CN113368030A - Stem cell-derived exosome composition for epidermis repair, preparation method and application - Google Patents
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Abstract
The invention provides a stem cell-derived exosome composition for epidermis repair, a preparation method and application, and relates to the technical field of biological materials. The stem cell-derived exosome composition for epidermal repair comprises human adipose-derived exosomes. The invention also provides a preparation method of the stem cell-derived exosome composition for epidermis repair and application of the stem cell-derived exosome composition in skin care products and/or cosmetics. The stem cell-derived exosome composition for epidermis repair has the advantages of effectively repairing the epidermis of the skin and inhibiting the recurrence of epidermis damage problems such as allergy, redness, dry itching, acne and stabbing pain.
Description
Technical Field
The invention relates to the technical field of biological materials, in particular to a stem cell-derived exosome composition for epidermis repair, a preparation method and application.
Background
The skin of a person is composed of epidermis, dermis and hypodermis, and contains accessory organs (sweat glands, sebaceous glands, nails and toenails) as well as blood vessels, lymphatic vessels, nerves, muscles and the like. The epidermis is the outermost layer of the skin and the first barrier of the human body, resists various external harmful factors, has an average thickness of 0.2 mm, and can be divided into 5 layers from outside to inside according to different development stages and morphological characteristics of cells, wherein the 5 layers comprise a stratum corneum, a stratum lucidum, a stratum granulosum, a spinous cell layer and a basal layer.
With the continuous development of economy and the continuous change of environment, the skin of people is also easily affected by various aspects, so that various problems occur, such as allergy, redness, dryness and itching, acne, stabbing pain and the like, any skin care product cannot be used, and the life and the work of people are seriously affected. However, no skin preparation is available to deal well with the recurring skin problems of allergy, redness, acne, dry itching and stinging. The existing skin preparations, such as various skin care products and the like, mostly stay in the stratum corneum, have poor repairing effect on the skin epidermis with problems, and cannot meet the increasingly urgent skin requirements of people.
Thus, there is a need for a skin formulation that overcomes the aforementioned drawbacks.
Disclosure of Invention
The invention aims to provide a stem cell-derived exosome composition for epidermis repair, which can effectively repair the epidermis of skin and inhibit the recurrence of epidermis damage problems such as allergy, redness, dry itching, acne and stabbing pain.
The invention also aims to provide a preparation method of the stem cell-derived exosome composition for epidermis repair, which prepares raw materials by classification and combining advanced technologies such as low-temperature freeze drying and the like, and finally prepares the composition by using a simple process, so that the effect of repairing the epidermis by the composition is better.
The invention further aims to provide an application of the stem cell-derived exosome composition for epidermis repair in preparation of skin care products and/or cosmetics.
The technical problem to be solved by the invention is realized by adopting the following technical scheme.
In one aspect, the present embodiments provide a stem cell-derived exosome composition for epidermal repair, comprising human adipose stem cell-derived exosomes.
In another aspect, the present embodiments provide a method for preparing a stem cell-derived exosome composition for epidermal repair, comprising the steps of: respectively dissolving the rehmannia root extract, the ligusticum wallichii extract and the radix sileris extract in partial solvents, mixing to obtain a mixed solution I, adding the mixed solution I and the functional auxiliary agent into the residual solvent, mixing to obtain a mixture II, adding the composite gel freeze-dried powder, the human adipose mesenchymal stem cell freeze-dried powder and the human adipose stem cell-derived exosome at the temperature of 4 ℃, and homogenizing to obtain the stem cell-derived exosome composition for epidermis repair.
In a further aspect, the present application provides a use of the aforementioned stem cell-derived exosome composition for epidermal repair in the preparation of a skin care product and/or a cosmetic product.
In summary, compared with the prior art, the embodiments of the present invention have at least the following advantages or beneficial effects:
the stem cell-derived exosome composition for epidermis repair provided by the invention can effectively repair the epidermis of the skin and inhibit the recurrence of the problems of epidermis damage such as allergy, redness, dry itching, acne and stabbing pain.
In addition, the preparation method of the stem cell-derived exosome composition for epidermis repair provided by the invention prepares raw materials by classification and combining with advanced technologies such as low-temperature freeze drying and finally prepares the composition by using a simple process, so that the effect of repairing epidermis by the composition is better.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. The examples, in which specific conditions are not specified, were conducted under conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products available commercially.
