A dendrobium exosome-containing skin care composition and a preparation method thereof

Abstract

The application discloses a skin care composition containing dendrobium exosome and various traditional Chinese medicine extracts and a preparation method thereof. The composition takes dendrobium exosome as a core active ingredient, combines with plant extracts such as dandelion, wild chrysanthemum and radix scutellariae, realizes the effects of anti-inflammation, anti-oxidation and promotion of skin repair through multi-component synergistic action, and can effectively improve acne-related inflammation and delay skin aging. On this basis, the application further constructs a delivery system taking gel as a carrier, improves the stability of dendrobium exosome and its retention and slow-release performance in the skin, and thus enhances the overall efficacy. The application also provides a preparation method of the composition, including the separation and purification of dendrobium exosome, the preparation of plant extracts and the compounding process of the composition. The composition can be prepared into various external preparations such as gels, emulsions, creams, serums and masks, and has a good application prospect.

Description

Technical Field

[0001] This invention relates to the field of cosmetics and topical skin preparations, specifically to a skin care composition containing exosome-like vesicles derived from Dendrobium and plant extracts, and its preparation method, particularly to a skin repair composition using a gel as a carrier. Background Technology

[0002] With the increasing demand for skin care, topical preparations with anti-inflammatory, antioxidant, and skin-repair-promoting functions have received widespread attention. However, most existing skin care products use single active ingredients or simple compound systems, which still have limitations in improving acne-related inflammation and delaying skin aging.

[0003] In recent years, exosomes, as natural nanoscale vesicles, have been increasingly applied in the field of skin care, as they can carry bioactive substances such as proteins, lipids, and nucleic acids and play an important role in intercellular signal transduction and tissue repair. However, the application of exosomes in topical formulations still faces challenges such as insufficient stability, easy degradation, and limited retention time on the skin surface, affecting their actual efficacy.

[0004] On the other hand, extracts of traditional Chinese medicine such as dandelion, wild chrysanthemum, and scutellaria baicalensis possess certain anti-inflammatory and antioxidant activities, but their effects are limited when used alone, and the synergistic effects between different components have not been fully explored. Furthermore, existing research on the compounding methods of these plant extracts and their combined application with exosomes remains relatively limited.

[0005] In addition, in terms of delivery systems, existing skin care formulations often struggle to balance the stability and release efficiency of active ingredients, especially for exosome-based active substances, where there is a lack of effective carrier systems to protect and regulate their release.

[0006] Therefore, developing a skincare composition that can improve exosome stability, enhance the synergistic effect of multiple components, and possess good delivery performance is of great research significance and application value. Summary of the Invention

[0007] Purpose of the invention: The purpose of this invention is to provide a skin care composition and its preparation method to solve the problems of poor stability and low utilization efficiency of exosomes in topical preparations, as well as insufficient synergistic effect of multiple plant extracts in the prior art, thereby improving the utilization effect of active ingredients on the skin surface.

[0008] Technical Solution: To achieve the above objectives, the present invention provides the following technical solution:

[0009] A skin care composition comprising exosome-like vesicles derived from Dendrobium and a plant extract system, wherein the plant extract system comprises one or more of dandelion, wild chrysanthemum, and scutellaria baicalensis, and may further comprise one or more of salvia miltiorrhiza, forsythia, poria cocos, and angelica dahurica.

[0010] In one embodiment, the composition is a gel formulation.

[0011] In a preferred embodiment, the composition is a hydrogel system.

[0012] In one embodiment, the hydrogel system may be constructed from one or more of sodium alginate, carbomer, or cellulose thickeners to create a stable hydration environment on the skin surface and facilitate the sustained release of the active ingredients.

[0013] In a preferred embodiment, the components in the composition are as follows by weight: 12 parts Dendrobium nobile, 15 parts Taraxacum mongolicum, 10 parts Chrysanthemum indicum, 10 parts Scutellaria baicalensis, 12 parts Salvia miltiorrhiza, 10 parts Forsythia suspensa, 5 parts Poria cocos, and 5 parts Angelica dahurica.

