A skin repair composition capable of increasing cell migration rate and a preparation method thereof

Abstract

The present invention belongs to the field of A61K cosmetics technology, and more specifically relates to a skin repair composition capable of improving cell migration and its preparation method. The composition comprises, by weight, 0.01-1% collagen, 0.004-0.5% fibronectin, 1-5% plant extract, and a solvent to make up the remainder. The composition provided by the present invention is safe for the human body, has no side effects, contains few ingredients, and is suitable for use on all skin types. It fundamentally addresses skin repair and can be promoted and used in skin care products such as moisturizers, cleansers, eye creams, sunscreens, and facial masks.

Description

Technical Field

[0001] The present invention belongs to the technical field of A61K cosmetics, and more specifically relates to a skin repair composition capable of improving cell migration rate and a preparation method thereof. Background Art

[0002] Cell migration is the movement of cells after they sense certain substances on their surface. On the skin surface, this can manifest as skin repair and wound healing. In recent years, with the rapid development of the cosmetics industry, skin care products related to skin repair have become increasingly diverse and complex.

[0003] The Chinese invention patent with patent application number 202010455462.8 discloses a repair composition, a repair mask liquid and a repair mask. In the disclosed patent, a repair composition is prepared by using the synergistic effect of nexin, creatine, 1-methylhydantoin-2-imide, and oligopeptide-1. In addition, a mask containing this composition is also studied in the disclosed patent. However, this composition has relatively more ingredients, which will not only greatly increase the cost, but also, because of the presence of creatine, if it cannot be quickly utilized in the skin, it may cause certain damage to the skin over time.

[0004] At present, most skin repair products are limited to achieving skin repair effects through better absorption on the skin surface. However, there are relatively few studies on improving cell migration rate. Improving skin repair by improving cell migration rate is the key to fundamentally solving skin problems. Therefore, the present invention will focus on exploring how to improve cell migration rate and promote skin repair. Summary of the Invention

[0005] In order to solve the above technical problems, the first aspect of the present invention provides a skin repair composition capable of improving cell migration rate, wherein the raw materials for preparation include, by weight percentage:

[0006] Collagen 0.01-10%;

[0007] Fibronectin 0.004-5%;

[0008] Plant extract 1-5%;

[0009] The solvent was added to make up the balance to 100%.

[0010] In some preferred embodiments, the plant extract is selected from Cryptomeria japonica bud extract and / or Cryptomeria japonica leaf plant extract.

[0011] Further preferably, the plant extract includes Cryptomeria japonica bud extract.

[0012] Cryptomeria japonica bud extract: INCI: CRYPTOMERIA JAPONICA BUD EXTRACT.

[0013] In some preferred embodiments, the collagen is selected from at least one of recombinant human collagen and traditional collagen.

[0014] More preferably, the collagen is recombinant human collagen.

[0015] Collagen is an essential component of the skin, maintaining its elasticity and youthfulness. When collagen metabolism outpaces its synthesis, it can cause skin aging. To further maintain the skin's youthful appearance and enhance its ability to repair external pollution and internal damage, supplementing the skin with the appropriate amount of collagen is a crucial and necessary task.

[0016] However, traditional practices mostly use traditional collagen, which is mainly purified from animal tissues. Although it has higher active recognition of basic tripeptides (GER, GEK), because it comes from animal tissues, it may face the risk of infection from some common animal diseases. Not only will it fail to play its original role in repairing and protecting the skin, it will also cause additional damage to the skin.

[0017] Therefore, during a large amount of research, the applicant found that the use of recombinant human collagen in the preparation of the present system composition ensures better cell migration effect and enhances the repair effect on the skin. The applicant speculates that the possible reason is that: recombinant human collagen can be significantly expressed in LOR and TGM1 skin barrier-related genes, promoting the repair of keratin and ensuring the integrity of keratin, which can further provide barrier protection for the skin. In addition, it also shows high expression in the cytoskeletal protein gene K16, which promotes the migration of skin surface cells and ensures the rapid repair of damaged skin.

[0018] In some preferred embodiments, the weight ratio of the collagen to the plant extract is (0.01-10): (1-5).

[0019] In some preferred embodiments, the weight ratio of the collagen to the plant extract is 1:20.

[0020] During the experiment, the applicant discovered through a large number of creative experimental explorations that in this system, through the interaction between collagen and plant extracts, the cell migration rate during the use of the composition can be further improved, especially when the weight ratio of collagen and plant extract is (0.01-10): (1-5), better results are shown. The applicant speculates that the reason for this phenomenon may be because: the collagen added to this system has a triple helical structure, which can provide an attachment site for the cedar bud extract, ensuring better fusion between the plant extract and the collagen, and thus ensuring that it can form a stable release body between the raw materials for preparing skin care products during subsequent use, ensuring that the active ingredients can exert their effects.

