Facial mask containing stem cell secretion active peptide and preparation process thereof

Abstract

The invention provides a facial mask containing stem cell secreted active peptides and a preparation process of the facial mask. According to the invention, bone marrow mesenchymal stem cells are used as starting cells, main secretory active peptides are prepared by means of ultrafiltration, enzymolysis and the like, and the mask disclosed by the invention is constructed by using the secretory active peptides. Results show that the mask added with the specific stem cell secreted active peptide not only improves the problem of dry skin, but also can effectively recover skin elasticity and reduce wrinkles, has a good antibacterial effect, can effectively inhibit propionibacterium sores and malassezia, and has a good acne treatment effect.

Description

Technical Field

[0001] The present invention relates to the technical field of facial masks, and particularly to a facial mask containing stem cell-secreted active peptides and a preparation process thereof. Background Art

[0002] Facial masks are a type of cosmetic that has been used for a long time. As early as the era of the ancient Egyptian pyramids, it was known to use some natural raw materials, such as soil, volcanic ash, sea mud, etc., to apply on the face or body to treat some skin diseases. Later, it developed to mix lanolin with various substances such as honey, flower classes of plants, eggs, coarse flour, coarse beans, etc., and make it into a paste, which is applied on the face for beauty or treatment of some skin diseases.

[0003] However, the quality of existing facial mask products varies greatly, and the claimed effects are often false propaganda, resulting in certain selection obstacles for users in purchasing facial masks. Moreover, the functions of existing facial masks are too single, and no facial mask that combines multiple functions has emerged. In view of the current situation of facial masks, the present invention creatively proposes the following technical solutions. Summary of the Invention

[0004] Aiming at the technical problems existing in the prior art, the present invention provides a facial mask containing stem cell-secreted active peptides and a preparation process thereof. The present invention uses bone marrow mesenchymal stem cells as starting cells, and prepares the main secreted active peptides through means such as ultrafiltration and enzymatic hydrolysis, and constructs the facial mask of the present invention with these.

[0005] Specifically, the present invention first provides a facial mask containing stem cell-secreted active peptides, which is characterized in that the facial mask is composed of 10-20 parts of bone marrow mesenchymal stem cell-secreted active peptides prepared by the above method, 50-80 parts of cucumber juice, 5-10 parts of glycerol, 0.1-0.5 part of vitamin C, 5-10 parts of hyaluronic acid, and 50-100 parts of ionized water.

[0006] Preferably, the bone marrow mesenchymal stem cell-secreted active peptides are selected from any one or a combination of several of SEQ ID NO.1-3.

[0007] Preferably, the facial mask containing stem cell-secreted active peptides is characterized in that the facial mask is composed of 10 parts of bone marrow mesenchymal stem cell-secreted active peptides prepared by the above method, 50 parts of cucumber juice, 5 parts of glycerol, 0.1 part of vitamin C, 5 parts of hyaluronic acid, and 100 parts of ionized water. Among them, the sequence of the bone marrow mesenchymal stem cell-secreted active peptides is as shown in SEQ ID NO.1.

[0008] Preferably, the mask containing the stem cell-secreted active peptide is characterized in that the mask is composed of 10 parts of the active peptide secreted by bone marrow mesenchymal stem cells, 50 parts of cucumber juice, 5 parts of glycerol, 0.1 part of vitamin C, 5 parts of hyaluronic acid, and 100 parts of ionized water prepared by the above method. Among them, the sequence of the active peptide secreted by bone marrow mesenchymal stem cells is shown in SEQ ID NO.2.

[0009] Preferably, the mask containing the stem cell-secreted active peptide is characterized in that the mask is composed of 10 parts of the active peptide secreted by bone marrow mesenchymal stem cells, 50 parts of cucumber juice, 5 parts of glycerol, 0.1 part of vitamin C, 5 parts of hyaluronic acid, and 100 parts of ionized water prepared by the above method. Among them, the sequence of the active peptide secreted by bone marrow mesenchymal stem cells is shown in SEQ ID NO.3.

