.

[0005] The technical solution of the present invention is realized as follows: In the first aspect, the present invention provides a preparation method of a composition containing stem cell exosomes, including the following steps: Step S1: Culture of mesenchymal stem cells: Culture mesenchymal stem cells with MSCGM-CD™ medium. When the cell confluence reaches 80% - 90%, discard the medium, wash with PBS buffer, add 2 mL of trypsin-EDTA for digestion. After terminating the digestion, gently pipette to form a single-cell suspension, centrifuge and discard the supernatant, then resuspend with fresh MSCGM-CD™ medium, inoculate into a new culture flask. When the cell confluence reaches 60% - 70%, centrifuge and collect the supernatant; Step S2: Extraction of exosomes: Transfer the supernatant obtained in Step S1 to a new tube, centrifuge to remove dead cells and debris, filter the supernatant to remove residual particles, dispense the filtrate into an ultracentrifuge tube, centrifuge and discard the supernatant, retain the precipitate at the bottom of the tube, resuspend the precipitate with pre-cooled PBS buffer, gently pipette, centrifuge and discard the supernatant, and resuspend the precipitate with sterile PBS buffer to obtain exosomes; Step S3: Preparation of chamomile, white willow bark and centella asiatica extracts: Take chamomile, crush and sieve it, mix the obtained chamomile powder with 70% ethanol, impregnate at room temperature for 30 minutes, perform ultrasonic extraction, after the ultrasonic extraction ends, centrifuge, collect the supernatant, concentrate the supernatant with a rotary evaporator, and freeze-dry it into a powder to obtain chamomile extract; Obtain white willow bark extract and centella asiatica extract respectively according to the same operation as chamomile; Step S4: Preparation of the composition containing stem cell exosomes: Stir and mix zinc PCA, salicylic acid, panthenol, sodium hyaluronate, the exosomes described in Step S2, the chamomile extract, white willow bark extract and centella asiatica extract described in Step S3 for 30 minutes to prepare a composition containing stem cell exosomes.

[0006] As a further improvement of the present invention, the mesenchymal stem cells described in Step S1 are human bone marrow mesenchymal stem cells.

[0007] As a further improvement of the present invention, the concentration of trypsin-EDTA described in Step S1 is 2%-3%.

[0008] As a further improvement of the present invention, the mass of zinc PCA described in Step S4 is 1-2 g, the mass of salicylic acid is 3-6 g, the mass of panthenol is 4-8 g, the mass of sodium hyaluronate is 1-2 g, the mass of chamomile extract is 1-2 g, the mass of white willow bark extract is 1-2 g, the mass of centella asiatica extract is 1-2 g, and the mass of exosomes is 2-4 g.

[0009] The present invention further protects a composition containing stem cell exosomes prepared by the above preparation method.

[0010] The present invention further also protects the application of the above composition containing stem cell exosomes in cosmetics.

[0011] The present invention has the following beneficial effects: The present invention adds exosomes, chamomile extract, white willow bark extract, centella asiatica extract and zinc PCA to the composition. Exosomes are 30-150 nm bilayer lipid vesicles secreted by mesenchymal stem cells. As the core carrier of intercellular communication, they carry a large number of active ingredients, such as heat shock proteins and transmembrane proteins, and participate in cell stress protection and targeted recognition. At the same time, exosomes carry miRNA-21-5p, miRNA-29a-3p, etc., which inhibit the expression of inflammatory factors through epigenetic regulation. In addition, prostaglandin E2 and sphingosine-1-phosphate contained in exosomes can directly regulate the expression of the COL1A1 gene in skin fibroblasts. Chamomile extract has a soothing effect and can repair sensitive skin. White willow bark extract can dredge pores, help remove excess fat, stains and skin cutin, and reduce skin inflammation. Centella asiatica extract has a bactericidal and anti-inflammatory effect and can promote skin cell regeneration. Zinc PCA can regulate the secretion of sebaceous glands, inhibit the activity of 5α-reductase, and reduce oil synthesis.