In one aspect, the present embodiments provide a stem cell-derived exosome composition for epidermal repair, comprising human adipose stem cell-derived exosomes.
In some embodiments of the present invention, the exosome is prepared by culturing the collected adipose-derived mesenchymal stem cells normally until the cell fusion degree reaches 70-80%, replacing the culture medium, continuously culturing for 10-14 h, collecting the supernatant, and extracting with an exosome extraction kit to obtain the human adipose-derived exosome.
In some embodiments of the present invention, the stem cell-derived exosome composition for epidermal repair further comprises a composite gel lyophilized powder, wherein the composite gel is prepared by crosslinking and lyophilizing sodium hyaluronate and collagen.
In some embodiments of the present invention, the above stem cell-derived exosome composition for epidermal repair is prepared by preparing collagen into a collagen solution, preparing sodium hyaluronate into a sodium hyaluronate solution, mixing the collagen solution and the sodium hyaluronate solution, adding genipin into the mixture, crosslinking the mixture at 45 ℃ for 2.5 hours to obtain a mixture, pre-freezing the mixture, and freeze-drying the mixture at-80 ℃ to obtain the composite gel freeze-dried powder.
In some embodiments of the present invention, in the above stem cell-derived exosome composition for epidermal repair, the concentration of the collagen solution is 1.2 wt%, the concentration of the sodium hyaluronate solution is 1.0 wt%, the volume ratio of the collagen solution to the sodium hyaluronate solution is 1:2, and the final concentration of genipin in the mixture is 0.02 mol/L.
In some embodiments of the invention, the stem cell-derived exosome composition for epidermal repair described above, is lyophilized for 48 h.
In some embodiments of the present invention, the stem cell-derived exosome composition for epidermal repair described above further comprises a rehmannia glutinosa extract, a ligusticum wallichii extract and a saposhnikovia divaricata root extract.
In some embodiments of the invention, the stem cell-derived exosome composition for epidermis repair comprises, by weight, 0.8-1.2% of composite gel lyophilized powder, 1.0-4.0% of rehmannia root extract, 0.5-2.0% of ligusticum wallichii extract, 0.5-2.0% of saposhnikovia divaricata root extract, 0.5-0.8% of human adipose-derived exosome, and the balance of a solvent and a functional assistant.
In another aspect, the present embodiments provide a method for preparing a stem cell-derived exosome composition for epidermal repair, comprising the steps of: respectively dissolving the rehmannia root extract, the ligusticum wallichii extract and the radix sileris extract in partial solvents, mixing to obtain a mixed solution I, adding the mixed solution I and the functional auxiliary agent into the residual solvent, mixing to obtain a mixture II, adding the composite gel freeze-dried powder, the human adipose mesenchymal stem cell freeze-dried powder and the human adipose stem cell-derived exosome at the temperature of 4 ℃, and homogenizing to obtain the stem cell-derived exosome composition for epidermis repair.
In a further aspect, the present application provides a use of the aforementioned stem cell-derived exosome composition for epidermal repair in the preparation of a skin care product and/or a cosmetic product.
The features and properties of the present invention are described in further detail below with reference to examples.
Example 1
The present embodiment aims to provide a human adipose-derived exosome.
1. Culturing human adipose-derived mesenchymal stem cells
Collecting 10mL of fat of a normal person, cleaning with PBS for three times to remove stains, cutting, digesting with 0.12% type I collagenase at 37 ℃ for 30min, adding 2mL of 10% fetal calf serum to stop digestion after digestion, centrifuging at 1500g for 10min to obtain a precipitate, repeatedly sieving the precipitate with a 200-mesh cell sieve for 2 times after resuspension, collecting filtrate, centrifuging to obtain a precipitate, and resuspending the precipitate to 5 × 104Perml, inoculated in 6-well plates, placed at 37 ℃ in 5% CO2Culturing in an incubator, performing first liquid change after the cells are completely attached to the wall, changing the complete culture medium every 2 days after the first liquid change, adding 1ml of digestive juice (0.25% Trypsin-0.53mM EDTA) for digestion when the cells grow to 80% fusion, and performing passage at a ratio of 1:3 after digestion. 90% of DMEM high-sugar culture medium; 10% of fetal bovine serum.