[0010] On the other hand, the present invention also provides a method for preparing the mask, which includes the following steps:

[0011] 1) Take 50 - 100 parts of deionized water, and sequentially add 5 - 10 parts of glycerol and 5 - 10 parts of hyaluronic acid, and dissolve them fully in a water bath environment at 25 - 37 °C;

[0012] 2) Sequentially add 10 - 20 parts of the active peptide secreted by bone marrow mesenchymal stem cells and 0.1 - 0.5 part of vitamin C to step 1), and dissolve them fully in a water bath environment at 20 - 30 °C;

[0013] 3) Add 50 - 80 parts of cucumber juice to the solution in step 2), and slowly stir until it is fully dissolved to obtain the mask.

[0014] Preferably, the method for preparing the mask includes the following steps:

[0015] 1) Take 100 parts of deionized water, and sequentially add 5 parts of glycerol and 5 parts of hyaluronic acid, and dissolve them fully in a water bath environment at 25 °C;

[0016] 2) Sequentially add 10 parts of the active peptide secreted by bone marrow mesenchymal stem cells and 0.1 part of vitamin C to step 1), and dissolve them fully in a water bath environment at 25 °C;

[0017] 3) Add 50 parts of cucumber juice to the solution in step 2), and slowly stir until it is fully dissolved to obtain the mask.

[0018] Further preferably, the use of the active peptide secreted by bone marrow mesenchymal stem cells in the preparation of products for resisting skin aging, wrinkle removal and firming, or treating skin acne.

[0019] Further preferably, the product further includes common excipients in cosmetics.

[0020] The advantages of the present invention are as follows: The present invention uses bone marrow mesenchymal stem cells as starting cells, and prepares the main secreted active peptides through means such as ultrafiltration and enzymatic hydrolysis, and constructs the facial mask of the present invention with these. The results show that the facial mask of the present invention adding specific stem cell-secreted active peptides not only improves the problem of dry skin, but also can effectively restore skin elasticity, reduce the generation of wrinkles. In addition, it has a good antibacterial effect, can effectively inhibit Propionibacterium acnes and Malassezia, and has a good acne treatment effect. Detailed implementation manners

[0021] The following combines specific embodiments to further elaborate on the present invention in detail, so that those skilled in the art can understand the present invention more clearly.

[0022] The following embodiments are only used to illustrate the present invention and are not used to limit the scope of the present invention. Based on the specific embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative work belong to the protection scope of the present invention.

[0023] In the embodiments of the present invention, unless otherwise specified, all raw material components are commercially available products well-known to those skilled in the art; in the embodiments of the present invention, if not specifically specified, the technical means used are all conventional means well-known to those skilled in the art.

[0024] Example 1

[0025] Extraction method of bone marrow mesenchymal stem cell-secreted active peptides, the method comprising the following steps:

[0026] 1) Take bone marrow tissue for trypsin digestion to prepare a single cell suspension, and use a flow cytometer to sort bone marrow mesenchymal stem cells of + CD29 + CD44 + CD73 + CD166 - CD14 - CD34 - CD45;

[0027] 2) Place the bone marrow mesenchymal stem cells obtained in step 1) in DMEM / F12 medium for continuous passage culture;

[0028] 3) After it is stable after three passages, collect the supernatant of the fourth passage of bone marrow mesenchymal stem cells, filter and sterilize it with a 0.22 μm filter membrane, and perform secondary filtration with an ultrafiltration membrane with a molecular weight cut-off of 20 KD;

[0029] 4) Add 150 U / L collagenase to the secondary filtrate in step 3), enzymatically hydrolyze at 37 °C for 2 h; after enzymatic hydrolysis, inactivate the enzyme, and centrifuge at 5000 g / min to separate the supernatant for standby;

[0030] 5) Filtration and concentration: The supernatant from step 4) is filtered using an ultrafiltration membrane, and the polypeptide molecules with a molecular weight cut-off of 1000 - 3500 Da are retained for further use.

[0031] 6) Identification of bioactive peptides: The concentrated solution from step 5) is subjected to HPLC and sequencing using an Obitrap Fusion Lumos mass spectrometer to identify the bioactive peptides, and the bioactive peptide samples are collected for further use.

[0032] 7) Vacuum drying: The bioactive peptides identified in step 6) are vacuum concentrated to a concentration of 18° Bé and spray-dried into fine powder for further use.

[0033] Through HPLC and mass spectrometer identification and analysis, a total of three main secreted bioactive peptides are obtained, and their sequences are shown as SEQ ID NO.1 - 3 in sequence.