[0012] The present invention innovatively constructs a synergistic system of exosomes and multiple components, and improves the effects of the composition in inhibiting Propionibacterium acnes and skin oil through a multi-target action mechanism. The specific synergistic effects are reflected in the following aspects: (1) Anti-inflammatory and repair synergistic pathway: miRNA-21-5p carried by exosomes down-regulates pro-inflammatory factors such as IL-6 and TNF-α by inhibiting the TLR4 / NF-κB pathway, and the bisabolol in chamomile extract inhibits the release of histamine from mast cells to form a dual anti-inflammatory mechanism. At the same time, prostaglandin E2 in exosomes activates the Wnt / β-catenin signal to promote wound repair, and asiaticoside enhances fibroblast migration through the TGF-β / Smad pathway to form a repair network; (2) Sebum regulation synergistic system: Zinc PCA reduces DHT-induced sebum secretion by inhibiting 5α-reductase, and sphingosine-1-phosphate in exosomes regulates the PPARγ pathway to reduce lipid synthesis to form a complement. Salicin in white willow bark extract enhances the permeability of zinc PCA while dissolving the keratin plug, making zinc ions act more effectively on sebaceous gland basal cells; (3) Microecological synergistic balance: Triterpenoid components in centella asiatica enhance the bactericidal effect by destroying the biofilm of Propionibacterium acnes, while the heat shock protein HSP70 in exosomes maintains the integrity of the skin barrier, and the two work together to reconstruct the skin microenvironment. In addition, exosomes, chamomile extract, white willow bark extract, centella asiatica extract, zinc PCA, sodium hyaluronate, salicylic acid and panthenol have good synergistic effects in maintaining keratin metabolism, barrier repair and redox metabolism. Therefore, the composition containing stem cell exosomes prepared by the present invention can play an outstanding role in inhibiting Propionibacterium acnes and skin oil, and has broad application prospects. Detailed implementation mode

[0013] The present invention will be further described in detail below in conjunction with specific embodiments. The provided embodiments are only for clarifying the present invention and not for limiting the scope of the present invention. The following provided embodiments can be used as a guide for those of ordinary skill in the art to make further improvements, and in no way constitute a limitation to the present invention.

[0014] Unless otherwise specified, the following embodiments do not include other components except inevitable impurities. The reagents and instruments used in the embodiments are all conventional selections in the art unless otherwise specified. For the experimental methods without specific conditions noted in the embodiments, they are carried out under conventional conditions, such as the conditions described in the literature, books, or the methods recommended by the manufacturer.

