Gel loaded with exosome composition and application thereof

Abstract

The invention belongs to the technical field of exosome application, and discloses gel loaded with an exosome composition and application of the gel. The specific combination of the three exosomes finds that the exosomes have a remarkable promoting effect on proliferation and migration of human immortalized keratinocytes, and later detection finds that the exosomes also have a remarkable promoting effect on FLG expression quantity of the cells and have a remarkable reducing effect on proinflammatory factors of the cells; therefore, it can be determined that the exosome composition can be used for repairing damaged barriers of cells. In the application level, the exosome composition and the freeze-drying protective agent are mixed for use, the exosome composition is prepared into freeze-dried powder, the freeze-dried powder is prepared into the gel preparation convenient for skin use during use, and the application effect of the gel preparation is verified through a constructed mouse model. The result shows that the prepared gel preparation containing the exosome composition has the effect of remarkably repairing skin barrier damage after light damage, and is expected to be widely applied.

Description

Technical Field

[0001] This invention belongs to the field of exosome application technology, and more specifically, relates to a gel loaded with an exosome composition and its application. Background Technology

[0002] The skin barrier, especially the stratum corneum, the outermost layer of the epidermis, is the first line of physical, chemical, and biological defense between the human body and the external environment. Its structural integrity is crucial for maintaining skin homeostasis, preventing excessive moisture loss, and resisting external irritants, allergens, and microbial invasion. In recent years, with increasing environmental stress, the prevalence of improper skincare practices, and the rising incidence of related skin diseases, skin barrier damage has become a research hotspot and a common clinical problem in dermatology.

[0003] Exosomes, extracellular vesicles with a diameter of approximately 30-150 nanometers secreted by cells, have become a research frontier in dermatology and regenerative medicine in recent years. They carry bioactive substances such as proteins, lipids, mRNA, miRNA, and circRNA from their source cells, which can be taken up by recipient cells, thereby precisely regulating the biological functions of target cells and showing unique application potential in skin repair, regeneration, and rejuvenation. The function of exosomes exhibits "homing effect" and "content dependence," and their biological effects largely depend on the type and state of their parent cells. Currently, exosomes used in skin improvement research mainly originate from the following cell types: 1. Mesenchymal stem cells, which are currently the most widely researched and applied source. They are generally obtained from adipose tissue, bone marrow, and umbilical cord; 2. Skin-derived adult cells, which exhibit greater "tissue specificity," and their signals are more easily recognized and responded to by skin cells; 3. Plant-derived extracellular vesicles, such as those from grapes, ginger, broccoli, ginseng, apples, and other fruits and plants; these extracellular vesicles have good biocompatibility and low immunogenicity. It is rich in plant-specific antioxidants (such as polyphenols and flavonoids) and bioactive molecules. It primarily exhibits excellent antioxidant and anti-inflammatory capabilities, making it useful against UV-induced oxidative damage and skin inflammation, and has gained attention in "pure beauty" and green cosmetic formulations. However, the isolation and purification of these animal-derived cells are difficult, costly, and result in small cell numbers. Furthermore, some applications involve ethical considerations, limiting their widespread use in skin improvement, which is currently mostly in laboratory research or small-scale applications.

[0004] Exosome gels, as an innovative delivery system and dosage form, combine the biological activity of exosomes with the physical properties of gel materials, and are currently a hot topic in translational applications. Liquid exosome formulations are prone to loss from the skin surface and have unstable penetration efficiency. A gel matrix can "anchor" them to the application site, forming a "drug reservoir," slowly and continuously releasing exosomes, greatly prolonging their duration of action and improving bioavailability. This is crucial for wound repair and anti-aging applications requiring sustained action. However, the preparation process of exosome gel formulations is complex and technically demanding. Uniformly and stably loading exosomes into the gel matrix without compromising their integrity and activity requires extremely high process precision. Parameters such as mixing temperature, pH, and shear force can all affect exosome activity. Summary of the Invention

[0005] In view of the above-mentioned defects in the existing technology, the present invention firstly provides an exosome composition for skin barrier damage.