2. Detection of human adipose-derived mesenchymal stem cells
The human adipose-derived mesenchymal stem cells (P3) were prepared into a cell suspension, and murine anti-human CD29, CD44, CD105, CD31 and CD45 monoclonal antibodies were added thereto. Incubating at 4 ℃ in the dark for 30min, washing with PBS for three times, adding PBS for resuspension, and detecting the cell surface antigen in an up-flow manner.
Flow detection shows that the CD29+、CD44+、CD105+、CD31-And CD45-As can be seen, the human adipose mesenchymal stem cell (P3) is a human adipose mesenchymal stem cell.
3. Collecting exosomes
And (3) carrying out passage on the human adipose-derived mesenchymal stem cell (P3) according to a ratio of 1:3 to obtain a human adipose-derived mesenchymal stem cell (P4), normally culturing the human adipose-derived mesenchymal stem cell until the cell fusion degree reaches 70-80%, replacing a serum-free culture medium, continuously culturing for 10-14 h, and collecting a supernatant. The supernatant was centrifuged at 1000 Xg for 20min at room temperature, and after removing cell debris, the supernatant was transferred to a 2ml centrifuge tube. And (4) operating according to the specification of the Invitrogen secretion separation kit, collecting the exosomes, and detecting the purity of the exosomes until the purity reaches over 95 percent and the protein concentration is 2.0 g/L.
4. Exosome detection
And (3) detecting the human adipose-derived mesenchymal stem cell exosomes by using an electron microscope, and displaying that the diameter of the exosomes is 60-100 nm.
Meanwhile, the human adipose-derived mesenchymal stem cell exosome is subjected to flow identification, and the detection result shows that the CD9 is CD9+、CD29+、CD44+And CD90+，CD31-And CD45-. Therefore, the exosome prepared in the embodiment is an exosome derived from human adipose mesenchymal stem cells.
Example 2
The present embodiment aims to provide a stem cell-derived exosome composition for epidermal repair, which is mainly prepared by the following steps:
1. preparation of composite gel
Preparing collagen into a 1.2 wt% collagen solution, preparing sodium hyaluronate into a 1.0% sodium hyaluronate solution, mixing the collagen solution and the sodium hyaluronate solution, adding genipin with a final concentration of 0.02mol/L into the mixture, crosslinking the mixture at 45 ℃ for 2.5 hours to obtain a mixture, pre-freezing the mixture, and then freeze-drying the mixture at-80 ℃ for 48 hours to obtain the composite gel freeze-dried powder.
2. Preparation of exosomes
Exosomes were prepared according to the procedure in example 1, with exosomes having a purity of over 96% and a concentration of 2.0g/L being selected.
3. Preparation of plant extract working solution
Respectively dissolving rehmanniae radix extract, rhizoma Ligustici Chuanxiong extract and radix Saposhnikoviae extract in water to obtain rehmanniae radix working solution, rhizoma Ligustici Chuanxiong working solution and radix Saposhnikoviae working solution, and mixing the rehmanniae radix working solution, rhizoma Ligustici Chuanxiong working solution and radix Saposhnikoviae working solution to obtain mixed solution I.
4. Mixing
Adding the mixed solution I and the functional auxiliary agent into the residual solvent, mixing to obtain a mixture II, adding the composite gel freeze-dried powder, the human adipose-derived stem cell freeze-dried powder and the human adipose-derived exosomes into the mixture II at 4 ℃, and homogenizing to obtain the stem cell-derived exosome composition for epidermis repair, so that the stem cell-derived exosome composition for epidermis repair finally comprises the following raw materials: 0.8 wt% of composite gel freeze-dried powder, 4 wt% of rehmannia root extract, 0.5 wt% of ligusticum wallichii extract, 0.5 wt% of radix sileris extract, 0.8 wt% of exosome derived from human adipose-derived stem cells, and the balance of solvent and functional additive, wherein the functional additive is cosolvent and is proper in amount.
