Exosome composition and its use in improving pores

Abstract

The present application belongs to the technical field of exosome application, and discloses an exosome composition, which is obtained by mixing exosomes of Funaria hygrometrica, exosomes of Cnidium monnieri and exosomes of mesenchymal stem cells according to a mass ratio of 2-5:3-8:0.5-1. The exosomes of Funaria hygrometrica can significantly reduce the concentration of inflammatory factors, thereby achieving the effects of inhibiting excessive secretion of sebum and improving pores. The exosomes of Cnidium monnieri also synergize with the exosomes of Funaria hygrometrica to strengthen normal differentiation of keratinocytes, effectively alleviate excessive keratin layer multi-layering caused by excessive secretion of sebum, and thereby inhibit the volume increase of hair follicles. After the exosomes of mesenchymal stem cells and the plant exosomes are combined, the elasticity of the skin can be significantly improved, and the problem of skin laxity can be improved, so that the pores become more delicate, and the pores are significantly reduced in vision. The exosome composition of the present application has the effects of significantly improving coarse pores and increasing skin elasticity, can be made into various product forms, and has a wide application prospect.

Description

Technical Field

[0001] This invention belongs to the field of exosome application technology, and more specifically, relates to an exosome composition and its application in improving pores. Background Technology

[0002] Pores are the porous structures exposed on the grain of the skin after hair removal. As dead skin cells accumulate, the skin thickens and becomes rougher, leading to larger pores. The skin also becomes dull and dry because it cannot properly absorb moisture and skincare ingredients, further stimulating sebum production and causing pores to enlarge. Furthermore, as we age, the skin gradually loses elasticity, and the lack of supporting structures around the hair follicles also makes pores appear larger. An oily-on-the-outside, dry-on-the-inside skin environment can also cause pores to appear larger.

[0003] Existing research has summarized the causes of enlarged pores, mainly in three aspects: First, aging causes the skin to sag, reducing the elasticity of the pore walls and causing the pores to change from round to oval, making the pores more noticeable; Second, there is a positive correlation between sebum secretion and pore size, with sebum production exceeding 0.15 mg / cm³ every 3 hours. 2 At this time, enlarged pores may occur. Excessive sebum secretion will cause an increase in unsaturated fatty acids, resulting in incomplete keratinization around the pores, thus causing the pores to become larger. Third, the hair follicle volume increases, mainly due to abnormal differentiation of keratinocytes. At this time, the skin will become thinner and drier, causing dead skin cells to accumulate in the pores, which will enlarge the pores.

[0004] Exosomes are small membrane vesicles (30-150 nm) containing complex RNA and proteins; currently, they specifically refer to disc-shaped vesicles with a diameter of 40-100 nm. Various cells can secrete exosomes under both normal and pathological conditions. They mainly originate from multivesicles formed by the invagination of lysosomal microparticles within the cell, and are released into the extracellular matrix after the outer membrane of the multivesicle fuses with the cell membrane.

[0005] Emerging cosmetic procedures, such as electrical stimulation, can shrink pores to some extent by stimulating collagen regeneration. However, they can cause skin damage, requiring a long recovery period and numerous precautions, and are not suitable for long-term or repeated use. Mesenchymal stem cell exosomes can restore the activity of skin fibroblasts and promote collagen regeneration through various cytokines, offering powerful cosmetic effects. However, their effectiveness in improving enlarged pores is relatively poor. Summary of the Invention

[0006] The technical problem to be solved by the present invention is to overcome the above-mentioned problems existing in the prior art, and firstly, to provide an exosome composition.

[0007] A second objective of this invention is to provide the application of the above-described exosome composition in improving enlarged pores.

[0008] The objective of this invention is achieved through the following technical solution:

[0009] The present invention first provides an exosome composition, which is composed of plant exosomes and mesenchymal stem cell exosomes, wherein the plant exosomes are cucurbita exosomes and Cnidium monnieri exosomes, and the cucurbita exosomes, Cnidium monnieri exosomes and mesenchymal stem cell exosomes are mixed in a mass ratio of 2-5:3-8:0.5-1.