[0014] In a preferred embodiment, the Dendrobium exosome-like vesicles are prepared by the following method: Dendrobium tissue is homogenized, then subjected to low-speed centrifugation to remove impurities, followed by membrane filtration and ultrafiltration concentration, and further purified by density gradient centrifugation or dialysis to obtain Dendrobium exosome-like vesicles.

[0015] In a preferred embodiment, the plant extract is prepared by the following method: dandelion, wild chrysanthemum, scutellaria baicalensis, and other raw materials are extracted by water extraction or alcohol extraction, and the extract is concentrated to obtain the corresponding plant extract.

[0016] Traditional Chinese medicine believes that skin inflammation, acne, and premature skin aging are often related to pathogenesis such as "yin deficiency and dryness-heat," "excessive heat in the lungs and stomach," "damp-heat accumulation," and "heat toxin stagnation in the skin." Heat toxins and dampness accumulating in the skin can lead to redness, sensitivity, and impaired skin barrier function; over time, this depletes body fluids, causing the skin to lack nourishment, further resulting in dryness, roughness, oxidative damage, and skin aging. Therefore, improving skin problems should follow the principles of "clearing heat and detoxifying, nourishing yin and moisturizing dryness, anti-oxidative repair, and harmonizing the skin's microenvironment."

[0017] Based on the above theory, this invention selects exosome-like vesicles from Dendrobium officinale and combines them with plant extracts such as dandelion, scutellaria baicalensis, wild chrysanthemum, salvia miltiorrhiza, forsythia suspensa, poria cocos and angelica dahurica to form a skin care composition, and combines it with a gel carrier system to achieve comprehensive effects of anti-inflammatory, antioxidant, skin repair promotion and delaying skin aging.

[0018] Dendrobium, a traditional Chinese medicine belonging to the genus Dendrobium of the Orchidaceae family, possesses properties such as nourishing yin and moistening dryness, clearing heat and promoting body fluid production, and nourishing the stomach and benefiting yin. Modern research indicates that Dendrobium contains polysaccharides, polyphenols, and active small molecule components, exhibiting excellent antioxidant capabilities. It can increase skin hydration, enhance skin barrier function, and delay skin aging. This invention further utilizes Dendrobium-derived exosome-like vesicles as the core active ingredient. These exosomes have a nanoscale vesicle structure, capable of carrying active substances such as proteins, lipids, and nucleic acids, participating in intercellular information transmission, thereby regulating skin cell function, promoting the repair of damaged skin, and improving the skin microenvironment.

[0019] Dandelion extract is rich in polyphenols and flavonoids, possessing heat-clearing, detoxifying, anti-inflammatory, and antioxidant properties, which can help improve skin inflammation and promote skin repair. Scutellaria baicalensis extract, rich in flavonoids, can reduce micro-inflammation and oxidative damage in the skin, synergistically enhancing the overall antioxidant and soothing effects with dandelion extract. Wild chrysanthemum has wind-dispelling, heat-clearing, and anti-inflammatory properties, helping to reduce skin redness and irritation. Salvia miltiorrhiza, forsythia suspensa, Poria cocos, and Angelica dahurica, among other ingredients, respectively exert effects such as promoting blood circulation and repair, clearing heat and resolving lumps, strengthening the spleen and removing dampness, and reducing swelling and promoting tissue regeneration, collectively improving skin condition.

[0020] Furthermore, this invention utilizes a gel system as a carrier for active ingredients, particularly a hydrogel system, which can create a stable, highly hydrated environment, improving the stability and skin adhesion of active ingredients, and prolonging the release time of exosomes and plant active ingredients, thereby enhancing skin absorption efficiency and localized effects. Compared to traditional skincare systems, the gel carrier of this invention has certain advantages in terms of active ingredient protection, sustained-release performance, and skin feel.