[0006] The objective of this invention is achieved through the following technical solution:

[0007] An exosome composition comprising umbilical cord mesenchymal stem cell exosomes, honeysuckle exosomes, and green tea exosomes in a weight ratio of 2-3:0.5-0.8:1-2.

[0008] The umbilical cord mesenchymal stem cell exosomes, honeysuckle exosomes, and green tea exosomes mentioned above in this invention were all prepared using conventional methods in the art.

[0009] When the above-mentioned exosome composition was applied to human immortalized keratinocytes, it was found to significantly promote cell proliferation and migration, and the expression level of FLG in the cells was significantly increased. The epidermal cell layer is the most important cell layer of the skin barrier, and the proliferative and migratory activities of epidermal cells are directly related to the skin barrier's repair capacity after damage. Therefore, the exosome composition of the present invention has the potential to be developed into skin care products or pharmaceuticals that promote skin barrier repair.

[0010] This invention also protects the use of the above-described exosome composition in the development of products that alleviate and / or treat skin barrier damage.

[0011] Preferably, the exosome composition has at least one of the following application effects:

[0012] (1) Promotes the proliferation and / or migration of immortalized human keratinocytes;

[0013] (2) Promotes the expression of FLG protein in immortalized human keratinocytes;

[0014] (3) Inhibits the expression of inflammatory factors in human immortalized keratinocytes.

[0015] The present invention also protects the lyophilization protectant of the above exosome composition, wherein the lyophilization protectant is composed of 0.3-0.5% v / v ceramide, lysine and 1.8-1.5% v / v glycerol, wherein the final concentration of lysine after mixing the exosome lyophilization protectant and exosomes is 70-80 mM.

[0016] This invention, based on previously developed freeze-drying protectants, adjusts the formulation by increasing the glycerol content. Glycerol helps protect the integrity of exosomes and also moisturizes and protects the skin barrier. Additionally, the ceramide content is increased. Ceramides reduce inflammation and protect exosomes from aggregation and membrane fusion. The resulting freeze-drying protectant maximizes the protection of the bioactivity of the exosome composition while also repairing the skin barrier.

[0017] Therefore, the present invention also protects the use of the exosome composition and lyophilization protectant in the preparation of products that alleviate and / or treat skin barrier damage.

[0018] The present invention also protects gel formulations containing the said exosome composition and lyophilization protectant.

[0019] This invention also protects the use of the gel formulation in the preparation of products that alleviate and / or treat impaired skin barrier function.

[0020] This invention explores the repair effect of the prepared gel formulation on a mouse model of photodamage by constructing a photodamage mouse model. The results show that the gel formulation prepared in this invention has a significant effect on repairing the skin barrier damage caused by photodamage.

[0021] Therefore, preferably, in the above applications, the skin barrier damage refers to skin barrier damage caused by ultraviolet radiation.

[0022] More preferably, in the above applications, the skin barrier damage caused by ultraviolet radiation is manifested as an increase in TWEL value and a decrease in stratum corneum moisture content.

[0023] Compared with the prior art, the present invention has the following beneficial effects:

[0024] This invention, through a specific combination of three exosomes, revealed a significant promoting effect on the proliferation and migration of immortalized human keratinocytes. Further testing showed a significant promoting effect on FLG expression in these cells and a significant reduction in pro-inflammatory factors, thus confirming that this exosome composition can be used to repair damaged cellular barriers. In terms of application, this invention uses a mixture of the exosome composition and a lyophilization protectant to prepare the exosome composition into a lyophilized powder. This powder is then formulated into a gel for easy skin application. The efficacy of the gel formulation was verified using a constructed mouse model. The results showed that the gel formulation containing the exosome composition prepared in this invention significantly repairs skin barrier damage caused by photodamage and holds promise for widespread application. Attached Figure Description

[0025] Figure 1 Electron micrograph of mesenchymal stem cell exosomes;