[0010] It is well known in the art that *Hymenochloa crus-galli* can dispel wind and dampness, relieve pain, and stop bleeding, and is mainly used to treat rheumatic pain, sinusitis, traumatic injuries, and tuberculosis with hemoptysis. However, there is no relevant research on *Hymenochloa crus-galli* exosomes. Previous research in this invention found that extracting *Hymenochloa crus-galli* exosomes can significantly reduce the concentration of inflammatory factors, thereby inhibiting excessive sebum secretion and improving pore size. Osthol has anti-inflammatory effects in many inflammatory models. Existing technologies mostly use *Cnidium monnieri* for immune-related skin diseases. In this invention, *Cnidium monnieri* exosomes retain the effect of reducing the concentration of inflammatory factors and also work synergistically with *Hymenochloa crus-galli* exosomes to enhance the normal differentiation of keratinocytes, effectively alleviate the excessive stratum corneum multiplication caused by excessive sebum secretion, thereby inhibiting the increase in hair follicle volume. As a result, the pore volume becomes smaller and the phenomenon of enlarged pores is greatly improved.

[0011] Mesenchymal stem cell exosomes have been reported to have a significant effect on skin wound repair, but there are no reports on their effect on improving pores. Here, the combination of mesenchymal stem cell exosomes and plant exosomes can significantly improve skin elasticity and improve skin laxity, thereby making pores more refined and visually shrinking them.

[0012] Therefore, the present invention also provides the use of the above-described exosome composition in improving pores.

[0013] Preferably, the exosome composition has the following functions:

[0014] (1) Shrink skin pores;

[0015] (2) Reduce skin oil secretion;

[0016] (3) Increases skin elasticity;

[0017] (4) Reduce the secretion of inflammatory factors.

[0018] The present invention also provides a method for preparing the above-mentioned exosome composition, comprising the following steps:

[0019] S1. Preparation of plant exosomes:

[0020] S11. Take *Cnidium monnieri* / *Cnidium monnieri*, add buffer solution and crush to obtain the crushed liquid;

[0021] S12. After centrifuging the lysate 2-3 times, take the supernatant to obtain exosomes of *Cnidium monnieri* and exosomes of *Cnidium monnieri*, respectively.

[0022] S2. Preparation of mesenchymal stem cell exosomes:

[0023] S21. Collect MSC cell supernatant, centrifuge 2-3 times, take the supernatant and filter it through a membrane.

[0024] S22. After ultracentrifugation of the supernatant after membrane filtration, the precipitate is collected and resuspended to obtain mesenchymal stem cell exosomes.

[0025] Preferably, the preparation of the plant exosomes includes the following steps:

[0026] (1) Take *Cnidium monnieri*, add an equal amount of PBS buffer, and use a crusher at 4°C for 10-25 min. After crushing for 1 min, stop for 1 min to obtain the lysate.

[0027] (2) Centrifuge the lysate 2 to 3 times. The first centrifugation is performed at 4℃, 6000 to 8000g for 5 to 10 minutes. Take the supernatant from the first centrifugation. Centrifuge the supernatant from the first centrifugation a second time at 4℃, 8000 to 10000g for 5 to 10 minutes. Take the supernatant from the second centrifugation. If there is a third centrifugation, the conditions for the third centrifugation are the same as those for the second centrifugation. After centrifugation, take the supernatant to obtain the exosomes of *Cnidium monnieri* / *Cnidium monnieri*.

[0028] Preferably, the preparation of the mesenchymal stem cell exosomes includes the following steps:

[0029] (1) Collect 10 fusions that have grown to 60%–80% fusion. 8 ~10 9 Centrifuge the supernatant of mesenchymal stem cells at 3000-5000g and collect the supernatant.

[0030] (2) Take the supernatant from (1), centrifuge for 3000-5000g to remove cell debris, and collect the supernatant.

[0031] (3) Take the supernatant from (2), centrifuge at 8000-10000g to remove protein, collect the supernatant, and then filter the supernatant through a 0.22μm filter membrane.

[0032] (4) Take the supernatant after filtration of (3) and centrifuge at high speed for 100,000 g for 1-2 h. Discard the supernatant, resuspend the precipitate with PBS, and collect the resuspended liquid.

[0033] (5) Take the resuspended solution from (4) to detect the content of exosomes and mesenchymal stem cell exosomes.

[0034] The present invention also provides products containing the exosome composition.