[0021] The present invention also provides a method for preparing the skin care composition, comprising the following steps:

[0022] (1) Dendrobium tissue was homogenized, centrifuged, filtered and ultrafiltered or density gradient centrifuged to obtain Dendrobium exosome-like vesicles;

[0023] (2) Extract the plant raw materials such as dandelion, wild chrysanthemum, and scutellaria baicalensis with water or alcohol, and then concentrate them to obtain plant extracts;

[0024] (3) Add the plant extract to the carrier system and mix thoroughly;

[0025] (4) Add Dendrobium exosome-like vesicles under low-speed conditions, homogenize, and degas;

[0026] (5) Adjust the pH of the system to obtain the skin care composition.

[0027] Beneficial effects: Compared with the prior art, the present invention has the following advantages:

[0028] (1) The present invention introduces exosome-like vesicles derived from Dendrobium as a delivery carrier for active ingredients, which helps to improve the stability and utilization efficiency of active substances;

[0029] (2) By combining extracts of plants such as dandelion, wild chrysanthemum, and scutellaria, the anti-inflammatory and antioxidant effects are enhanced to a certain extent, achieving synergistic effects of multiple components;

[0030] (3) Using gel formulations, especially hydrogel systems, as carriers helps to form a stable hydration environment on the skin surface, thereby prolonging the release time of active ingredients and improving their local effects.

[0031] (4) The composition of the present invention has good comprehensive properties in anti-inflammatory, antioxidant and skin repair promotion, and has good application prospects.

[0032] (5) The preparation process of the composition of the present invention is simple, has good repeatability, and is easy to industrialize and promote. Attached Figure Description

[0033] Figure 1 Transmission electron microscopy image of Dendrobium exosomes

[0034] Figure 2 Flowchart for the preparation and functional verification of Dendrobium exosome-like vesicle skincare composition

[0035] Figure 3 Comparison of appearance of different gel systems: Left bottle: HPMC-Carbomer system gel; Right bottle: Hydrogel

[0036] Figure 4 Line graphs showing the changes of each sample group in the ABTS radical system

[0037] Figure 5 Line graphs showing the changes of each sample group in the DPPH radical system

[0038] Figure 6 Effects of different doses of Dendrobium exosome composition on procollagen PICP secretion

[0039] Figure 7 Effects of different treatment groups on the activities of MMP-1, MMP-2 and MMP-9 (in vitro)

[0040] Figure 8 Effects of different treatment groups on the expression of aging biomarkers

[0041] Figure 9 Effects of different doses of Dendrobium exosome composition on IL-6 inflammatory factor levels

[0042] Figure 10Effects of different doses of Dendrobium exosome composition on TNF-α inflammatory factor levels

[0043] Figure 11 Effects of different doses of Dendrobium exosome composition on IL-1β inflammatory factor levels

[0044] Figure 12 Effects of Dendrobium exosome composition on MMP activity (in vivo) Detailed Implementation Plan

[0045] To make the technical solution of the present invention clearer, the present invention will be further described below with reference to specific embodiments. Those skilled in the art should understand that various modifications or substitutions can be made to the embodiments without departing from the spirit and essence of the present invention, and such modifications or substitutions should all fall within the protection scope of the present invention.

[0046] Example 1: Preparation of Dendrobium exosome-like vesicles

[0047] Take 500g of fresh Dendrobium nobile, wash and chop it, add an equal volume of PBS buffer and homogenize for 5-10 min. Centrifuge the resulting homogenate at 3000 × g for 10 min at 4 ℃, take the supernatant and filter it through 0.45 μm and 0.22 μm filter membranes respectively.

[0048] The filtrate was concentrated to approximately 1 / 10 of its original volume using an ultrafiltration membrane with a molecular weight cutoff of 100 kDa, and then purified by density gradient centrifugation or dialysis to obtain Dendrobium exosome-like vesicles. The obtained product can be stored at 4 °C or further prepared into a lyophilized powder for later use.

