Skin care composition, preparation method and application

Abstract

The invention belongs to the technical field of cosmetics, and particularly relates to a skin care composition, a preparation method and application, the skin care composition is composed of temperature-sensitive hydrogel, a 3D exosome and a traditional Chinese medicine extract, and the 3D exosome is prepared by culturing on the basis of a 2D exosome. According to the technical scheme, the temperature-sensitive hydrogel, the 3D exosome and the traditional Chinese medicine extract are used to replace cosmetic substrates, by taking a mask as an example, the skin-care composition can be coated on the face in a mode of using a moisturizer, a gel film layer is formed after the skin-care composition is in contact with the skin of the face, and the skin-care composition can be directly permeated and absorbed after being used, so that the use convenience is improved. According to the technical scheme, the skin care composition shows a 3D structure and is rich in fine gaps after being gelled, medicine components are contained in the gaps, and when the skin care composition is used, medicine borne by the skin care composition is released durably and stably.

Description

Technical Field

[0001] The present invention belongs to the technical field of cosmetics, and in particular relates to a skin care composition, a preparation method and an application thereof. Background Art

[0002] The skin is the most obvious external sign of aging. As a complex organ, it extends throughout the entire body. With aging, the skin's metabolic cycle gradually lengthens. Starting at age 25, as skin metabolism slows, the elastic fibers in the dermis begin to break and deform. Aged cells continue to form and remain in the epidermis for extended periods, unable to be replaced by new cells. This leads to a continuous decrease in skin elasticity and moisture, resulting in sagging skin, wrinkles, and a loss of radiance. As living standards improve, people's pursuit of skin care is becoming increasingly demanding. Besides cosmetic surgery, most people prefer to use high-quality skincare products to gently maintain their youthful appearance.

[0003] On the other hand, facial acne, dark spots, and other issues that affect appearance have received significant attention. Existing technologies suggest that mesenchymal stem cells have multidirectional differentiation capabilities and can aid in the repair of damaged tissues, leading to their increasing popularity in skincare products. However, existing skincare products containing mesenchymal stem cells do not actually deliver the expected results. Summary of the Invention

[0004] The object of the present invention is to provide a skin care composition, a preparation method and an application thereof, so as to solve the problems existing in the above-mentioned background technology.

[0005] To achieve the above objectives, this application is implemented through the following technical solutions:

[0006] A skin care composition consists of a thermosensitive hydrogel, 3D exosomes and a traditional Chinese medicine extract. The 3D exosomes are prepared by using 2D cells through a 3D culture system.

[0007] Furthermore, the temperature-sensitive hydrogel is one or a combination of α-ketoglutaric acid, carboxyl chitin, hydroxypropyl chitin, hydroxybutyl chitosan, and hydroxypentyl chitosan.

[0008] Furthermore, the skin care composition is composed of 10-25% by weight of 3D exosomes, 10-15% by weight of a traditional Chinese medicine extract, and the remainder by weight of a thermosensitive hydrogel.

[0009] Furthermore, the Chinese herbal medicine extract includes one or more of Angelica dahurica extract, Aloe vera extract or Bombyx batryticatus extract.

[0010] Furthermore, the angelica dahurica extract and the aloe vera extract are extracts obtained by sequentially extracting with water and then with n-hexane, and the silkworm blast extract is an extract obtained by first extracting with ethanol and then with chloroform.

[0011] Furthermore, when there are two or more Chinese herbal extracts, they are used in equal amounts.

[0012] A method for preparing any of the above skin care compositions comprises the following steps:

[0013] S1. Enrich 3D exosomes using 2D cells through a 3D culture system;

[0014] S2, preparing Chinese herbal medicine extracts;

[0015] S3. Mix the thermosensitive hydrogel, 3D exosomes and traditional Chinese medicine extract according to a set ratio to obtain a skin care composition.

[0016] An application, use of any of the above skin care combinations in cosmetics.

[0017] Further, the application in facial masks.