[0035] Specifically, the above-mentioned exosome composition can be combined with excipients to make a product with the effect of improving pores. The product can be a skin care water, skin care essence, skin care lotion, or it can be mixed with other media to make a skin care mask.

[0036] Preferably, the exosome composition has the following functions:

[0037] (1) Reduce the diameter of skin pores;

[0038] (2) Reduce skin oil secretion;

[0039] (3) Increases skin elasticity;

[0040] (4) Reduce the secretion of inflammatory factors.

[0041] Compared with the prior art, the present invention has the following beneficial effects:

[0042] This invention discloses an exosome composition, obtained by mixing *Hymenochloa crus-galli* exosomes, *Cnidium monnieri* exosomes, and mesenchymal stem cell exosomes in a mass ratio of 2–5:3–8:0.5–1. *Hymenochloa crus-galli* exosomes can significantly reduce the concentration of inflammatory factors, thereby inhibiting excessive sebum secretion and improving pore size. *Cnidium monnieri* exosomes, while retaining the effect of reducing inflammatory factor concentration, also synergistically work with *Hymenochloa crus-galli* exosomes to enhance the normal differentiation of keratinocytes, effectively alleviating excessive stratum corneum multiplication caused by excessive sebum secretion, thereby inhibiting the increase in hair follicle volume. The combination of mesenchymal stem cell exosomes and plant exosomes can significantly improve skin elasticity, improve skin laxity, thus making pores more refined and visually significantly shrinking them.

[0043] The exosome composition of the present invention has the effect of significantly improving enlarged pores and increasing skin elasticity, and can be made into various product forms, with broad application prospects. Attached Figure Description

[0044] Figure 1 Electron micrograph of exosomes from *Cymbidium goeringii*;

[0045] Figure 2 Electron micrograph of Cnidium monnieri exosomes;

[0046] Figure 3 Electron micrograph of mesenchymal stem cell exosomes;

[0047] Figure 4This image shows a comparison of pore improvement before and after using the exosome composition gel. Detailed Implementation

[0048] The specific embodiments of the present invention will be further described below. It should be noted that these descriptions are for the purpose of aiding understanding the present invention, but do not constitute a limitation thereof. Furthermore, the technical features involved in the various embodiments of the present invention described below can be combined with each other as long as they do not conflict with each other.

[0049] It was obtained by mixing exosomes of *Cnidium monnieri*, exosomes of *Cnidium monnieri*, and exosomes of mesenchymal stem cells in a mass ratio of 2–5:3–8:0.5–1.

[0050] Example 1

[0051] An exosome composition is obtained by mixing exosomes of *Cnidium monnieri*, exosomes of *Cnidium monnieri*, and exosomes of mesenchymal stem cells in a mass ratio of 2:3:0.5.

[0052] The preparation process of exosomes from *Cymbidium goeringii* is as follows:

[0053] (1) Take the hyacinth moss, add an equal amount of PBS buffer, and use a crusher to crush it at 4°C for 15 minutes. Crush for 1 minute and stop for 1 minute to obtain the crushed liquid.

[0054] (2) The lysate was centrifuged twice. The first centrifugation was performed at 4℃, 6000g, for 10 min. The supernatant from the first centrifugation was collected. This supernatant was then centrifuged a second time at 4℃, 10000g, for 10 min. The supernatant from the second centrifugation was collected. After testing, the exosomes of *Bryophyta cucumeroides* were obtained, and their characteristics were as follows: Figure 1 .

[0055] The preparation process of Cnidium monnieri exosomes is as follows:

[0056] (1) Take Cnidium monnieri, add an equal amount of PBS buffer, and use a crusher to crush at 4°C for 10 min. After crushing for 1 min, stop for 1 min to obtain the crushed liquid.

[0057] (2) The lysate was centrifuged three times. The first centrifugation was performed at 4℃, 6000g, for 10 min. The supernatant from the first centrifugation was collected. This supernatant was then centrifuged a second time at 4℃, 8000g, for 10 min. The supernatant from the second centrifugation was collected. This supernatant was then centrifuged a third time at 4℃, 10000g, for 15 min. The supernatant from all three centrifugations was collected. After analysis, the exosomes of *Cnidium monnieri* were obtained, and their characterization was as follows: Figure 2 .