Example 3
The present embodiment aims to provide a stem cell-derived exosome composition for epidermal repair, which is mainly prepared by the following steps:
1. preparation of composite gel
Preparing collagen into a 1.2 wt% collagen solution, preparing sodium hyaluronate into a 1.0% sodium hyaluronate solution, mixing the collagen solution and the sodium hyaluronate solution, adding genipin with a final concentration of 0.02mol/L into the mixture, crosslinking the mixture at 45 ℃ for 2.5 hours to obtain a mixture, pre-freezing the mixture, and then freeze-drying the mixture at-80 ℃ for 48 hours to obtain the composite gel freeze-dried powder.
2. Preparation of exosomes
Exosomes were prepared according to the procedure in example 1, with exosomes having a purity of over 96% and a concentration of 2.0g/L being selected.
3. Preparation of plant extract working solution
Respectively dissolving rehmanniae radix extract, rhizoma Ligustici Chuanxiong extract and radix Saposhnikoviae extract in water to obtain rehmanniae radix working solution, rhizoma Ligustici Chuanxiong working solution and radix Saposhnikoviae working solution, and mixing the rehmanniae radix working solution, rhizoma Ligustici Chuanxiong working solution and radix Saposhnikoviae working solution to obtain mixed solution I.
4. Mixing
Adding the mixed solution I and the functional auxiliary agent into the residual solvent, mixing to obtain a mixture II, adding the composite gel freeze-dried powder, the human adipose-derived stem cell freeze-dried powder and the human adipose-derived exosomes into the mixture II at 4 ℃, and homogenizing to obtain the stem cell-derived exosome composition for epidermis repair, so that the stem cell-derived exosome composition for epidermis repair finally comprises the following raw materials: 1.2 wt% of composite gel freeze-dried powder, 3.5 wt% of rehmannia root extract, 0.75 wt% of ligusticum wallichii extract, 0.75 wt% of radix sileris extract, 0.8 wt% of exosome derived from human adipose-derived stem cells, the balance of solvent and functional additive, wherein the functional additive is cosolvent and a proper amount of the cosolvent.
Example 4
The present embodiment aims to provide a stem cell-derived exosome composition for epidermal repair, which is mainly prepared by the following steps:
1. preparation of composite gel
Preparing collagen into a 1.2 wt% collagen solution, preparing sodium hyaluronate into a 1.0% sodium hyaluronate solution, mixing the collagen solution and the sodium hyaluronate solution, adding genipin with a final concentration of 0.02mol/L into the mixture, crosslinking the mixture at 45 ℃ for 2.5 hours to obtain a mixture, pre-freezing the mixture, and then freeze-drying the mixture at-80 ℃ for 48 hours to obtain the composite gel freeze-dried powder.
2. Preparation of exosomes
Exosomes were prepared according to the procedure in example 1, with exosomes having a purity of over 96% and a concentration of 2.0g/L being selected.
3. Preparation of plant extract working solution
Respectively dissolving rehmanniae radix extract, rhizoma Ligustici Chuanxiong extract and radix Saposhnikoviae extract in water to obtain rehmanniae radix working solution, rhizoma Ligustici Chuanxiong working solution and radix Saposhnikoviae working solution, and mixing the rehmanniae radix working solution, rhizoma Ligustici Chuanxiong working solution and radix Saposhnikoviae working solution to obtain mixed solution I.
4. Mixing
Adding the mixed solution I and the functional auxiliary agent into the residual solvent, mixing to obtain a mixture II, adding the composite gel freeze-dried powder, the human adipose-derived stem cell freeze-dried powder and the human adipose-derived exosomes into the mixture II at 4 ℃, and homogenizing to obtain the stem cell-derived exosome composition for epidermis repair, so that the stem cell-derived exosome composition for epidermis repair finally comprises the following raw materials: 1.0 wt% of composite gel freeze-dried powder, 3.0 wt% of rehmannia root extract, 1.0 wt% of ligusticum wallichii extract, 1.0 wt% of radix sileris extract, 0.6 wt% of exosome derived from human adipose-derived stem cells, the balance of solvent and functional additive, wherein the functional additive is cosolvent and a proper amount of the cosolvent.