[0015] Example 1 Preparation of a Composition Containing Stem Cell Exosomes This example provides a method for preparing a composition containing stem cell exosomes, which includes the following steps: Step S1: Cultivation of bone marrow mesenchymal stem cells Disinfect the posterior superior iliac spine at the puncture site, and after local anesthesia, draw 10 mL of bone marrow fluid with a bone marrow puncture needle. Inject the bone marrow fluid into a centrifuge tube containing an anticoagulant, gently mix, dilute the bone marrow fluid with PBS buffer at a ratio of 1:1, filter it through a 100 μm cell sieve into a 50 mL centrifuge tube. Add 3 volumes of red blood cell lysate, let it stand at room temperature for 5 minutes, centrifuge at 300×g for 5 minutes, and discard the supernatant. Resuspend the cells with PBS buffer, centrifuge at 300×g for 5 minutes, and repeat 2 times. Resuspend the cells with 10 mL of MSCGM-CD™ medium, take 10 μL of the suspension for cell counting, and inoculate it into a T75 culture flask at a density of 5×10 5 cells / cm 2 and place it in an incubator. After culturing for 48 hours, discard the medium, gently add 5 mL of PBS buffer and wash 2 times to remove non-adherent cells, and then add 10 mL of MSCGM-CD™ medium. When the cell confluence reaches 80%, discard the medium and wash it once with PBS buffer. Add 2 mL of 0.2% trypsin-EDTA, digest at 37°C for 5 minutes, add MSCGM-CD™ medium to terminate digestion, gently pipette to obtain a single cell suspension, centrifuge at 300×g for 5 minutes, and discard the supernatant. Resuspend with MSCGM-CD™ medium, inoculate it into a new culture flask at a ratio of 1:3, and when the cell confluence reaches 60%, centrifuge at 300×g for 10 minutes and collect the supernatant. Step S2: Extraction of exosomes Transfer the supernatant to a new tube and centrifuge at 2000×g for 20 minutes to remove dead cells and debris. Filter the supernatant through a 0.22 μm filter membrane to remove residual particles. Aliquot the filtrate into ultracentrifuge tubes and centrifuge at 100000×g for 70 minutes at 4°C. Discard the supernatant and retain the pellet at the bottom of the tube. Resuspend the pellet with pre-chilled PBS buffer and gently pipette to avoid disrupting exosomes. Centrifuge at 100000×g for 70 minutes at 4°C and discard the supernatant. Resuspend the pellet with 200 μL of sterile PBS buffer to obtain exosomes derived from human bone marrow mesenchymal stem cells.

[0016] Step S3: Preparation of chamomile, white willow bark, and centella asiatica extracts Take 100 g of dried chamomile, crush it and sieve it through a 40-mesh sieve, and weigh it for later use. Mix the chamomile powder with 70% ethanol at a ratio of 1:15 (w / v), and soak it at room temperature for 30 minutes. Perform ultrasonic extraction with an ultrasonic power of 300 W, a temperature of 50°C, and an extraction time of 30 minutes. After ultrasonic extraction, centrifuge at 4000×g for 15 minutes and collect the supernatant. Concentrate the supernatant to 1 / 5 of the original volume using a rotary evaporator under the conditions of 50°C and a vacuum of -0.08 MPa. Lyophilize it into a powder to obtain chamomile extract.

[0017] According to the same method as above, weigh 100 g of white willow bark to prepare white willow bark extract; weigh 100 g of centella asiatica to prepare centella asiatica extract.

[0018] Step S4: Preparation of the composition containing stem cell exosomes Mix 1 g of chamomile extract, 1 g of white willow bark extract, 1 g of centella asiatica extract, 1 g of zinc PCA, 3 g of salicylic acid, 4 g of panthenol, 1 g of sodium hyaluronate, and 2 g of exosomes by stirring for 30 minutes to prepare the composition containing stem cell exosomes.

[0019] Example 2: Preparation of the composition containing stem cell exosomes This example provides a method for preparing a composition containing stem cell exosomes, including the following steps: Step S1: Culture of bone marrow mesenchymal stem cells Disinfect the posterior superior iliac spine at the puncture site, and after local anesthesia, aspirate 10 mL of bone marrow fluid with a bone marrow puncture needle. Inject the bone marrow fluid into a centrifuge tube containing anticoagulant, gently mix it, dilute the bone marrow fluid with PBS buffer at a ratio of 1:1, and filter it through a 100 μm cell sieve into a 50 mL centrifuge tube. Add 3 volumes of red blood cell lysate, let it stand at room temperature for 5 minutes, centrifuge at 300×g for 5 minutes, and discard the supernatant. Resuspend the cells with PBS buffer, centrifuge at 300×g for 5 minutes, and repeat 2 times. Resuspend the cells with 10 mL of MSCGM-CD™ medium, take 10 μL of the suspension for cell counting, and according to 5×105 cells / cm 2 Inoculate into a T75 culture flask and place it in an incubator. After culturing for 48 hours, discard the culture medium, gently add 5 mL of PBS buffer and wash twice to remove non-adherent cells, then add 10 mL of MSCGM-CD™ medium. When the cell confluence reaches 90%, discard the culture medium and wash once with PBS buffer. Add 2 mL of 0.3% trypsin-EDTA, digest at 37°C for 5 minutes, add MSCGM-CD™ medium to terminate digestion, gently pipette to form a single-cell suspension, centrifuge at 300×g for 5 minutes, and discard the supernatant. Resuspend with MSCGM-CD™ medium and inoculate into a new culture flask at a ratio of 1:3. When the cell confluence reaches 70%, centrifuge at 300×g for 10 minutes and collect the supernatant. Step S2: Extraction of exosomes Transfer the supernatant to a new tube, centrifuge at 2000×g for 20 minutes to remove dead cells and debris, filter the supernatant through a 0.22 μm filter membrane to remove residual particles. Aliquot the filtrate into an ultracentrifuge tube, centrifuge at 100000×g for 70 minutes at 4°C, discard the supernatant, and retain the precipitate at the bottom of the tube. Resuspend the precipitate with pre-cooled PBS buffer, gently pipette, avoiding damage to exosomes, centrifuge at 100000×g for 70 minutes at 4°C, and discard the supernatant. Resuspend the precipitate with 200 μL of sterile PBS buffer to obtain exosomes derived from human bone marrow mesenchymal stem cells.