[0021] In addition, the applicant has found that if the content of plant extract added to the system is too high, it may cause it to become oversaturated, thereby causing the loss of effective ingredients, which not only increases costs but also causes the cell migration rate to decrease instead of increase.

[0022] In some preferred embodiments, the fibronectin is at least one selected from human fibronectin, bovine fibronectin, monkey fibronectin, rabbit fibronectin, mouse fibronectin, and salmon fibronectin.

[0023] More preferably, the fibronectin is human fibronectin.

[0024] More preferably, the average molecular weight of the human fibronectin is 10,000-50,000.

[0025] More preferably, the average molecular weight of the human fibronectin is 38,000.

[0026] Human fibronectin, product number MELLPRO 500G, was purchased from Meierjian (Shenzhen) Biotechnology Co., Ltd.

[0027] In this system, in the presence of collagen and plant extracts, the addition of human fibronectin with an average molecular weight of 38,000 can form a uniform and stable mixed state. Due to the presence of a certain molecular weight of fibronectin, a certain fusion state can be provided for this system. When used on human skin, it can form a stable attachment state on the skin surface, facilitating the skin's absorption of the active ingredients in the composition. It can also enable cells to sense the active substances and initiate movement, thereby promoting the improvement of cell migration rate and promoting skin repair.

[0028] In addition, if the average molecular weight of fibronectin is too small during use, it may lead to a decrease in its adhesion to the skin surface during use, thereby causing a decrease in the utilization rate of active ingredients, affecting its cell migration rate, and thus affecting its application in the field of skin care products.

[0029] In some preferred embodiments, the solvent includes but is not limited to 1,4-butanediol, 1,3-propylene glycol, and water.

[0030] More preferably, the solvent includes 1,4-butanediol and water.

[0031] More preferably, the raw materials for preparing the composition include, by weight percentage:

[0032] Collagen 0.01-10%;

[0033] Fibronectin 0.004-5%;

[0034] Plant extract 1-5%;

[0035] 1,4-Butanediol 25-70%;

[0036] Add water to make up to 100%.

[0037] In some preferred embodiments, the present invention also provides an application of a skin repair composition capable of improving cell migration rate, which is used in moisturizing creams, cleansing milks, eye creams, sunscreens, and facial masks.

[0038] In some preferred embodiments, the present invention further provides a skin care product comprising the skin repair composition of the present invention.

[0039] In some preferred embodiments, 0.1-5 parts of the skin repair composition are contained in every 100 parts by weight of the skin care product.

[0040] Another aspect of the present invention provides a method for preparing a skin repair composition capable of improving cell migration rate, comprising the following steps:

[0041] The prepared raw materials are mixed, stirred and homogenized to obtain the product.

[0042] Beneficial effects: The skin repair composition capable of improving cell migration provided by the present invention has the following advantages over the prior art:

[0043] 1. The composition prepared by the present invention has a protective effect on the skin. During use, it can form a barrier on the surface layer of the skin to prevent the invasion of harmful substances;

[0044] 2. The composition prepared by the present invention has the advantages of low dosage and good effect. During use, it can have good adhesion on the skin surface, ensuring that the active substance is absorbed by the skin surface cells and senses the movement signal, thereby promoting skin repair and wound healing;

[0045] 3. The composition provided by the present invention is safe to the human body without any side effects, has few ingredients, is suitable for all skin types, and fundamentally solves the problem of skin repair. It can be promoted and used in skin care products such as moisturizing creams, facial cleansers, eye creams, sunscreens, and facial masks. BRIEF DESCRIPTION OF THE DRAWINGS

[0046] Figure 1 This is the cell migration diagram of the blank control group at 0h and 24h in Example 1 of this application;

[0047] Figure 2 This is the cell migration diagram of the positive control group at 0h and 24h in Example 1 of the present application;

[0048] Figure 3 This is the cell migration diagram of fibronectin at 0h and 24h in the composition of Example 1 of the present application;

[0049] Figure 4 This is the cell migration diagram of collagen at 0h and 24h in the composition of Example 1 of the present application;

[0050] Figure 5 This is a cell migration diagram of the plant extract in the composition of Example 1 of the present application at 0h and 24h. DETAILED DESCRIPTION

[0051] Example

[0052] Example 1

[0053] A skin repair composition capable of improving cell migration rate, the raw materials for preparation comprising, by weight:

[0054] Collagen 0.1%;

[0055] Fibronectin 0.01%;

[0056] Plant extract 2%;

[0057] 1,4-Butanediol 65%;

[0058] Add water to make up to 100%.