[0049] Example 2: Preparation of plant extracts

[0050] The following ingredients were selected as raw materials: 625 g of dandelion, 417 g of wild chrysanthemum, 417 g of scutellaria baicalensis, 500 g of salvia miltiorrhiza, 417 g of forsythia suspensa, 208 g of poria cocos, and 208 g of angelica dahurica.

[0051] Dandelion and wild chrysanthemum were extracted with water or by reflux with 50% ethanol, twice for 2 hours each time, and the extracts were combined and concentrated. Scutellaria baicalensis and Salvia miltiorrhiza were extracted by reflux with 70% ethanol, and the contents of flavonoids and tanshinone were standardized after concentration. Forsythia suspensa, Poria cocos, and Angelica dahurica were extracted with water or by low-concentration ethanol and then concentrated. The final plant extracts were prepared for use.

[0052] Example 3: Construction of different formulation combinations

[0053] To evaluate the effect of different component combinations on the performance of the skincare composition, the raw materials were combined as follows:

[0054] Group A: Dendrobium nobile exosome-like vesicles; Group B: Dandelion extract; Group C: Chrysanthemum indicum extract; Group D: Compound plant extracts, including Scutellaria baicalensis, Salvia miltiorrhiza, Forsythia suspensa, Poria cocos, and Angelica dahurica.

[0055] Based on this, different combination systems were constructed, including A, AD, BCD, ABD, ACD and ABCD combinations, for subsequent evaluation of antioxidant performance.

[0056] Example 4: Validation of the antioxidant performance of different combination systems

[0057] To evaluate the antioxidant capacity of different combination systems, the DPPH free radical scavenging experiment was used for determination.

[0058] Take samples from each combination system, dilute them appropriately, and weigh or prepare solutions of a certain concentration. Add 10 μL of sample solution to a 96-well plate, add 190 μL of DPPH solution (0.1 mM, dissolved in anhydrous ethanol), react in the dark for 30 min, and then measure the absorbance at 517 nm.

[0059] The same method was used to perform ABTS free radical scavenging experiments to determine the antioxidant capacity of different combination systems.

[0060] The experimental results are shown in Tables 1 and 2.

[0061] Table 1. Absorbance and relative change of each sample combination in the ABTS radical system

[0062]

[0063] Note: Compared with the blank control group, *P < 0.05, **P < 0.01. Statistical analysis was performed using one-way ANOVA. The same applies to the following tables.

[0064] As shown in Table 1, different sample combinations all exhibited certain antioxidant activities in the ABTS radical system. Among them, the ABCD group had the strongest scavenging ability and was significantly better than the blank control group under all concentration conditions (P < 0.01), showing excellent antioxidant performance.

[0065] Table 2. Absorbance and variation of each sample combination in the DPPH radical system

[0066]

[0067] This experiment evaluated the antioxidant capacity of different combinations of samples using ABTS and DPPH free radical systems. The results showed that each combination possessed a certain free radical scavenging ability, which increased with increasing concentration, exhibiting a good concentration-dependent effect. The ABCD combination showed superior antioxidant performance in both systems, indicating a synergistic enhancement effect among the components rather than a simple additive effect, thus significantly improving the overall antioxidant performance.

[0068] Example 5: Verification of the anti-aging effects of different combination systems

[0069] Human-derived fibroblasts (HDFs) were selected as an in vitro model to evaluate the anti-aging effects of the compositions of the present invention.

[0070] After cell seeding and culture, different treatment groups were set up, including:

[0071] Group A (Dendrobium officinale-derived components), Group B (Dandelion), Group C (Chrysanthemum indicum), Group D (a compound of Scutellaria baicalensis, Salvia miltiorrhiza, Forsythia suspensa, Poria cocos, and Angelica dahurica), and their combination groups (such as ABCD, ABD, ACD, BCD, etc.). A positive control group (TGF-β1, 5 ng / mL) and a solvent control group were also set up.