[0020] Step S3: Preparation of chamomile, white willow bark, and centella asiatica extracts Take 100 g of dried chamomile, pulverize it and sieve through a 40-mesh sieve, weigh and set aside. Mix the chamomile powder with 70% ethanol at a ratio of 1:15 (w / v), impregnate at room temperature for 30 minutes. Perform ultrasonic extraction, set the ultrasonic power at 300 W, the temperature at 50°C, and the extraction time at 30 minutes. After ultrasonic extraction, centrifuge at 4000×g for 15 minutes and collect the supernatant. Concentrate the supernatant to 1 / 5 of the original volume using a rotary evaporator under the conditions of 50°C and a vacuum of -0.08 MPa. Lyophilize to form a powder to obtain chamomile extract.

[0021] According to the same method above, weigh 100 g of white willow bark to obtain white willow bark extract; weigh 100 g of centella asiatica to obtain centella asiatica extract.

[0022] Step S4: Preparation of the composition containing stem cell exosomes Mix 2 g of chamomile extract, 2 g of white willow bark extract, 2 g of centella asiatica extract, 2 g of zinc PCA, 6 g of salicylic acid, 8 g of panthenol, 2 g of sodium hyaluronate, and 4 g of exosomes by stirring for 30 minutes to prepare the composition containing stem cell exosomes.

[0023] Example 3 Preparation of a Composition Containing Stem Cell Exosomes This example provides a method for preparing a composition containing stem cell exosomes, which includes the following steps: Step S1: Cultivation of bone marrow mesenchymal stem cells Disinfect the posterior superior iliac spine at the puncture site, and after local anesthesia, aspirate 10 mL of bone marrow fluid with a bone marrow puncture needle. Inject the bone marrow fluid into a centrifuge tube containing anticoagulant, gently mix, dilute the bone marrow fluid with PBS buffer at a ratio of 1:1, filter it through a 100 μm cell sieve into a 50 mL centrifuge tube. Add 3 volumes of red blood cell lysate, let it stand at room temperature for 5 minutes, centrifuge at 300×g for 5 minutes, and discard the supernatant. Resuspend the cells with PBS buffer, centrifuge at 300×g for 5 minutes, and repeat 2 times. Resuspend the cells with 10 mL of MSCGM-CD™ medium, take 10 μL of the suspension for cell counting, and inoculate it into a T75 culture flask at a density of 5×10 5 cells / cm 2 and place it in an incubator. After culturing for 48 hours, discard the medium, gently add 5 mL of PBS buffer and wash 2 times to remove non-adherent cells, then add 10 mL of MSCGM-CD™ medium. When the cell confluence reaches 85%, discard the medium and wash 1 time with PBS buffer. Add 2 mL of 0.25% trypsin-EDTA, digest at 37°C for 5 minutes, add MSCGM-CD™ medium to terminate digestion, gently pipette to obtain a single cell suspension, centrifuge at 300×g for 5 minutes, and discard the supernatant. Resuspend with MSCGM-CD™ medium, inoculate into a new culture flask at a ratio of 1:3, and when the cell confluence reaches 65%, centrifuge at 300×g for 10 minutes and collect the supernatant. Step S2: Extraction of exosomes Transfer the supernatant to a new tube, centrifuge at 2000×g for 20 minutes to remove dead cells and debris, filter the supernatant through a 0.22 μm filter membrane to remove residual particles. Aliquot the filtrate into ultracentrifuge tubes, centrifuge at 100000×g at 4°C for 70 minutes, discard the supernatant, and retain the precipitate at the bottom of the tube. Resuspend the precipitate with pre-cooled PBS buffer, gently pipette to avoid damaging the exosomes, centrifuge at 100000×g at 4°C for 70 minutes, and discard the supernatant. Resuspend the precipitate with 200 μL of sterile PBS buffer to obtain exosomes derived from human bone marrow mesenchymal stem cells.