[0034] Example 2

[0035] A facial mask containing stem cell-secreted bioactive peptides, characterized in that the facial mask is composed of 10 parts of bone marrow mesenchymal stem cell-secreted bioactive peptides, 50 parts of cucumber juice, 5 parts of glycerol, 0.1 part of vitamin C, 5 parts of hyaluronic acid, and 100 parts of ionized water prepared by the above method.

[0036] Among them, the sequence of the bone marrow mesenchymal stem cell-secreted bioactive peptide is as shown in SEQ ID NO.1.

[0037] The preparation method of the facial mask includes the following steps:

[0038] 1) In 100 parts of deionized water, 5 parts of glycerol and 5 parts of hyaluronic acid are added in sequence, and they are fully dissolved in a water bath environment at 25°C.

[0039] 2) 10 parts of bone marrow mesenchymal stem cell-secreted bioactive peptides and 0.1 part of vitamin C are added to step 1) in sequence, and they are fully dissolved in a water bath environment at 25°C.

[0040] 3) 50 parts of cucumber juice are added to the solution in step 2), and it is slowly stirred until fully dissolved to obtain the facial mask.

[0041] Example 3

[0042] A facial mask containing stem cell-secreted bioactive peptides, characterized in that the facial mask is composed of 10 parts of bone marrow mesenchymal stem cell-secreted bioactive peptides, 50 parts of cucumber juice, 5 parts of glycerol, 0.1 part of vitamin C, 5 parts of hyaluronic acid, and 100 parts of ionized water.

[0043] Among them, the sequence of the bone marrow mesenchymal stem cell-secreted bioactive peptide is as shown in SEQ ID NO.2.

[0044] The preparation method is the same as that of Example 2.

[0045] Example 4

[0046] The facial mask containing the stem cell-secreted bioactive peptide is characterized in that the facial mask is composed of 10 parts of the bioactive peptide secreted by bone marrow mesenchymal stem cells, 50 parts of cucumber juice, 5 parts of glycerol, 0.1 part of vitamin C, 5 parts of hyaluronic acid and 100 parts of ionized water.

[0047] Among them, the sequence of the bioactive peptide secreted by the bone marrow mesenchymal stem cells is as shown in SEQ ID NO.3.

[0048] The preparation method is the same as that of Example 2.

[0049] Control Example

[0050] The preparation method steps of Control Example 1 are the same as those of Example 2, except that the active ingredient of the facial mask does not contain the bioactive peptide secreted by bone marrow mesenchymal stem cells, but uses an equal amount of deionized water as a supplement. The preparation method is the same as that of Example 2.

[0051] Test Example 1

[0052] Test on the improvement of facial skin condition: 100 testers with dry and water-deficient skin, with an average age of 35 years old, were selected. The testers were successively divided into a blank group, Experimental Groups 1-3 and Control Group 1. Among them, in the blank group, no facial mask was applied in the morning, noon and evening, and only regular facial cleansing was done. The testers in the experimental groups applied the corresponding facial masks every night. After continuous application for 7 days, the skin conditions of the testers before and after the test were tested with a facial image analyzer VISIA-CR and a multi-functional skin tester CK-MPA10. The results are shown in Table 1.

[0053] Table 1 Changes in various facial skin indexes before and after using the facial mask for 7 days

[0054] Group Facial oil rate / % Facial moisture / % Facial elasticity / % Facial wrinkles / % Blank group 50 5 10 25 Experimental group 1 5 50 35 10 Experimental group 2 8 45 30 15 Experimental group 3 10 47 32 20 Control group 1 55 4 8 3

[0055] The detection results are as shown in Table 1 above: After continuous monitoring of the testers in each group for 7 days, the skin oil content of the testers in the experimental groups decreased, and the water content increased significantly; and the skin elasticity was effectively restored, and the number of facial wrinkles also decreased significantly, fully indicating that the facial mask of the present invention has good anti-aging, water retention and firming deposition effects.

[0056] Test Example 2

[0057] Analysis of the antibacterial effect of the facial mask: Propionibacterium acnes and Malassezia are the main pathogenic bacteria causing skin acne. To further verify the antibacterial ability of the facial mask of the present invention, referring to the antibacterial performance test method for dissolving antibacterial (or inhibiting) products in Appendix C4 of "GB15979-2002 Hygiene Standards for Disposable Sanitary Products", the inhibition rates of the facial mask liquids of Examples 2-4 and the comparative examples against Propionibacterium acnes (ATCC11827) and Malassezia (ATCC 44344) were tested. The results are shown in Table 2 below.