[0072] Each sample was prepared as an equivalent dilution of the skincare composition system of the present invention, and added to the cell culture system at different concentrations (10 μg / mL, 50 μg / mL, 200 μg / mL) for 24–72 h. After treatment, the following indicators were detected: collagen synthesis level (including procollagen PICP and total collagen content), MMP-1, MMP-2 and MMP-9 activities, and the expression levels of aging-related markers p16 and p21. Cellular compatibility of the samples was evaluated using cell viability assays.

[0073] Experimental results show that, compared with the solvent control group, the composition of the present invention can significantly promote collagen synthesis in fibroblasts, inhibit MMPs activity and reduce collagen degradation, while downregulating the expression levels of p16 and p21. No obvious cytotoxicity was observed within the effective concentration range.

[0074] Further comparison of different combinations revealed that the ABCD combination group showed better performance in all of the above indicators, suggesting a synergistic effect among the components.

[0075] The above results demonstrate that the composition of the present invention has good potential in delaying skin aging and promoting skin repair, and that there is a synergistic enhancement effect among the components, rather than a simple additive effect.

[0076] Table 3. Effects of different treatment groups on procollagen PICP secretion

[0077]

[0078] Note: Compared with the solvent control group, *P < 0.05, **P < 0.01; statistical analysis was performed using one-way ANOVA. The same applies to the following tables.

[0079] PICP test results showed that all treatment groups could promote procollagen secretion to varying degrees, with the ABCD combination group being significantly higher than the solvent control group (P < 0.01), approaching the level of the positive control.

[0080] Table 4. Effects of different treatment groups on MMP activity

[0081]

[0082] The results of MMP activity assay showed that the ABCD combination group had significant inhibitory effects on MMP-1, MMP-2 and MMP-9 (P < 0.01), and its inhibitory effect was close to that of the positive control.

[0083] Table 5. Effects of different treatment groups on the expression of aging biomarkers

[0084]

[0085] The results of aging-related biomarker detection showed that the ABCD combination group could significantly downregulate the expression levels of p16 and p21 (P < 0.01), and its effect was close to that of the positive control, suggesting that it has a certain anti-aging effect.

[0086] Example 6: Preparation of the composition gel formulation

[0087] To improve the stability of active ingredients and their sustained-release properties on the skin, this study constructed HPMC-carbomer composite gel system and sodium alginate hydrogel system, and compared their gelation state, flowability and performance.

[0088] (1) HPMC-Carbomer composite system: Take 0.30 g HPMC and add it to 50 mL of water. After dispersing it thoroughly under stirring, let it stand overnight at 4 ℃ to allow it to fully hydrate. Take another 0.10 g of carbomer and add it to 10 mL of water to moisten it for later use. Add 20 mL of the Chinese herbal extract to the above HPMC solution and mix well. Then add the carbomer dispersion and continue stirring. Subsequently, add triethanolamine dropwise to adjust the pH of the system to about 6.5 so that the system forms a gel. Finally, add 5 mL of Dendrobium exosome vesicle suspension under low-speed stirring. After mixing well, add water to 100 mL to obtain HPMC-Carbomer composite gel.

[0089] (2) Sodium alginate hydrogel system: Take 1.5 g sodium alginate and add it to 55 mL of water. Stir until completely dissolved to form a homogeneous solution. Add 30 mL of Chinese herbal extract and mix well. Then add 5 mL of Dendrobium exosome vesicle suspension under low-speed stirring. Then slowly add 1-2 mL of CaCl2 solution to carry out cross-linking reaction to form a soft hydrogel system.

[0090] Comparison of the two gel systems revealed that the sodium alginate hydrogel system exhibited a more uniform appearance, better fluidity, and a simpler preparation process. It also demonstrated good stability and biocompatibility, which is more conducive to the release of active ingredients and skin absorption. Therefore, the sodium alginate hydrogel system was preferentially selected as the final formulation system for subsequent experiments.

[0091] Example 7: Verification of the in vivo anti-inflammatory effect of the composition gel

[0092] An ICR mouse model of skin inflammation was induced using Propionibacterium acnes. The experimental animals were randomly divided into a model group, an ABCD combination treatment group (different doses), and a positive control group.