[0024] Step S3: Preparation of chamomile, white willow bark, and centella asiatica extracts Take 100 g of dried chamomile, crush it and sieve it through a 40-mesh sieve, weigh it and set aside. Mix the chamomile powder with 70% ethanol at a ratio of 1:15 (w / v), and soak it at room temperature for 30 minutes. Perform ultrasonic extraction, set the ultrasonic power at 300 W, the temperature at 50 °C, and the extraction time at 30 minutes. After the ultrasonic extraction, centrifuge at 4000×g for 15 minutes and collect the supernatant. Concentrate the supernatant to 1 / 5 of the original volume using a rotary evaporator under the conditions of 50 °C and a vacuum of -0.08 MPa. Freeze-dry it into a powder to obtain the chamomile extract.

[0025] According to the same method as above, weigh 100 g of white willow bark to obtain the white willow bark extract; weigh 100 g of gotu kola to obtain the gotu kola extract.

[0026] Step S4: Preparation of the composition containing stem cell exosomes Mix 1.5 g of chamomile extract, 1.5 g of white willow bark extract, 1.5 g of gotu kola extract, 1.5 g of zinc PCA, 4.5 g of salicylic acid, 6 g of panthenol, 1.5 g of sodium hyaluronate, and 3 g of exosomes by stirring for 30 minutes to prepare the composition containing stem cell exosomes.

[0027] Comparative Example 1 Compared with Example 3, the difference is that zinc PCA, chamomile extract, white willow bark extract, and gotu kola extract are not added to the prepared composition.

[0028] Comparative Example 2 Compared with Example 3, the difference is that exosomes are not added to the prepared composition.

[0029] Test Example 1 Skin Patch Test Test samples: The compositions containing stem cell extract prepared in Examples 1 - 3 and Comparative Examples 1 - 2 of the present invention were formulated into a sample solution of 1 mg / mL.

[0030] Subject screening: 30 healthy volunteers, aged 18 - 60 years old, with a balanced male-female ratio.

[0031] Experimental method: Wipe the face, upper back, or flexor side of the forearm with a 75% alcohol cotton pad and let it dry naturally. Take 15 μL of each test sample and drop it onto the filter paper of the patch tester, and fix the patch tester to the test site with medical tape (spacing ≥ 2 cm). Seal for 48 hours, and avoid strenuous exercise, getting wet, or rubbing the test site during this period. Remove the patch tester, and observe 30 minutes after removal to record immediate erythema, edema, papules and other reactions.

[0032] The skin reaction scoring criteria are shown in Table 1.

[0033] Table 1: The test results are shown in Table 2.