[0018] The beneficial effects of the present invention are:

[0019] The skin care composition of the present technical solution replaces the use of cosmetic bases by using thermosensitive hydrogels, 3D exosomes and traditional Chinese medicine extracts. Taking facial masks as an example, they can be applied to the face in the same way as moisturizers. After contacting the facial skin, a gel film layer is formed. After use, it can be directly penetrated and absorbed, thereby improving the convenience of use.

[0020] The skin care composition of the present technical solution presents a 3D structure after the product gels and is rich in tiny voids, in which the drug components are contained. When used, the drugs carried by the product are released in a long-lasting and stable manner. DETAILED DESCRIPTION

[0021] The technical solutions of the present invention are described in detail below through examples. The following examples are merely exemplary and can only be used to explain and illustrate the technical solutions of the present invention, and cannot be interpreted as limiting the technical solutions of the present invention.

[0022] The present application provides a skin care composition. The specific type of skin care product is not specified in the present application, and those skilled in the art can determine the specific type of skin care product as needed.

[0023] The skin care composition of the present application consists of three components: thermosensitive hydrogel, 3D exosomes and Chinese medicine extracts, wherein the 3D exosomes are prepared by 2D cells through a 3D culture system.

[0024] In the present application, the thermosensitive hydrogel is one or a combination of more than one of α-ketoglutaric acid, carboxyl chitin, hydroxypropyl chitin, hydroxybutyl chitosan, and hydroxypentyl chitosan. The thermosensitive hydrogel of the present application has thermosensitivity and water solubility. At low temperatures (generally referring to temperatures below the surface temperature of human skin), the product is a liquid with fluidity. Under the influence of human body temperature, it quickly transforms into a non-flowing gel. The present application utilizes the fluidity of the product at low temperatures to completely cover complex wounds such as gap-type and wrinkled wounds. Under the influence of body temperature, it quickly transforms into a gel, tightly adhering to the wound surface without loss. Moreover, after gelation, the product exhibits a 3D structure rich in tiny pores that can accommodate drug molecules. The drug carried by the product is released in a sustained and stable manner, acting directly on the wound surface, solving the problems of severe loss and short duration of drug efficacy in clinical local application. It provides an ideal drug delivery model with the highest efficacy and the least side effects in clinical practice.

[0025] The thermosensitive hydrogels mentioned above in this application are all recognized to have good biosafety, and the clinical advantages of hemostasis, healing promotion, antibacterial and scar inhibition.

[0026] The skin care composition of the present application also uses Chinese herbal extracts, including one or more of Angelica dahurica extract, Aloe vera extract, or Bombyx batryticatus extract. The Chinese herbal extracts involved in the present application are all secondary extracts, rather than primary extracts, which is the key to the present application. Among them, the specific preparation method of the Angelica dahurica and Aloe vera extract is:

[0027] The Chinese medicinal raw material Angelica dahurica is first boiled twice with 4-6 times the amount of water according to the conventional boiling method of Chinese medicine, and the medicinal liquids after the two boilings are combined for concentration. The specific concentration also adopts the existing conventional method, such as conventional concentration or reduced pressure concentration. After the medicinal liquid is concentrated to 1 times, n-hexane of the same volume as the medicinal liquid is added, stirred and then allowed to stand for extraction for 4-12 hours. After removing the n-hexane from the n-hexane part, the obtained product is the Angelica dahurica extract.

[0028] Crush fresh aloe vera and add 4-6 times water to boil once, then filter to remove the aloe residue to obtain aloe vera liquid, then concentrate the aloe vera liquid by 1 time, or further concentrate as needed to obtain a concentrated solution, then add n-hexane of the same volume as the concentrated solution to the concentrated solution, stir, and then stand for extraction for 4-12 hours. After removing the n-hexane from the n-hexane portion, the obtained product is aloe vera extract.