[0026] Figure 2 Electron micrograph of honeysuckle exosomes;

[0027] Figure 3 Electron micrograph of exosomes from green tea;

[0028] Figure 4 The effect of exosome composition on cell migration;

[0029] Figure 5 Bar chart showing the scores of mice in each group. Detailed Implementation

[0030] To better illustrate the purpose, technical solution, and advantages of this invention, the invention will be further described below with reference to specific drawings and embodiments. Unless otherwise specified, the experimental methods used in the embodiments are conventional methods, and the materials and reagents used are commercially available unless otherwise specified.

[0031] The exosome extraction and purification kit (cell supernatant) (Umibio: UR52121) was purchased from Shanghai Yumeibo Biotechnology Co., Ltd.

[0032] Example 1: Preparation of exosomes

[0033] I. Preparation and Characterization of Umbilical Cord Mesenchymal Stem Cell Exosomes

[0034] Includes the following steps:

[0035] (1) Collect the supernatant of mesenchymal stem cells that have grown to 80% confluence, centrifuge at 3000g, and collect the supernatant;

[0036] (2) Take the supernatant from (1), centrifuge for 3000g to remove cell debris, and collect the supernatant;

[0037] (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;

[0038] (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.

[0039] (5) Take the resuspension from (4) to detect the exosome content of mesenchymal stem cell exosomes, and perform electron microscopy as follows: Figure 1 The NTA particle size was concentrated at 102.3±28.5 nm, and the exosome protein content was determined to be 7.26 mg / mL using the BCA method.

[0040] II. Preparation and Characterization of Honeysuckle Exosomes

[0041] The procedure includes the following steps: Weigh 100 g of the required dried honeysuckle flowers and soak them in 800 mL of sterile PBS for half an hour. Homogenize using a high-speed homogenizer, filter through gauze to remove larger solid impurities and residues, and incubate overnight at 4°C. Centrifuge at 10,000 rpm for 1 hour at 4°C, collect the supernatant, and repeat the above operation 3-5 times until no visible precipitate is found. The centrifuged sample is then passed through an AKTA fluxS tangential flow filtration system, first through a 200 nm hollow fiber column, collecting the coarse extract with a particle size ≤200 nm. The coarse extract is then passed through the AKTA fluxS tangential flow filtration system again, this time through a 750 kD hollow fiber column, collecting the sample with a molecular size >750 kD, i.e., honeysuckle exosomes.

[0042] Electron microscopy of honeysuckle exosomes obtained as follows Figure 2 The NTA particle size was concentrated at 92.5±12.6 nm, and the exosome protein content was determined to be 3.15 mg / mL using the BCA method.

[0043] III. Preparation and Characterization of Green Tea Exosomes

[0044] Includes the following steps:

[0045] (1) Wash fresh green tea leaves and juice them using a low-speed screw press at a stirring speed of 40 rpm to obtain liquid juice;

[0046] (2) The juice was filtered through a 400-mesh screen to remove large suspended matter. The filtered juice was centrifuged at 10,000 × g for 10 min at 4°C to remove proteins and cell walls;

[0047] (3) The supernatant obtained by the above operation is filtered through a filter membrane with 0.22 μm pores to obtain the first filtrate;

[0048] (4) Centrifuge the first filtrate at 2000×g for 10 min and filter it through a filter membrane with 0.22μm pores to obtain the second filtrate;

[0049] (5) Centrifuge the second filtrate at 10000×g for 10 min for the first time and at 20000×g for 10 min for the second time, and then recover the supernatant;

[0050] (6) The supernatant was centrifuged at 100,000 × g for 70 min at 4 °C to obtain a precipitate. The obtained precipitate was resuspended in PBS to obtain green tea exosomes.

[0051] Electron microscopy of the obtained green tea exosomes is as follows: Figure 3 The NTA particle size was concentrated at 122.5±12.6 nm, and the exosomal protein content was determined to be 5.63 mg / mL using the BCA method.