[0034] Table 2: As can be seen from the above table, the compositions containing stem cell extract prepared in Examples 1-3 and Comparative Examples 1-2 of the present invention have good skin mildness, no irritation to human skin, are safe to use, and have no allergic reactions.

[0035] Test Example 2 Bacteriostatic Experiment The bacteriostatic effect of the skin care lotion in the present invention on Propionibacterium acnes was used to test its bacteriostatic effect in this test.

[0036] The following experiments were set up with three replicates, and the results were averaged.

[0037] Test samples: Compositions containing stem cell extract prepared in Examples 1-3 and Comparative Examples 1-2 of the present invention were formulated into a sample solution of 1 mg / mL.

[0038] Strains: Standard strains of Propionibacterium acnes. Bacteria were taken from a -80 °C glycerol cryopreservation tube, streaked and inoculated onto an enhanced clostridial medium agar plate, and anaerobically cultured for 48 hours (37 °C). Single colonies were picked and inoculated into 5 mL of brain heart infusion medium, anaerobically cultured for 48 hours, and the bacterial solution concentration was adjusted to 1×10 6 CFU / mL. 1 mL of the bacterial solution was pipetted onto an enhanced clostridial medium agar plate, spread evenly with a spreader, and sterile Oxford cups were placed. One of the Oxford cups in a petri dish was not added with the test sample and used as a blank control group, and 200 μL of the test sample was added to the other Oxford cups respectively. Anaerobically cultured for 16 hours, the diameter of the bacteriostatic zone was measured (≥6 mm is effective). A bacteriostatic zone diameter ≤10 mm indicates no bacteriostatic effect, a bacteriostatic zone diameter ≥10 mm and ≤15 mm is moderate bacteriostasis, and a bacteriostatic zone diameter ≥15 mm is high bacteriostasis. The results are shown in Table 3.

[0039] Table 3: As can be seen from the above table, the compositions containing stem cell extract prepared in Examples 1 to 3 of the present invention have a better inhibitory effect on Propionibacterium acnes. This is because the pro-inflammatory factors carried by exosomes and bisabolol in chamomile extract form a dual anti-inflammatory mechanism. At the same time, the triterpenoid components in centella asiatica enhance the bactericidal effect by destroying the biofilm of Propionibacterium acnes, and the exosomal heat shock protein HSP70 maintains the integrity of the skin barrier. The two work together to reconstruct the skin microenvironment. While the inhibitory effects of the compositions obtained in Comparative Examples 1 to 2 on Propionibacterium acnes are weak, indicating that the composition of the present invention can effectively inhibit Propionibacterium acnes, thereby achieving antibacterial from the source and reducing the occurrence of skin inflammation problems caused by skin infections.

[0040] Test Example 3 Skin Oil Content Test Test Samples: The compositions containing stem cell extract prepared in Examples 1 to 3 and Comparative Examples 1 to 2 of the present invention were formulated into a sample solution of 1 mg / mL.

[0041] Subject Screening: 50 healthy volunteers, aged 18 - 60 years old, with a balanced male-female ratio, were randomly divided into 5 groups, with 10 people in each group.

[0042] Experimental Method: Evaluation was carried out using the German CK skin oil tester Sebumeter SM815. The subjects sat quietly in the laboratory for 30 minutes to adapt to the temperature and humidity. The forehead was washed with a fat-free facial cleanser, gently patted dry after rinsing with warm water. The skin oil content was measured in a 3 cm × 3 cm test area on the forehead of the volunteers. The oil content of the forehead was measured and recorded as the initial oil content. 0.5 mL of the test sample was applied to the fixed area of the forehead for 1 minute, and then the oil content of the three designated areas was measured. The measurement was taken every 30 minutes for a duration of 2 hours. The percentage increase in oil content was calculated for evaluation, and the results are shown in Table 4.