Example 5
The present embodiment aims to provide a stem cell-derived exosome composition for epidermal repair, which is mainly prepared by the following steps:
1. preparation of composite gel
Preparing collagen into a 1.2 wt% collagen solution, preparing sodium hyaluronate into a 1.0% sodium hyaluronate solution, mixing the collagen solution and the sodium hyaluronate solution, adding genipin with a final concentration of 0.02mol/L into the mixture, crosslinking the mixture at 45 ℃ for 2.5 hours to obtain a mixture, pre-freezing the mixture, and then freeze-drying the mixture at-80 ℃ for 48 hours to obtain the composite gel freeze-dried powder.
2. Preparation of exosomes
Exosomes were prepared according to the procedure in example 1, with exosomes having a purity of over 96% and a concentration of 2.0g/L being selected.
3. Preparation of plant extract working solution
Respectively dissolving rehmanniae radix extract, rhizoma Ligustici Chuanxiong extract and radix Saposhnikoviae extract in water to obtain rehmanniae radix working solution, rhizoma Ligustici Chuanxiong working solution and radix Saposhnikoviae working solution, and mixing the rehmanniae radix working solution, rhizoma Ligustici Chuanxiong working solution and radix Saposhnikoviae working solution to obtain mixed solution I.
4. Mixing
Adding the mixed solution I and the functional auxiliary agent into the residual solvent, mixing to obtain a mixture II, adding the composite gel freeze-dried powder, the human adipose-derived stem cell freeze-dried powder and the human adipose-derived exosomes into the mixture II at 4 ℃, and homogenizing to obtain the stem cell-derived exosome composition for epidermis repair, so that the stem cell-derived exosome composition for epidermis repair finally comprises the following raw materials: 0.8 wt% of composite gel freeze-dried powder, 2.5 wt% of rehmannia root extract, 1.25 wt% of ligusticum wallichii extract, 1.25 wt% of radix sileris extract and 0.6 wt% of exosome derived from human adipose-derived stem cells, and the balance of solvent and functional aid, wherein the functional aid is cosolvent and a proper amount of the cosolvent.
Example 6
The present embodiment aims to provide a stem cell-derived exosome composition for epidermal repair, which is mainly prepared by the following steps:
1. preparation of composite gel
Preparing collagen into a 1.2 wt% collagen solution, preparing sodium hyaluronate into a 1.0% sodium hyaluronate solution, mixing the collagen solution and the sodium hyaluronate solution, adding genipin with a final concentration of 0.02mol/L into the mixture, crosslinking the mixture at 45 ℃ for 2.5 hours to obtain a mixture, pre-freezing the mixture, and then freeze-drying the mixture at-80 ℃ for 48 hours to obtain the composite gel freeze-dried powder.
2. Preparation of exosomes
Exosomes were prepared according to the procedure in example 1, with exosomes having a purity of over 96% and a concentration of 2.0g/L being selected.
3. Preparation of plant extract working solution
Respectively dissolving rehmanniae radix extract, rhizoma Ligustici Chuanxiong extract and radix Saposhnikoviae extract in water to obtain rehmanniae radix working solution, rhizoma Ligustici Chuanxiong working solution and radix Saposhnikoviae working solution, and mixing the rehmanniae radix working solution, rhizoma Ligustici Chuanxiong working solution and radix Saposhnikoviae working solution to obtain mixed solution I.
4. Mixing
Adding the mixed solution I and the functional auxiliary agent into the residual solvent, mixing to obtain a mixture II, adding the composite gel freeze-dried powder, the human adipose-derived stem cell freeze-dried powder and the human adipose-derived exosomes into the mixture II at 4 ℃, and homogenizing to obtain the stem cell-derived exosome composition for epidermis repair, so that the stem cell-derived exosome composition for epidermis repair finally comprises the following raw materials: 1.0 wt% of composite gel freeze-dried powder, 2.0 wt% of rehmannia root extract, 1.5 wt% of ligusticum wallichii extract, 1.5 wt% of radix sileris extract and 0.6 wt% of exosome derived from human adipose-derived stem cells, and the balance of solvent and functional aid, wherein the functional aid is cosolvent and a proper amount of the cosolvent.