[0058] The preparation process of mesenchymal stem cell exosomes is as follows:

[0059] (1) Collect 10 fusions that have reached 80% fusion. 8 Centrifuge the supernatant of mesenchymal stem cells at 3000g and collect the supernatant.

[0060] (2) Take the supernatant from (1), centrifuge for 3000g to remove cell debris, and collect the supernatant;

[0061] (3) Take the supernatant from (2), centrifuge for 10000g to remove protein, collect the supernatant, and then filter the supernatant through a 0.22μm filter membrane.

[0062] (4) Take the supernatant after filtration of (3) and centrifuge at high speed for 100,000 g for 1.5 h. Discard the supernatant, resuspend the precipitate with PBS, and collect the resuspended liquid.

[0063] (5) Take the resuspended solution from (4) to detect the exosome content of mesenchymal stem cell exosomes, and characterize them as follows: Figure 3 .

[0064] Carbomer powder was uniformly dispersed into the exosome composition obtained by the above mixture, and swollen at room temperature to obtain exosome composition gel 1.

[0065] Example 2

[0066] An exosome composition is obtained by mixing exosomes of *Cnidium monnieri*, exosomes of *Cnidium monnieri*, and exosomes of mesenchymal stem cells in a mass ratio of 3:5:0.8.

[0067] The preparation process of exosomes from *Cymbidium goeringii* is as follows:

[0068] (1) Take the hyacinth moss, add an equal amount of PBS buffer, and use a crusher to crush it at 4°C for 20 min. Crush for 1 min and stop for 1 min to obtain the crushed liquid.

[0069] (2) The lysate was centrifuged 3 times. The first centrifugation was performed at 4℃, 6000g, for 8 min. The supernatant of the first centrifugation was collected. The supernatant of the first centrifugation was centrifuged a second time at 4℃, 8000g, for 10 min. The supernatant of the second centrifugation was collected. The supernatant of the second centrifugation was centrifuged a third time at 4℃, 10000g, for 15 min. The supernatant of the third centrifugation was collected. After testing, the exosomes of *Cynodon dactylon* were obtained.

[0070] The preparation process of Cnidium monnieri exosomes is as follows:

[0071] (1) Take Cnidium monnieri, add an equal amount of PBS buffer, and use a crusher to crush at 4°C for 15 min. Crush for 1 min, then stop for 1 min to obtain the crushed liquid.

[0072] (2) The lysate was crushed and centrifuged twice. The first centrifugation was carried out at 4℃, 8000g, for 15min. The supernatant of the first centrifugation was taken. The supernatant of the first centrifugation was then centrifuged a second time at 4℃, 10000g, for 15min. The supernatant of the second centrifugation was taken and tested to obtain Cnidium monnieri exosomes.

[0073] The preparation process of mesenchymal stem cell exosomes is as follows:

[0074] (1) Collect 10 fusions that have reached 80% fusion. 8 Centrifuge the supernatant of mesenchymal stem cells at 3000g and collect the supernatant.

[0075] (2) Take the supernatant from (1), centrifuge for 3000g to remove cell debris, and collect the supernatant;

[0076] (3) Take the supernatant from (2), centrifuge for 10000g to remove protein, collect the supernatant, and then filter the supernatant through a 0.22μm filter membrane.

[0077] (4) Take the supernatant after filtration of (3) and centrifuge at high speed for 100,000 g for 2 h. Discard the supernatant, resuspend the precipitate with PBS, and collect the resuspended liquid.

[0078] (5) Take the resuspended solution from (4) to detect the content of exosomes and mesenchymal stem cell exosomes.

[0079] Carbomer powder was uniformly dispersed into the exosome composition obtained by the above mixture, and swollen at room temperature to obtain exosome composition gel 2.

[0080] Example 3

[0081] An exosome composition is obtained by mixing exosomes of *Cnidium monnieri*, exosomes of *Cnidium monnieri*, and exosomes of mesenchymal stem cells in a mass ratio of 5:8:1.

[0082] The preparation of exosomes from *Cnidium monnieri*, exosomes from *Cnidium monnieri*, and exosomes from mesenchymal stem cells was carried out according to Example 2.

[0083] Carbomer powder was uniformly dispersed into the exosome composition obtained by the above mixture, and swollen at room temperature to obtain exosome composition gel 3.