[0093] After the model was established, the composition of the present invention was prepared into a hydrogel formulation and applied topically at a certain dose (0.1–0.5 g / animal) once a day to the inflamed site for 7–14 days.

[0094] After the experiment, serum and skin tissue samples were collected to detect the levels of inflammatory factors IL-6, TNF-α and IL-1β, and the activities of MMP-1, MMP-2 and MMP-9 were measured. Skin histology was also observed.

[0095] The results showed that, compared with the model group, the composition of the present invention could significantly reduce the level of inflammatory factors and inhibit the activity of MMPs, and showed a certain dose-dependent trend; the inflammatory response of skin tissue was reduced and the structure was significantly restored.

[0096] Table 6. Effects of Dendrobium exosome composition on inflammatory factor levels

[0097]

[0098] Note: Compared with the model group, *P < 0.05, **P < 0.01. The same applies to the following table.

[0099] As shown in Table 6, the Dendrobium exosome composition can reduce the activity of MMP-1, MMP-2 and MMP-9 to varying degrees, and exhibits a certain dose-dependent effect. Compared with the model group, the medium and high dose groups showed significant differences (P < 0.01), with the high dose group showing an inhibitory effect close to that of the positive control group.

[0100] Table 7. Effects of Dendrobium exosome composition on MMP activity

[0101]

[0102] As shown in Table 7, the Dendrobium exosome composition significantly reduced the levels of inflammatory factors IL-6, TNF-α, and IL-1β, with a marked decreasing trend with increasing dosage. Compared with the model group, the medium- and high-dose groups showed significant differences (P < 0.01), with the high-dose group exhibiting effects close to the positive control group, suggesting good anti-inflammatory activity.

[0103] This invention has good application prospects and industrialization value.

Claims

1. A skincare composition, characterized in that, The composition comprises exosome-like vesicles derived from Dendrobium and a plant extract system, wherein the plant extract system comprises one or more of dandelion, wild chrysanthemum, and scutellaria baicalensis, and may further comprise one or more of salvia miltiorrhiza, forsythia suspensa, poria cocos, and angelica dahurica; the exosome-like vesicles are used as delivery carriers for the active ingredients to participate in the functional realization of the composition.

2. The skincare composition according to claim 1, characterized in that: The composition is a gel formulation.

3. The skincare composition according to claim 2, characterized in that: The gel is a hydrogel system.

4. The skincare composition according to claim 3, characterized in that: The hydrogel system includes one or more of sodium alginate, carbomer, or cellulose-based thickeners.

5. The skincare composition according to claim 1, characterized in that: The components in the composition are as follows by mass: Dendrobium nobile (used for preparing exosomes) 12 parts, dandelion 15 parts, wild chrysanthemum 10 parts, Scutellaria baicalensis 10 parts, Salvia miltiorrhiza 12 parts, Forsythia suspensa 10 parts, Poria cocos 5 parts, and Angelica dahurica 5 parts.

6. The skincare composition according to claim 5, characterized in that: The dosage of each component is allowed to be adjusted by ±20% based on the formula.

7. The skincare composition according to claim 1, characterized in that: The composition can also be prepared as an emulsion, cream, serum or mask.

8. A method for preparing the skincare composition as described in claim 1, characterized in that, include: Dendrobium exosomes were obtained by homogenizing, centrifuging, filtering, and ultrafiltration or density gradient centrifugation of Dendrobium tissue. Extracts are obtained by water extraction or alcohol extraction of dandelion, wild chrysanthemum, scutellaria baicalensis and other plant materials; the extracts are mixed with a gel matrix or a pharmaceutically acceptable carrier, and dendrobium exosomes are added. After homogenization, degassing and pH adjustment, the skin care composition is obtained.

9. The use of the skin care composition according to claim 1 in the preparation of topical anti-inflammatory or acne-improving preparations.

10. The use of the skincare composition according to claim 1 in the preparation of topical formulations for anti-oxidation, skin repair, or delaying skin aging.

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