Example 7
The present embodiment aims to provide a stem cell-derived exosome composition for epidermal repair, which is mainly prepared by the following steps:
1. preparation of composite gel
Preparing collagen into a 1.2 wt% collagen solution, preparing sodium hyaluronate into a 1.0% sodium hyaluronate solution, mixing the collagen solution and the sodium hyaluronate solution, adding genipin with a final concentration of 0.02mol/L into the mixture, crosslinking the mixture at 45 ℃ for 2.5 hours to obtain a mixture, pre-freezing the mixture, and then freeze-drying the mixture at-80 ℃ for 48 hours to obtain the composite gel freeze-dried powder.
2. Preparation of exosomes
Exosomes were prepared according to the procedure in example 1, with exosomes having a purity of over 96% and a concentration of 2.0g/L being selected.
3. Preparation of plant extract working solution
Respectively dissolving rehmanniae radix extract, rhizoma Ligustici Chuanxiong extract and radix Saposhnikoviae extract in water to obtain rehmanniae radix working solution, rhizoma Ligustici Chuanxiong working solution and radix Saposhnikoviae working solution, and mixing the rehmanniae radix working solution, rhizoma Ligustici Chuanxiong working solution and radix Saposhnikoviae working solution to obtain mixed solution I.
4. Mixing
Adding the mixed solution I and the functional auxiliary agent into the residual solvent, mixing to obtain a mixture II, adding the composite gel freeze-dried powder, the human adipose-derived stem cell freeze-dried powder and the human adipose-derived exosomes into the mixture II at 4 ℃, and homogenizing to obtain the stem cell-derived exosome composition for epidermis repair, so that the stem cell-derived exosome composition for epidermis repair finally comprises the following raw materials: 1.2 wt% of composite gel freeze-dried powder, 1.5 wt% of rehmannia root extract, 1.75 wt% of ligusticum wallichii extract, 1.75 wt% of radix sileris extract and 0.6 wt% of exosome derived from human adipose-derived stem cells, and the balance of solvent and functional aid, wherein the functional aid is cosolvent and a proper amount of the cosolvent.
Example 8
The present embodiment aims to provide a stem cell-derived exosome composition for epidermal repair, which is mainly prepared by the following steps:
1. preparation of composite gel
Preparing collagen into a 1.2 wt% collagen solution, preparing sodium hyaluronate into a 1.0% sodium hyaluronate solution, mixing the collagen solution and the sodium hyaluronate solution, adding genipin with a final concentration of 0.02mol/L into the mixture, crosslinking the mixture at 45 ℃ for 2.5 hours to obtain a mixture, pre-freezing the mixture, and then freeze-drying the mixture at-80 ℃ for 48 hours to obtain the composite gel freeze-dried powder.
2. Preparation of exosomes
Exosomes were prepared according to the procedure in example 1, with exosomes having a purity of over 96% and a concentration of 2.0g/L being selected.
3. Preparation of plant extract working solution
Respectively dissolving rehmanniae radix extract, rhizoma Ligustici Chuanxiong extract and radix Saposhnikoviae extract in water to obtain rehmanniae radix working solution, rhizoma Ligustici Chuanxiong working solution and radix Saposhnikoviae working solution, and mixing the rehmanniae radix working solution, rhizoma Ligustici Chuanxiong working solution and radix Saposhnikoviae working solution to obtain mixed solution I.
4. Mixing
Adding the mixed solution I and the functional auxiliary agent into the residual solvent, mixing to obtain a mixture II, adding the composite gel freeze-dried powder, the human adipose-derived stem cell freeze-dried powder and the human adipose-derived exosomes into the mixture II at 4 ℃, and homogenizing to obtain the stem cell-derived exosome composition for epidermis repair, so that the stem cell-derived exosome composition for epidermis repair finally comprises the following raw materials: 0.8 wt% of composite gel freeze-dried powder, 1.0 wt% of rehmannia root extract, 2 wt% of ligusticum wallichii extract, 2 wt% of radix sileris extract and 0.5 wt% of exosome derived from human adipose-derived stem cells, and the balance of solvent and functional additive, wherein the functional additive is cosolvent and is in a proper amount.
Comparative example 1
The purpose of this comparative example is to provide a composition without exosomes, containing the following raw materials: 0.8 wt% of composite gel freeze-dried powder, 2.5 wt% of rehmannia root extract, 1.25 wt% of ligusticum wallichii extract, 1.25 wt% of radix sileris extract, and the balance of solvent and functional auxiliary agent, wherein the functional auxiliary agent is cosolvent and is in a proper amount.