[0084] Comparative Example 1

[0085] An exosome composition is obtained by mixing moss exosomes and mesenchymal stem cell exosomes in a mass ratio of 5:1.

[0086] The preparation of exosomes from *Calamus hupensis* and exosomes from mesenchymal stem cells was carried out according to Example 2.

[0087] Carbomer powder was uniformly dispersed into the exosome composition obtained by the above mixture, and swollen at room temperature to obtain exosome composition gel 4.

[0088] Comparative Example 2

[0089] An exosome composition is obtained by mixing Cnidium monnieri exosomes and mesenchymal stem cell exosomes at a mass ratio of 8:1.

[0090] The preparation of Cnidium monnieri exosomes and mesenchymal stem cell exosomes was carried out according to Example 2.

[0091] Carbomer powder was uniformly dispersed into the exosome composition obtained by the above mixture, and swollen at room temperature to obtain exosome composition gel 5.

[0092] Comparative Example 3

[0093] An exosome composition is obtained by mixing exosomes of *Cynodon dactylon* and exosomes of *Cnidium monnieri* in a mass ratio of 5:8.

[0094] The preparation of exosomes from *Cnidium monnieri* and *Cnidium monnieri* was carried out according to Example 2.

[0095] Carbomer powder was uniformly dispersed into the exosome composition obtained by the above mixture, and swollen at room temperature to obtain exosome composition gel 6.

[0096] Comparative Example 4

[0097] An exosome composition is obtained by mixing exosomes of *Cnidium monnieri*, exosomes of *Cnidium monnieri*, and exosomes of mesenchymal stem cells in a mass ratio of 5:8:0.5.

[0098] The preparation of exosomes from *Cnidium monnieri*, exosomes from *Cnidium monnieri*, and exosomes from mesenchymal stem cells was carried out according to Example 2.

[0099] Carbomer powder was uniformly dispersed into the exosome composition obtained by the above mixture, and swollen at room temperature to obtain exosome composition gel 7.

[0100] Test case

[0101] I. Safety Test

[0102] 1. Experimental animals: Male SPF 6-week-old mice.

[0103] 2. Experimental Design: Eight healthy mice with intact skin were selected and divided into four groups. After routine anesthesia, the abdominal hair was removed, covering an area of ​​2cm × 2cm. 0.2g of the exosome composition gel prepared in Examples 1 to 3 was applied to the hairless skin surface of each group. The control group mice received a blank gel. After 12 hours, the skin at the application site was washed with warm water, and the skin reaction was observed. The results showed that, compared with the control group, no redness or inflammation was observed on the skin surface of the experimental animals, indicating that the exosome composition gel prepared in this invention is non-irritating to the skin of the experimental animals.

[0104] II. Inflammatory Factor Detection

[0105] The skin of mice that had been coated with the exosome composition gel in the above experiment was removed, homogenized, and used for the detection of immune inflammatory factors. The specific detection procedure was performed according to the existing kit.

[0106] The results are shown in Table 1. Table 1 shows that the specific implementation methods of the technical solution adopted in this invention (Examples 1 to 3) all have a significant effect on reducing IL-1α. Comparison of data from Comparative Examples 1, 2, and 3 clearly shows that the synergistic effect of *Cnidium monnieri* exosomes and *Brucea javanica* exosomes can further reduce the expression of inflammatory factors. Data from Comparative Example 3 indicates that mesenchymal stem cell exosomes have a relatively small impact on inflammatory factors; data from Comparative Example 4 shows that the ratio of the exosome composition has a relatively small impact on inflammatory factors.

[0107] In addition, Table 1 also lists the changes in the levels of inflammatory factors IL-4 and IL-5. The production of some immune inflammatory factors such as IL-4 and IL-5 exacerbates skin reactions, thereby leading to damage to the skin barrier. Table 1 shows the changes in the levels of inflammatory factors IL-4 and IL-5. The exosome composition of the present invention can also reduce the levels of inflammatory factors IL-4 and IL-5, thereby protecting the skin barrier, avoiding skin irritation, and thus avoiding problems such as rough skin and enlarged pores caused by damage to the skin barrier.