[0052] Example 2 Cell Experiment

[0053] I. Cytotoxicity Evaluation

[0054] 100 μL / well of HaCaT cells in good growth condition were added at 5 × 10⁻⁶ cells / well. 4 Cells were seeded at a density of [number] cells / mL in 96-well plates, grouped and labeled, with three replicates per group. The plates were then incubated for 24 hours to allow for cell adhesion. The culture medium was removed, and mesenchymal stem cell exosomes, honeysuckle exosomes, and green tea exosomes were added to each well (the amount of exosomes in each well is shown in Table 1). After 24 hours of interaction with the cells, 10 μL of CCK-8 solution was added to each well, mixed thoroughly in the dark, and incubated for 1 hour. The OD value at 450 nm was measured using a microplate reader, and the cell proliferation rate was calculated.

[0055] Cell proliferation rate % = (As-Ab) / (Ac-Ab) × 100%.

[0056] Experimental group (As) = cells + culture medium + CCK-8 + exosomes;

[0057] Control group (Ac) = cells + culture medium + CCK-8;

[0058] Blank group (Ab) = culture medium + CCK-8.

[0059] Table 1

[0060] As can be seen from the data in Table 1, all concentrations of mesenchymal stem cell exosomes have a significant effect on cell proliferation, while higher concentrations of honeysuckle exosomes and green tea exosomes have a more significant inhibitory effect. Therefore, when using them, attention should be paid to the concentration combination of honeysuckle exosomes and green tea exosomes.

[0061] II. Cell proliferation and migration experiments

[0062] 1. Proliferation Experiment

[0063] The proliferation activity of HaCaT cells was measured using the conventional MTT assay, and the cells were divided into a blank control group and an experimental group.

[0064] HaCaT cells in good logarithmic growth phase were selected and their concentration adjusted to 1×10⁻⁶. 5 Cells were inoculated at a concentration of 100 μL / mL into 96-well plates, with 3 replicates per group. After complete cell adhesion, 10% of the total culture medium volume containing different concentrations of exosomes was added to the experimental group, while the blank control group contained no exosomes in its culture medium. After culturing for 24 hours, 20 μL of 5 mg / mL MTT solution was added to each well, and the cells were incubated for 4 hours. The supernatant was discarded, and 150 μL of DMSO was added and gently shaken. The absorbance (OD value) at 490 nm was measured using a microplate reader. The proliferation activity of the experimental group was calculated using the following formula, with the proliferation activity of the blank control group considered as 100%: Proliferation activity of experimental group = OD value of experimental group / OD value of blank control group × 100%. Higher absorbance indicates stronger proliferation activity.

[0065] The experimental groups are as follows:

[0066] Experimental group 1: Mesenchymal stem cell exosomes: honeysuckle exosomes: green tea exosomes = 2:0.2:1.

[0067] Experimental group 2: Mesenchymal stem cell exosomes: honeysuckle exosomes: green tea exosomes = 2:0.5:1.

[0068] Experimental group 3: Mesenchymal stem cell exosomes: honeysuckle exosomes: green tea exosomes = 2:0.8:1.

[0069] Experimental group 4: Mesenchymal stem cell exosomes: honeysuckle exosomes: green tea exosomes = 3:0.2:1.

[0070] Experimental group 5: Mesenchymal stem cell exosomes: honeysuckle exosomes: green tea exosomes = 3:0.5:1.

[0071] Experimental group 6: Mesenchymal stem cell exosomes: honeysuckle exosomes: green tea exosomes = 3:0.8:1.

[0072] Experimental group 7: Mesenchymal stem cell exosomes: honeysuckle exosomes: green tea exosomes = 2:0.2:2.

[0073] Experimental group 8: Mesenchymal stem cell exosomes: honeysuckle exosomes: green tea exosomes = 2:0.5:2.

[0074] Experimental group 9: Mesenchymal stem cell exosomes: honeysuckle exosomes: green tea exosomes = 2:0.8:2.