[0029] Bombyx batryticatus was extracted with anhydrous ethanol, 85% ethanol and 70% ethanol in sequence. The extracts were combined and then deethanolized to obtain Bombyx batryticatus ethanol extract. The Bombyx batryticatus ethanol extract was then dissolved in 4-8 times of chloroform. After standing for 8-12 hours, the insoluble matter was removed and then dechloroformed to obtain Bombyx batryticatus extract.

[0030] In the present application, if the Chinese herbal medicine extract is not a single component, the dosage ratio of each Chinese herbal medicine extract is usually equal.

[0031] In this application, the 3D exosomes of this application are prepared by culturing based on 2D exosomes, and the culturing process includes the following steps:

[0032] This culture process uses Huaxin Bio 3D FloTrix ® The vivaEXO exosome bioreactor has the following specific steps:

[0033] The bioreactor was wiped clean with 75% lees and placed in a CO2 incubator (5% CO2, 37°C) and connected to the power supply of the bioreactor.

[0034] Move four sterile 500 mL spinner flasks to the laminar flow hood.

[0035] The 2D cultured cells were photographed to confirm that the cell morphology was long spindle-shaped and the confluence was above 80%. The 2D cultured cells were digested, and after digestion, the cells were counted and the cell viability was detected.

[0036] Add 800 mg of microcarriers to each spinner flask, then add 50 mL of 3D cell culture medium to each spinner flask and gently shake the spinner flask to disperse the microcarriers.

[0037] After all the microslides were evenly suspended in the culture medium, 2.0×10 7 cells (i.e. 5×10 5 cells / 20mg microslide).

[0038] Each spinner flask was supplemented with culture medium to 150 mL, and then placed on the bioreactor and set to a rotation speed of 30 rpm for 5 min. The spinner flask was then left to stand for 1 h as one cycle, and a total of 24 cycles were performed.

[0039] 3D culture system observation (first day):

[0040] Move the spinner flask to the 3D FloTrix in the laminar flow hood. ® On the miniSpin bioreactor, the rotation speed was set to 40 rpm. After the microtissue was evenly suspended in the entire culture system, an electric pipette was used to sample from the side arm.

[0041] Take 10 μL of cell culture supernatant, add it dropwise to the glucose test strip, and read the glucose content.

[0042] 100 μL of microtissue suspension was placed in a 96-well plate, the culture supernatant was discarded, 100 μL of fluorescent staining working solution for living cells was added, and the plate was incubated at room temperature in the dark for 30 min; the staining solution was discarded and 100 μL of PBS was added, the plate was observed under a fluorescence microscope, and photographed.

[0043] Take three 1 mL microtissue suspensions, discard 900 μL of supernatant, add 900 μL of 1 mg / mL lysis buffer, and place in a 37°C water bath for lysis for 30 min; mix the cell suspension, mix the cell suspension with 0.4% trypan blue dye in a 1:1 ratio, take 20 μL, add it to a cell counting plate, and count the cells.

[0044] Then, 350 mL of 3D cell culture medium was added to each spinner flask, and the rotation speed was adjusted to a constant speed of 40 rpm.

[0045] 3D culture system observation (day 4):

[0046] The steps of sample connection, glucose measurement, live cell fluorescence staining, microcarrier lysis and counting, and rehydration were the same as before.

[0047] 3D culture system observation (day 5):

[0048] The steps of sample connection, glucose measurement, live cell fluorescence staining, and microcarrier lysis and counting were the same as before. 3L of cell culture supernatant was harvested and passed through Huakan Bio's 3D FloTrix ® The vivaEXO exosome harvesting device obtains 3D exosomes.

[0049] The present application provides a method for preparing the above-mentioned skin care composition, comprising the following steps:

[0050] S1. Prepare 3D exosomes from 2D cells using a 3D culture system;

[0051] S2, preparing Chinese herbal medicine extracts;

[0052] S3. Mix the thermosensitive hydrogel, 3D exosomes and traditional Chinese medicine extract according to a set ratio to obtain a skin care composition.