[0108] Table 1

[0109] Grouping IL-1α (pg / mL) IL-4 (pg / mL) IL-5 (pg / mL) Example 1 64.84±0.33 3.73±0.24 2.81±0.27 Example 2 65.62±0.24 3.64±0.26 2.76±0.29 Example 3 62.71±0.45 3.45±0.22 2.69±0.36 Comparative Example 1 82.85±0.33 4.86±0.35 3.85±0.16 Comparative Example 2 73.06±0.27 4.31±0.29 3.29±0.24 Comparative Example 3 65.27±0.19 5.57±0.28 4.02±0.15 Comparative Example 4 65.19±0.28 5.61±0.31 4.28±0.16 Blank group 95.38±0.31 5.87±0.16 4.46±0.22

[0110] III. Skin Change Detection

[0111] Fifty-four eligible volunteers were selected, half of whom were healthy women and half were healthy men. They had severe facial skin pores, with a pore grade of 3-4 (based on the Asian Skin Aging Atlas - Volume 2 Asian Type). They had no history of cosmetic allergies and were aged between 30 and 39 years old.

[0112] Volunteers were divided into 9 groups, with 3 males and 3 females in each group. Volunteers used the exosome gel compositions of Examples 1 to 3 and Comparative Examples 1 to 4. A positive control group (using a competing product on the market) and a blank control group (blank gel) were set up. The number of pores around the cheeks and nose were calculated using image analysis. The results of the test after 30 days, including the reduction in the number of pores, pore diameter and volume, were analyzed. The experimental results of the blank control group were used as the standard. The calculation results are shown in Table 2.

[0113] As shown in Table 2, compared with the comparative examples and the blank group, the exosome composition of the present invention can significantly reduce the number and volume of pores, thereby significantly improving enlarged pores and achieving a good pore-shrinking effect. A comparison of the data from Comparative Examples 1, 2, and 3 clearly shows that the synergistic effect of *Cnidium monnieri* exosomes and *Lysimachia christinae* exosomes significantly improves the problem of enlarged pores. The data from Comparative Example 3 indicates that the lack of animal exosomes has a relatively small impact on pore improvement. The data from Comparative Example 4 shows that the ratio of the exosome composition is crucial in improving pores. The inventors hypothesize that the interaction between *Cnidium monnieri* exosomes, *Lysimachia christinae* exosomes, and mesenchymal stem cell exosomes produces factors that affect the working mechanism of skin cells.

[0114] Table 2

[0115] Grouping Pore ​​reduction rate (%) Pore ​​volume reduction rate (%) Pore ​​improvement rate (%) Example 1 24.13±0.35 18.69±0.21 39.64±0.28 Example 2 25.65±0.28 17.39±0.26 38.84±0.29 Example 3 32.37±0.42 23.14±0.23 42.86±0.34 Comparative Example 1 6.49±0.31 8.87±0.39 13.81±0.18 Comparative Example 2 6.23±0.27 8.36±0.26 13.29±0.29 Comparative Example 3 5.36±0.26 5.49±0.28 4.08±0.14 Comparative Example 4 3.17±0.13 3.97±0.35 3.27±0.19 Blank group 0.12±0.03 2.89±0.17 1.47±0.25

[0116] Sebum tests were performed on the forehead on days 10, 20, and 30 of the test, and the results are shown in Table 3. As can be seen from Table 3, the amount of oil decreased more and more significantly over time in the example groups, especially the exosome composition gel of Example 3, which showed a very good oil-controlling effect. Data from Comparative Examples 1 and 2 indicate that, in the exosome composition, *Cnidium monnieri* exosomes contributed more to the oil-controlling effect of the composition gel than *Cnidium monnieri* exosomes. Furthermore, mesenchymal stem cell exosomes contributed the least to the oil-controlling effect of the composition gel. Data from Comparative Example 4 shows that the formulation of the exosome composition is very important in the oil-controlling effect.

[0117] Table 3

[0118]

[0119]

[0120] Furthermore, after 30 days of use, the elasticity of the volunteers' skin was tested using a skin elasticity tester. The results showed that compared with the blank control group, the elasticity R2 of the subjects' cheek skin was significantly increased (P < 0.05). After using the exosome composition gel of Examples 1 to 3, the elasticity R2 value of the cheek skin increased by 8.46%, 9.36%, and 11.21%, respectively. After using the exosome composition gel of Comparative Examples 1 to 3, the elasticity R2 value of the cheek skin increased by 2.38%, 3.36%, and 1.29%, respectively. It can be seen that in this invention, the combination of mesenchymal stem cell exosomes and plant exosomes can significantly improve skin elasticity, improve skin laxity, thereby making pores more refined and visually significantly reducing pore size.