[0075] Experimental group 10: Mesenchymal stem cell exosomes: honeysuckle exosomes: green tea exosomes = 3:0.2:2.

[0076] Experimental group 11: Mesenchymal stem cell exosomes: honeysuckle exosomes: green tea exosomes = 3:05:2.

[0077] Experimental group 12: Mesenchymal stem cell exosomes: honeysuckle exosomes: green tea exosomes = 3:0.8:2.

[0078] Table 2

[0079] Table 2 shows that a certain mixing ratio of the three types of exosomes can significantly promote cell proliferation activity. Furthermore, compared with group 1 (group 4), group 2 (group 5), and group 3 (group 6), the increased content of mesenchymal stem cell exosomes in experimental group 4 significantly enhanced and promoted cell proliferation activity. Additionally, the changes from group 1 to group 3, group 4 to group 6, group 7 to group 9, and group 10 to group 12 indicate that the increased content of honeysuckle exosomes also promoted cell proliferation activity to some extent. Compared with group 1 (group 7), group 2 (group 8), and group 3 (group 9), the content of green tea exosomes had no significant effect on cell proliferation activity.

[0080] 2. Transfer Experiment

[0081] The migration activity of HaCaT cells was determined using the conventional scratch assay. Cells were divided into a blank control group and an experimental group. Grouping was the same as in the proliferation assay.

[0082] The results are as follows Figure 4 It can be seen that the effects of different experimental groups on cell migration and cell proliferation are consistent. The exosome compositions of the above-mentioned experimental groups in this invention can all significantly promote cell migration.

[0083] The epidermal cell layer is the most important cellular layer of the skin barrier, and the proliferative and migration activities of epidermal cells are directly related to the skin barrier's repair capacity after damage. The above experiments show that the exosome composition can more effectively enhance the proliferative activity of human immortalized epidermal HaCaT cells, and also more effectively enhance their migration activity, demonstrating promise for development into skincare products or pharmaceuticals that promote skin barrier repair.

[0084] III. Effects of Cell Barrier Protein Expression

[0085] HaCaT cells were cultured in 1640 medium containing 10% fetal bovine serum. When confluence was >90%, the cells were gently washed once or twice with PBS. Different exosome compositions (10 μL) were added to the experimental groups, while the control group received the same amount of medium. Cells were cultured at 37 °C for 48 h. After culture, cells were collected, total RNA was extracted from each experimental group, cDNA was synthesized, and q-PCR was used to detect the expression of β-actin and the target gene (the detection method was standard in the field and will not be described here). The results are shown in Table 3.

[0086] Table 3

[0087] The regulation of FLG gene expression is a key aspect of skin barrier repair. Studies have shown that upregulating FLG gene expression can enhance skin barrier function, thereby repairing damaged skin. Table 3 shows that the exosome composition of this invention can significantly increase FLG expression, thus enabling its use in the development of products for skin barrier damage. Furthermore, when the ratio of honeysuckle exosomes used is 0.2, the resulting exosome composition does not significantly promote FLG expression.

[0088] IV. Effects on inflammatory factors in damaged cells

[0089] HaCaT cells were cultured in 1640 medium containing 10% fetal bovine serum. When the confluence was >90%, the UVA / UVB ratio was 70 mJ / cm². 2 Cells were irradiated for 10 min, and exosome compositions (10 μL) were added to different experimental groups, while the control group received an equal volume of culture medium. Cells were cultured at 37℃ for 48 h. The culture medium was collected, and inflammatory factors IL-1β, IL-6, and TNF-α were detected using an ELISA kit. Inflammatory factor inhibition rate = (Inflammatory factor content in sample - Inflammatory factor content in control group) / Inflammatory factor content in control group × 100%.