[0059] The collagen is recombinant human collagen, purchased from Jiangsu Chuangjian Medical Technology Co., Ltd.

[0060] Human fibronectin, with an average molecular weight of 38,000 and product number MELLPRO 500G, was purchased from Meierjian (Shenzhen) Biotechnology Co., Ltd.

[0061] The plant extract is Cryptomeria japonica bud extract, INCI: CRYPTOMERIA JAPONICA BUD EXTRACT, purchased from Gattefossé (Shanghai) Trading Co., Ltd.

[0062] A method for preparing a skin repair composition capable of improving cell migration rate comprises the following steps:

[0063] The prepared raw materials are mixed, stirred and homogenized to obtain the product.

[0064] Example 2

[0065] A skin repair composition capable of improving cell migration rate, the raw materials for preparation comprising, by weight:

[0066] Collagen 0.01%;

[0067] Fibronectin 0.004%;

[0068] Plant extract 1%;

[0069] 1,4-Butanediol 70%;

[0070] Add water to make up to 100%.

[0071] The collagen is recombinant human collagen, purchased from Jiangsu Chuangjian Medical Technology Co., Ltd.

[0072] Human fibronectin, with an average molecular weight of 38,000 and product number MELLPRO 500G, was purchased from Meierjian (Shenzhen) Biotechnology Co., Ltd.

[0073] The plant extract is Cryptomeria japonica bud extract, INCI: CRYPTOMERIA JAPONICA BUD EXTRACT, purchased from Gattefossé (Shanghai) Trading Co., Ltd.

[0074] A method for preparing a skin repair composition capable of improving cell migration rate comprises the following steps:

[0075] The prepared raw materials are mixed, stirred and homogenized to obtain the product.

[0076] Example 3

[0077] A skin repair composition capable of improving cell migration rate, the raw materials for preparation comprising, by weight:

[0078] Collagen 10%;

[0079] Fibronectin 5%;

[0080] Plant extract 5%;

[0081] 1,4-Butanediol 60%;

[0082] Add water to make up to 100%.

[0083] The collagen is recombinant human collagen, purchased from Jiangsu Chuangjian Medical Technology Co., Ltd.

[0084] Human fibronectin, with an average molecular weight of 38,000 and product number MELLPRO 500G, was purchased from Meierjian (Shenzhen) Biotechnology Co., Ltd.

[0085] The plant extract is Cryptomeria japonica bud extract, INCI: CRYPTOMERIA JAPONICA BUD EXTRACT, purchased from.

[0086] A method for preparing a skin repair composition capable of improving cell migration rate comprises the following steps:

[0087] The prepared raw materials are mixed, stirred and homogenized to obtain the product.

[0088] Example 4

[0089] A skin repair composition capable of improving cell migration rate, wherein the specific implementation manner is the same as that of Example 1, but the difference from Example 1 is that the composition does not contain fibronectin.

[0090] The collagen is recombinant human collagen, purchased from Jiangsu Chuangjian Medical Technology Co., Ltd.

[0091] The plant extract is Cryptomeria japonica bud extract, INCI: CRYPTOMERIA JAPONICA BUD EXTRACT, purchased from Gattefossé (Shanghai) Trading Co., Ltd.

[0092] A method for preparing a skin repair composition capable of improving cell migration rate comprises the following steps:

[0093] The prepared raw materials are mixed, stirred and homogenized to obtain the product.

[0094] Comparative Example 1

[0095] A skin repair composition capable of improving cell migration rate, the specific implementation of which is the same as that of Example 1, except that the fibronectin is human fibronectin with an average molecular weight of 220,000, purchased from Shanghai Qianchen Biotechnology Co., Ltd.

[0096] A method for preparing a skin repair composition capable of improving cell migration rate comprises the following steps:

[0097] The prepared raw materials are mixed, stirred and homogenized to obtain the product.

[0098] Comparative Example 2

[0099] A skin repair composition capable of improving cell migration rate, wherein the specific implementation manner is the same as that of Example 1, but the difference from Example 1 is that the composition does not contain plant extracts.

[0100] A method for preparing a skin repair composition capable of improving cell migration rate comprises the following steps:

[0101] The prepared raw materials are mixed, stirred and homogenized to obtain the product.

[0102] Performance testing:

[0103] 1. Cell migration rate test:

[0104] Test system: Keratinocytes: Ep200708, P7; Fibroblasts: FB190520; 3D epidermal skin model ES210703 was provided by Guangdong Boxi Biotechnology Co., Ltd.