[0043] Table 4: As can be seen from the above table, the compositions containing stem cell extract prepared in Examples 1 to 3 of the present invention have good oil control ability. This is because zinc PCA reduces sebum secretion induced by DHT by inhibiting 5α-reductase, and regulates the PPARγ pathway with sphingosine-1-phosphate in exosomes to reduce lipid synthesis to form a complement. Salicin in white willow bark extract enhances the permeability of zinc PCA while dissolving the horny plug, making zinc ions act more effectively on the basal cells of sebaceous glands. While the oil control ability of the compositions obtained in Comparative Examples 1 to 2 is weak, indicating that the composition of the present invention can achieve the effect of regulating the skin's water-oil balance.

[0044] In summary, the present invention constructs a synergistic system of exosomes and various components, and realizes the synergy of anti-inflammatory repair, cortical regulation, and microecology through a multi-target action mechanism, thereby enhancing the antibacterial and oil-control effects of the composition.

[0045] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit them; although the present invention has been described in detail with reference to the foregoing embodiments, those of ordinary skill in the art should understand that they can still modify the technical solutions described in the foregoing embodiments, or perform equivalent replacements on some or all of the technical features; and these modifications or replacements do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. A method for preparing a composition containing stem cell exosomes, characterized in that, The following steps are involved: Step S1: Cultivation of mesenchymal stem cells: Cultivate mesenchymal stem cells with MSCGM-CD™ medium. When the cell confluence reaches 80%-90%, discard the medium, wash with PBS buffer, add 2 mL of trypsin-EDTA for digestion, terminate the digestion, gently pipette to form a single cell suspension, centrifuge and discard the supernatant, resuspend with new MSCGM-CD™ medium, inoculate into a new culture bottle, centrifuge when the cell confluence reaches 60%-70%, and collect the supernatant; Step S2: Extraction of exosomes: transfer the supernatant obtained in step S1 to a new tube, centrifuge to remove dead cells and debris, filter the supernatant, remove residual particles, dispense the filtrate into ultracentrifuge tubes, centrifuge to discard the supernatant, retain the precipitate at the bottom of the tube, resuspend the precipitate with pre-cooled PBS buffer, gently pipette, centrifuge to discard the supernatant, resuspend the precipitate with sterile PBS buffer to obtain exosomes; Step S3: Preparation of chamomile, white willow bark and Centella asiatica extracts: chamomile is taken, crushed and sieved, the obtained chamomile powder is mixed with 70% ethanol, immersed at room temperature for 30 minutes, ultrasonically extracted, centrifuged after the ultrasonication, the supernatant is collected, the supernatant is concentrated by rotary evaporator, and freeze-dried into powder to obtain chamomile extract; white willow bark extract and Centella asiatica extract are obtained respectively according to the same operation as chamomile; Step S4: Preparation of a composition containing stem cell exosomes: PCA zinc, salicylic acid, panthenol, sodium hyaluronate, the exosomes described in step S2, the chamomile extract, white willow bark extract and Centella asiatica extract described in step S3 are stirred and mixed for 30 minutes to prepare a composition containing stem cell exosomes.

2. The preparation method according to claim 1, characterized in that, The mesenchymal stem cells in step S1 are human bone marrow mesenchymal stem cells.

3. The preparation method according to claim 1, characterized in that, The concentration of trypsin-EDTA in step S1 is 2%-3%.

4. The preparation method according to claim 1, characterized in that, The mass of PCA zinc in step S4 is 1-2 g, the mass of salicylic acid is 3-6 g, the mass of panthenol is 4-8 g, the mass of sodium hyaluronate is 1-2 g, the mass of chamomile extract is 1-2 g, the mass of white willow bark extract is 1-2 g, the mass of Centella asiatica extract is 1-2 g, and the mass of exosomes is 2-4 g.

5. A composition containing stem cell exosomes prepared by the preparation method according to any one of claims 1 to 4.

6. Use of the composition containing stem cell exosomes as claimed in claim 5 in cosmetics.

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