[0090] The dynamic balance between keratinocyte proliferation and apoptosis is a core mechanism for maintaining skin barrier function. When this homeostasis is disrupted, the integrity of the epidermal barrier is compromised, potentially becoming a significant trigger for various inflammatory skin diseases. In a damaged skin barrier state, keratinocytes, acting as the first line of defense, work in conjunction with macrophages, dendritic cells, and T lymphocytes to activate the innate immune response. During this process, damage-associated molecular patterns (DAMPs) released from damaged tissue trigger an inflammatory cascade, significantly upregulating the expression of pro-inflammatory cytokines such as TNF-α, IL-1β, and IL-6. Table 4 shows that a certain mixing ratio of the three types of exosomes can significantly inhibit the expression of inflammatory factors. Compared with experimental group 1, experimental group 4, experimental group 5, and experimental group 2, and experimental group 6, experimental group 6 showed a slight improvement in the inhibition of inflammatory factors with increased content of mesenchymal stem cell exosomes, but the difference was not significant. Compared with experimental group 1, experimental group 1, experimental group 2, and experimental group 3, experimental group 7, experimental group 8, and experimental group 2, and experimental group 9, experimental group 9, we can also see that increased content of green tea exosomes can significantly promote the inhibition of inflammatory factor expression. In addition, when the ratio of honeysuckle exosomes is 0.2, the inhibitory effect of the resulting exosome composition on the expression of inflammatory factors is not significant.

[0091] Table 4

[0092] Example 3: Preparation of a gel formulation containing an exosome composition and mouse experiments.

[0093] I. Preparation of Gel Formulations Containing Exosome Compositions

[0094] An exosome composition and an exosome protectant are mixed to obtain a mixture, wherein the exosome composition is obtained by mixing umbilical cord mesenchymal stem cell exosomes, honeysuckle exosomes, and green tea exosomes in a weight ratio of 2-3:0.5-0.8:1-2. The exosome protectant consists of 0.3-0.5% v / v ceramide, lysine, and 1.8-1.5% v / v glycerol, wherein the final concentration of lysine in the mixture is 70-80 mM.

[0095] The mixture was freeze-dried using vacuum freeze-drying technology to obtain freeze-dried powder. The freeze-dried powder was then directly dissolved in water and mixed with hyaluronic acid and carbomer 940 to obtain a gel formulation 1 containing exosome composition.

[0096] Gel Formulation 2: Directly mix the exosome composition with hyaluronic acid and carbomer 940.

[0097] Gel Formulation 3: The mixture is freeze-dried using vacuum freeze-drying technology to obtain freeze-dried powder. After being stored for 7 days, the freeze-dried powder is dissolved in water and then mixed with hyaluronic acid and carbomer 940.

[0098] II. Mouse Experiment

[0099] In this experimental example, the exosome composition in the gel formulation was obtained according to the following weight ratio: umbilical cord mesenchymal stem cell exosomes: honeysuckle exosomes: green tea exosomes.

[0100] Mice were acclimatized for one week after purchase in an SPF laboratory animal facility. They were then randomly divided into four groups: a normal control group (CTL), an acute photodamage model group (MOD), gel preparation group 1 (N1), gel preparation group 2 (N2), gel preparation group 3 (N3), and a positive control group (Y), with nine mice in each group. The CTL group underwent no treatment except for shaving. The other groups were shaved on the back and then subjected to UVB irradiation. The MOD group received direct UVB irradiation. For the other groups, 0.2 g of the gel preparation was evenly applied to a 2.5 cm × 2.0 cm area on the back of each mouse 20 minutes before UVB irradiation. The Y group received 0.3 g of the positive control agent (Anessa sunscreen). Mice were given 1000 mJ·cm² daily. -2 The experiment involved UVB radiation and lasted for 6 days.

[0101] 1. Transepidermal water loss detection

[0102] Before the experiment, all groups of mice had their shaved areas photographed with a camera and dermoscopically examined. The TEWL value and skin moisture content of the same location were measured (three measurements were taken for the same area, and the average value was used). The results are shown in Table 5.