[0121] IV. Application Examples

[0122] Ten female volunteers aged 30-35 with similar skin types were selected and grouped into groups of ten to test the product of Example 3. The test lasted for 28 days. The product was used once daily, morning and evening, after facial cleansing. No other skincare products were used during the test, and the dosage was approximately the same for each group. After 28 days of continuous use, three volunteers were selected, and the results are as follows: Figure 4 .

[0123] from Figure 4 It can be seen that after use, the skin's water-oil balance is better, and the enlarged pores are significantly improved.

[0124] The embodiments of the present invention have been described in detail above, but the present invention is not limited to the described embodiments. For those skilled in the art, various changes, modifications, substitutions, and variations can be made to these embodiments without departing from the principles and spirit of the present invention, and these variations still fall within the protection scope of the present invention.

Claims

1. An exosome composition, characterized in that, The exosome composition comprises plant exosomes and mesenchymal stem cell exosomes, wherein the plant exosomes are *Cnidium monnieri* exosomes and *Cnidium monnieri* exosomes, and the *Cnidium monnieri* exosomes, *Cnidium monnieri* exosomes, and mesenchymal stem cell exosomes are mixed in a mass ratio of 2:3:0.5; or, The exosomes of *Cnidium monnieri*, exosomes of *Cnidium monnieri*, and exosomes of mesenchymal stem cells were mixed in a mass ratio of 3:5:0.8; or, The exosomes of *Cnidium monnieri*, exosomes of *Cnidium monnieri*, and exosomes of mesenchymal stem cells were obtained by mixing them in a mass ratio of 5:8:

1. The preparation of the plant exosomes includes the following steps: (1) Take moss or cnidium, add an equal amount of PBS buffer, and use a crusher to crush at 4°C for 10-25 min. After crushing for 1 min, stop for 1 min to obtain the crushed liquid; (2) The lysate is centrifuged 2 to 3 times. The first centrifugation is performed at 4℃, 6000 to 8000 g, for 5 to 10 minutes. The supernatant of the first centrifugation is collected. The supernatant of the first centrifugation is then centrifuged a second time at 4℃, 8000 to 10000 g, for 5 to 10 minutes. The supernatant of the second centrifugation is collected. If there is a third centrifugation, the conditions for the third centrifugation are the same as those for the second centrifugation. The supernatant collected after centrifugation is used to obtain the exosomes of *Cnidium monnieri* or *Cnidium monnieri*. The preparation of the mesenchymal stem cell exosomes includes the following steps: (1) Collect 10 cells that have grown to 60%–80% fusion. 8 ~10 9 Centrifuge the supernatant of mesenchymal stem cells at 3000-5000g and collect the supernatant. (2) Take the supernatant from (1), centrifuge for 3000-5000g to remove cell debris, and collect the supernatant; (3) Take the supernatant from (2), centrifuge at 8000-10000g to remove protein, collect the supernatant, and then filter the supernatant through a 0.22μm filter membrane; (4) Take the supernatant after filtration of (3) and centrifuge at high speed for 100,000g for 1-2 hours. Discard the supernatant, resuspend the precipitate with PBS, and collect the resuspended liquid. (5) Take the resuspended solution from (4) and detect the exosome content to obtain mesenchymal stem cell exosomes.

2. The use of the exosome composition of claim 1 in the preparation of products for improving enlarged pores.

3. The application according to claim 2, characterized in that, The exosome composition has the following functions: (1) Shrink skin pores; (2) Reduce sebum secretion; (3) Increases skin elasticity; (4) Reduce the secretion of inflammatory factors.

4. A product containing the exosome composition of claim 1.

5. The product according to claim 4, characterized in that, The exosome composition has the following functions: (1) Shrink skin pores; (2) Reduce sebum secretion; (3) Increases skin elasticity; (4) Reduce the secretion of inflammatory factors.

Images