Comparative example 2
The purpose of this comparative example is to provide a composition without added plant extracts, containing the following raw materials: 0.8 wt% of composite gel freeze-dried powder, 0.6 wt% of exosome derived from human adipose-derived stem cells, and the balance of solvent and functional additive, wherein the functional additive is cosolvent and is in a proper amount.
Comparative example 3
The purpose of the comparative example is to provide a composition without adding composite gel freeze-dried powder, which comprises the following raw materials: 2.5 wt% of rehmannia root extract, 1.25 wt% of ligusticum wallichii extract, 1.25 wt% of radix sileris extract, 0.8 wt% of human adipose-derived exosome, and the balance of solvent and functional aid, wherein the functional aid is cosolvent and a proper amount is needed.
Comparative example 4
The purpose of this comparative example is to provide a composition containing only solvent and functional adjuvant.
Examples of effects
The purpose of the effect example is to verify the effect of the compositions provided in the foregoing examples and comparative examples in skin repair.
1. Detection of bacteria content
The compositions provided by examples 2-8 and the compositions provided by comparative examples 1-4 are subjected to bacteria content measurement, and the measurement result shows that the colony content of the compositions provided by the examples and the comparative examples meets the industrial regulations.
2. Allergy test
The back of a rabbit for experiment is unhaired, 11 pieces of skin are exposed, each piece is about 1.5cm2, the serial number is 1-11, wherein 1-7 correspond to the composition provided in the embodiment 2-8 in sequence, the serial number is 8-11 correspond to the composition provided in the comparative example 1-4 in sequence, the experiment lasts for two weeks, and whether abnormal skin conditions such as erythema, edema and the like appear at the smearing part is observed every day.
The results show that the compositions provided in both the examples and the comparative examples are non-irritating.
3. Allergy, acne repair and repair of sensitive muscles caused by damaged epidermis
Gathering volunteers with skin in an allergic stage, dividing the volunteers into 11 groups, 10 people in each group, numbering 1-11, wherein 1-7 correspond to the composition provided in examples 2-8 in sequence, and numbering 8-11 correspond to the composition provided in comparative examples 1-4 in sequence, limiting the number of experimenters in the allergic stage, and performing the test by dividing the test into 3 batches, wherein each group in the first two batches is 3 people, and each group in the last batch is 4 people, and testing the repairing effect of the composition on allergy.
The volunteers whose skin is in the acne stage were collected and divided into 11 groups of 10 people each, numbered 1-11, wherein 1-7 correspond to the compositions provided in examples 2-8 in sequence, and 8-11 correspond to the compositions provided in comparative examples 1-4 in sequence, and the repairing effect of the compositions on acne was tested.
Volunteers were recruited to have skin that was susceptible to allergic, redness, dry itching, acne and stinging, 10 individuals per group, numbered 1-11, wherein 1-7 corresponded in sequence to the compositions provided in examples 2-8, and numbers 8-11 corresponded in sequence to the compositions provided in comparative examples 1-4, and the compositions were tested for their effect on healing damaged skin of the substrate.
The test method comprises the following steps: after each group of volunteers used the same procedure for facial cleansing, the composition was applied uniformly to the face 2 times a day, and was continuously applied until the allergy subsided or acne disappeared.
The repairing effects of allergy, acne repair and damaged epidermis on sensitive muscles are shown in table 1:
TABLE 1
As shown by the results in Table 1, the composition provided by the invention not only can effectively resist allergy and epidermis repair after acne occurs, but also can deeply repair skin frequently suffering from allergy, redness, dry itching, acne and stabbing pain after damage, adjust skin barrier and deeply solve the root cause of skin damage.
In summary, the stem cell-derived exosome composition for epidermal repair according to the embodiments of the present invention has the following advantages:
the first effect is that high-purity exosomes derived from human adipose-derived mesenchymal stem cells are collected by adopting a high-end biotechnology and are used as main functional factors to promote the regeneration of collagen of the skin with damaged basement and accelerate the peeling of aged cells of stratum corneum, and meanwhile, the beneficial factors such as EGF contained in the exosomes derived from human adipose cells have better effect on the regeneration of the collagen of the skin compared with the exosomes derived from other sources.