[0103] Table 5

[0104] As shown in Table 5, the average TWEL value of mice in the CTL group did not change significantly before and after the experiment, while the average TWEL value of the MOD group increased significantly after light exposure compared to before light exposure. The TWEL values ​​of the N1, N2, N3, and Y groups all increased significantly compared to before light exposure. However, compared with the MOD group, the increase in TWEL values ​​of the N1, N2, N3, and Y groups was significantly reduced. Among them, the TWEL values ​​of the N1 group after light exposure were similar to those of the Y group, which also indicates that the effect of the N1 group in repairing photodamaged skin is similar to that of chemical reagents.

[0105] Regarding changes in skin moisture content, the normal CTL group showed no significant change in skin moisture content before and after the experiment. In the MOD group, the skin moisture content of mice after light exposure was significantly lower than before, indicating that long-term ultraviolet radiation can lead to dry skin, reduced moisture content, and impaired skin barrier function. While the skin moisture content of groups N1, N2, N3, and Y was significantly lower than before light exposure, the decrease was significantly less pronounced compared to the MOD group, demonstrating that the gel preparation prepared in this invention has a significant effect on repairing skin barrier damage caused by photodamage.

[0106] 2. Skin condition score

[0107] The scoring criteria for this experiment are shown in Table 6. Figure 5 As shown in Table 1 (the corresponding appearance conditions of the mouse dorsal skin photodamage are given according to the 0~3.0 scores), scores were given according to the scoring criteria after the experiment.

[0108] Table 6

[0109] The scoring results are shown in Table 6. Table 6 shows that gel formulation 1 has the most significant repair effect on photodamage to the skin. A comparison of the results of groups N1 and N2 shows that the lyophilized protectant also plays a certain repair role in photodamage to the skin. This is because the increased glycerol content serves two purposes: firstly, glycerol protects the integrity of exosomes; secondly, it moisturizes and protects the skin barrier. Additionally, the increased ceramide content reduces inflammation and protects exosomes from aggregation and membrane fusion. Compared with the positive control group Y, group N's damage repair effect is even better than group Y. This clearly demonstrates that the protective and repairing power of the gel formulation of this invention against photodamage-induced skin barrier damage is superior to that of chemical formulations. Comparing groups N1 and N3, the damage repair effect of group N3 is significantly less than that of group N1. This is because during the lyophilized storage of exosomes, some exosomes experience a decrease in activity, leading to a significant reduction in the repair effect.

Claims

1. An exosome composition, characterized in that, The exosome composition consists of umbilical cord mesenchymal stem cell exosomes, honeysuckle exosomes, and green tea exosomes in a weight ratio of 2-3:0.5-0.8:1-2.

2. The use of the exosome composition of claim 1 in the development of products that alleviate and / or treat skin barrier damage.

3. The application according to claim 2, characterized in that, The exosome composition has at least one of the following application effects: (1) Promotes the proliferation and / or migration of immortalized human keratinocytes; (2) Promotes FLG expression in immortalized human keratinocytes; (3) Inhibits the expression of inflammatory factors in human immortalized keratinocytes.

4. The lyophilization protectant for the exosome composition of claim 1, characterized in that, The lyophilization protectant consists of 0.3-0.5% v / v ceramide, lysine, and 1.8-1.5% v / v glycerol, wherein the final concentration of lysine after mixing the exosome lyophilization protectant and exosomes is 70-80 mM.

5. The use of the exosome composition of claim 1 and the lyophilization protectant of claim 4 in the preparation of products that alleviate and / or treat skin barrier damage.

6. A gel formulation comprising the exosome composition of claim 1 and the lyophilization protectant of claim 4.

7. The use of the gel formulation of claim 6 in the preparation of products for relieving and / or treating skin barrier damage.

8. The application according to claim 7, characterized in that, The skin barrier damage mentioned refers to damage to the skin barrier caused by ultraviolet radiation.

9. The application according to claim 8, characterized in that, The skin barrier damage caused by ultraviolet radiation is manifested by an increase in TWEL value and a decrease in the moisture content of the stratum corneum.

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