[0058] Table 2. Inhibition rates of facial mask liquid against Propionibacterium acnes and Malassezia

[0059] Group Propionibacterium acnes / % Malassezia / % Blank group 100 100 Experimental group 1 25 15 Experimental group 2 36 29 Experimental group 3 40 32 Control group 1 96 95

[0060] The above data confirm that: the facial mask of the present invention has a good inhibitory effect on Propionibacterium acnes and Malassezia, further confirming that the facial mask of the present invention has a good acne treatment effect.

[0061] It is necessary to point out here that the above embodiments are only for further elaboration and explanation of the technical solution of the present invention, and are not a further limitation on the technical solution of the present invention. The method of the present invention is only a preferred implementation scheme and is not used to limit the protection scope of the present invention. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included within the protection scope of the present invention.

Claims

1. Bone marrow mesenchymal stem cells secrete active peptides, characterized in that, The amino acid sequence is any one or a combination of several of those shown in SEQ ID NO.1-3.

2. A facial mask containing stem cell-secreted active peptides, characterized in that, The facial mask comprises 10-20 parts of active peptides secreted by bone marrow mesenchymal stem cells, 50-80 parts of cucumber juice, 5-10 parts of glycerol, 0.1-0.5 part of vitamin C, 5-10 parts of hyaluronic acid and 50-100 parts of ionized water. The active peptides secreted by bone marrow mesenchymal stem cells are selected from any one or a combination of several of those shown in SEQ ID NO.1-3.

3. The facial mask according to claim 2, wherein, The facial mask comprises 10 parts of active peptides secreted by bone marrow mesenchymal stem cells, 50 parts of cucumber juice, 5 parts of glycerol, 0.1 part of vitamin C, 5 parts of hyaluronic acid and 100 parts of ionized water. Among them, the sequence of the active peptides secreted by bone marrow mesenchymal stem cells is as shown in SEQ ID NO.

1.

4. The facial mask according to claim 2, wherein, The facial mask comprises 10 parts of active peptides secreted by bone marrow mesenchymal stem cells, 50 parts of cucumber juice, 5 parts of glycerol, 0.1 part of vitamin C, 5 parts of hyaluronic acid and 100 parts of ionized water. Among them, the sequence of the active peptides secreted by bone marrow mesenchymal stem cells is as shown in SEQ ID NO.

2.

5. The facial mask according to claim 2, wherein The facial mask comprises 10 parts of active peptides secreted by bone marrow mesenchymal stem cells, 50 parts of cucumber juice, 5 parts of glycerol, 0.1 part of vitamin C, 5 parts of hyaluronic acid and 100 parts of ionized water. Among them, the sequence of the active peptides secreted by bone marrow mesenchymal stem cells is as shown in SEQ ID NO.

3.

6. The preparation method of the facial mask according to any one of claims 2-5, characterized in that Comprising the following steps: 1) Take 50-100 parts of deionized water, and successively add 5-10 parts of glycerol and 5-10 parts of hyaluronic acid, and fully dissolve them in a water bath environment at 25-37°C. 2) Successively add 10-20 parts of active peptides secreted by bone marrow mesenchymal stem cells and 0.1-0.5 part of vitamin C to step 1), and fully dissolve them in a water bath environment at 20-30°C. 3) Add 50-80 parts of cucumber juice to the solution in step 2), and slowly stir until it is fully dissolved to obtain the product.

7. The preparation method of the facial mask according to claim 6, characterized in that, Comprising the following steps: 1) Take 100 parts of deionized water, and successively add 5 parts of glycerol and 5 parts of hyaluronic acid, and fully dissolve them in a water bath environment at 25°C. 2) Successively add 10 parts of active peptides secreted by bone marrow mesenchymal stem cells and 0.1 part of vitamin C to step 1), and fully dissolve them in a water bath environment at 25°C. 3) Add 50 parts of cucumber juice to the solution in step 2), and slowly stir until it is fully dissolved to obtain the product.

8. Use of the active peptides secreted by the bone marrow mesenchymal stem cells as claimed in claim 1 in the preparation of a product for resisting skin aging or a product for skin wrinkle removal and firming or a product for treating skin acne.