[0053] Example 1

[0054] S1. Prepare 3D exosomes from 2D cells using a 3D culture system;

[0055] S2, preparing Chinese herbal medicine extracts;

[0056] S3. Mix 10% of 3D exosomes, 10% of Angelica dahurica extract, and the remainder of hydroxybutyl chitosan in proportion to obtain a skin care composition.

[0057] Example 2

[0058] S1. Prepare 3D exosomes from 2D cells using a 3D culture system;

[0059] S2, preparing Chinese herbal medicine extracts;

[0060] S3. Mix 25% of 3D exosomes, 7% of Angelica dahurica extract, 7% of Bombyx batryticatus extract and the remainder of hydroxybutyl chitosan in proportion to obtain a skin care composition.

[0061] Example 3

[0062] S1. Prepare 3D exosomes from 2D cells using a 3D culture system;

[0063] S2, preparing Chinese herbal medicine extracts;

[0064] S3. Mix 15% of 3D exosomes, 5% of Angelica dahurica extract, 5% of Bombyx batryticatus extract, 5% of aloe vera extract and the remainder of hydroxybutyl chitosan in proportion to obtain a skin care composition.

[0065] Example 4

[0066] S1. Prepare 3D exosomes from 2D cells using a 3D culture system;

[0067] S2, preparing Chinese herbal medicine extracts;

[0068] S3. Mix 20% of 3D exosomes, 15% of Bombyx batryticatus extract, and the remaining hydroxybutyl chitosan and hydroxypentyl chitosan in proportion, wherein the weight ratio of hydroxybutyl chitosan to hydroxypentyl chitosan is 3:1, to obtain a skin care composition.

[0069] Example 5

[0070] S1. Prepare 3D exosomes from 2D cells using a 3D culture system;

[0071] S2, preparing Chinese herbal medicine extracts;

[0072] S3. Mix 22% of 3D exosomes, 12% of aloe vera extract, and the remaining amounts of hydroxybutyl chitosan and hydroxypentyl chitosan in proportion, wherein the weight ratio of hydroxybutyl chitosan to hydroxypentyl chitosan is 5:1, to obtain a skin care composition.

[0073] The skin care compositions of the above embodiments are used in cosmetics, such as facial masks.

[0074] The above are only preferred embodiments of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principles of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A skin care composition, characterized in that It consists of thermosensitive hydrogel, 3D exosomes and Chinese medicine extracts. The 3D exosomes are prepared by 2D cells through a 3D culture system.

2. The skin care composition according to claim 1, characterized in that The temperature-sensitive hydrogel is one or a combination of α-ketoglutaric acid, carboxyl chitin, hydroxypropyl chitin, hydroxybutyl chitosan, and hydroxypentyl chitosan.

3. The skin care composition according to claim 1, characterized in that The skin care composition is composed of 10-25% by weight of 3D exosomes, 10-15% of traditional Chinese medicine extracts and the remainder of thermosensitive hydrogel.

4. The skin care composition according to claim 1, characterized in that The Chinese medicine extract comprises one or more of angelica dahurica extract, aloe vera extract or silkworm blast extract.

5. The skin care composition according to claim 4, characterized in that The angelica dahurica extract and the aloe vera extract are both extracts obtained by sequentially extracting with water and then with n-hexane, while the silkworm blastoderm extract is an extract obtained by first extracting with ethanol and then with chloroform.

6. The skin care composition according to claim 4, characterized in that When there are two or more Chinese herbal extracts, they should be used in equal amounts.

7. A method for preparing the skin care composition according to any one of claims 1 to 6, characterized in that: The following steps are involved: S1. Enrich 3D exosomes using 2D cells through a 3D culture system; S2, preparing Chinese herbal medicine extracts; S3. Mix the thermosensitive hydrogel, 3D exosomes and traditional Chinese medicine extract according to a set ratio to obtain a skin care composition.

8. An application, characterized in that: Use of the skin care combination according to any one of claims 1 to 7 in cosmetics.

9. The use according to claim 8, characterized in that Application in facial masks.