[0105] 2. Test reagents:

[0106] MTT (Sigma), KcGrowth (Boxi, Guangdong), EpiGrowth culture medium (Boxi, Guangdong), PBS (Solaibao), SLS (Sigma), 4% paraformaldehyde (BioSharp), antifade agent (Biyuntian), Hochest33342 (Biyuntian), ready-to-use goat serum (Zhongshan Jinqiao).

[0107] 3. Test equipment:

[0108] CO2 incubator (Thermo, 150i), clean bench (Suzhou Antai, SW-CJ-1F), microplate reader (BioTek, Epoch), fluorescence microscope (Leica; DM2500), upright microscope (Olympus, BX53).

[0109] 4. Cell scratch test:

[0110] 3.1 Cell scratch assay

[0111] The sample's promotion of cell migration illustrates the sample's repair effect based on keratinocytes.

[0112] 1) Cell seeding: Keratinocytes were seeded into 6-well plates at a seeding density of 4.5E5 cells / well and incubated overnight in an incubator (37° C., 5% CO 2 ).

[0113] 2) Prepare the sample working solution according to the experimental design (Table 1).

[0114] Table 1 Experimental design

[0115]

[0116]

[0117] 3) Dosing: When the cell plating rate in the 6-well plate reaches approximately 80%, group dosing is performed according to the experimental design (Table 1). The dosage per well is 2 mL, and three replicate wells are set up for each group. Culture is continued in an incubator (37°C, 5% CO2) for 24 hours.

[0118] 4) Scratching: After the sample is incubated, scratch it.

[0119] 5) Washing: After the scratching, the cells were washed three times with PBS, added with cell culture medium, and cultured in an incubator (37° C., 5% CO 2 ) for 24 h.

[0120] 6) Photography: Take photos under a 4x microscope 0h and 24h after scratching.

[0121] 3.2 Scratch test results

[0122] Based on the experimental method, the migration rate of the solvent control (SC) group was normalized and the relative migration rate of each group was calculated. The cell migration was shown below. Figure 1-Figure 5 , the data analysis results are shown in Table 2.

[0123] Table 2

[0124]

[0125]

[0126] 2. Barrier index test:

[0127] Barrier Index ET 50 , meaning Triton X-100 is moderately irritating to the skin and can cause barrier damage upon skin contact. When Triton X-100 is applied to the model, cell viability will gradually decrease over time. When the model, after exposure to the sample, is stimulated with Triton X-100, the differences in barrier efficacy between the samples will gradually increase over time. By comparing the differences in viability between the sample and control groups, the differences in barrier efficacy caused by the samples can be assessed and indirectly predicted.

[0128] 1) Preparation of working fluid

[0129] a. Preparation of 0.2% SLS working solution: Pipette 1 mL of 0.4% SLS solution and add 1 mL of PBS to prepare 0.2% SLS working solution.

[0130] Barrier index ET50 (detection model: EpiKutis)

[0131] Detection method: Use SLS to stimulate the epidermal skin model EpiKutis to construct an in vitro skin damage model. By detecting the tissue morphology of the skin model after the sample is exposed to the sample, the repair efficacy of the sample to be tested is evaluated.

[0132] 4.4 Barrier Index ET50 Test Results

[0133] According to the specific test method, the composition prepared in Example 1 was subjected to tissue viability testing, and the results were as follows:

[0134] experiment Time point setting (h) <![CDATA[ET 50 (h)]]> Example 1 0，1，3 2.76

[0135] 3. The compositions prepared in Examples 1-3 and Comparative Examples 1-2 were subjected to stability testing. They were placed at 54° C. for one week, and the changes in the surface were observed. The results are recorded in the table below.

[0136]

[0137]

Claims

1. A skin repair composition capable of improving cell migration rate, characterized in that: The raw materials for preparation include, by weight: Collagen 0.1% Fibronectin 0.01% Plant extract 2% 1,4-Butanediol 65% Add water to 100% of the balance; The collagen is recombinant human collagen; Human fibronectin, average molecular weight 38,000, product number MELLPRO 500G; The plant extract is a cedar bud extract.

2. A skin care product, characterized in that: The invention comprises the skin repairing composition according to claim 1.

3. The skin care product according to claim 2, characterized in that The skin care product contains 0.1-5 parts of the skin repair composition according to claim 1 per 100 parts by weight.

4. A method for preparing the skin repair composition capable of improving cell migration rate according to claim 1, characterized in that: The following steps are involved: The prepared raw materials are mixed, stirred and homogenized to obtain the product.

Images