The second effect is that the compound gel freeze-dried powder is added to form gel in a solvent, which is beneficial to moisturizing the epidermis, and is used for cooperating with various functional components in exosomes to act deeper in the skin, regulating the water-oil balance of the skin, dredging hair follicle sebaceous glands and the like, so that the symptoms of skin damage such as allergy, redness, dry itching, acne, stabbing pain and the like are radically solved.
And thirdly, by adding proper amounts of rehmannia root, ligusticum wallichii and divaricate saposhnikovia root, the composition can promote the absorption of beneficial factors in exosomes to act on skin, can contract blood vessels, inhibit the synthesis and secretion of inflammatory factors, and achieve the effects of diminishing inflammation, relieving pain and removing toxicity, so that the overall effect of the composition is better.
The embodiments described above are some, but not all embodiments of the invention. The detailed description of the embodiments of the present invention is not intended to limit the scope of the invention as claimed, but is merely representative of selected embodiments of the invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Claims (10)
1. A stem cell-derived exosome composition for epidermal repair, comprising human adipose-derived exosomes.
2. The stem cell-derived exosome composition for epidermal repair according to claim 1, wherein the exosome is prepared by culturing the collected adipose-derived mesenchymal stem cells normally until the cell fusion degree reaches 70-80%, replacing the culture medium, continuously culturing for 10-14 h, collecting the supernatant, and extracting by using an exosome extraction kit to obtain the human adipose-derived exosome.
3. The stem cell-derived exosome composition for epidermal repair according to claim 2, further comprising a composite gel lyophilized powder, wherein the composite gel is prepared by cross-linking and lyophilizing sodium hyaluronate and collagen.
4. The stem cell-derived exosome composition for epidermal repair according to claim 3, wherein the composite gel lyophilized powder is prepared by preparing collagen into a collagen solution, preparing sodium hyaluronate into a sodium hyaluronate solution, mixing the collagen solution and the sodium hyaluronate solution, adding genipin into the mixture, crosslinking for 2.5 hours at 45 ℃ to obtain a mixture, pre-freezing the mixture, and then freeze-drying at-80 ℃ to obtain the composite gel lyophilized powder.
5. The stem cell-derived exosome composition for epidermal repair according to claim 3, wherein the concentration of the collagen solution is 1.2 wt%, the concentration of the sodium hyaluronate solution is 1.0 wt%, the volume ratio of the collagen solution to the sodium hyaluronate solution is 1:2, and the final concentration of genipin in the mixture is 0.02 mol/L.
6. The stem cell-derived exosome composition for epidermal repair according to claim 4, wherein the time of freeze-drying is 48 h.
7. The stem cell-derived exosome composition for epidermal repair according to claim 5, further comprising a rehmannia glutinosa extract, a ligusticum chuanxiong hort extract and a saposhnikovia divaricata root extract.
8. The stem cell-derived exosome composition for epidermis repair according to claim 7, wherein the raw materials comprise 0.8-1.2 wt% of composite gel lyophilized powder, 1.0-4.0 wt% of rehmannia root extract, 0.5-2.0 wt% of ligusticum wallichii extract, 0.5-2.0 wt% of radix sileris extract, 0.5-0.8 wt% of human adipose stem cell-derived exosome, and the balance of solvent and functional assistant.
9. A method for preparing a stem cell-derived exosome composition for epidermal repair according to claim 8, comprising the steps of: respectively dissolving the rehmannia root extract, the ligusticum wallichii extract and the radix sileris extract in partial solvents, mixing to obtain a mixed solution I, adding the mixed solution I and the functional auxiliary agent into the residual solvent, mixing to obtain a mixture II, adding the composite gel freeze-dried powder, the human adipose mesenchymal stem cell freeze-dried powder and the human adipose stem cell-derived exosome at the temperature of 4 ℃, and homogenizing to obtain the stem cell-derived exosome composition for epidermis repair.
10. Use of the stem cell-derived exosome composition for epidermal repair according to any one of claims 1 to 7 in the preparation of a skin care and/or cosmetic product.
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Application publication date: 20210910 |