A basement membrane repair composition and its preparation method and application

Abstract

The present invention discloses a basement membrane repair composition and its preparation method and application, comprising hydrolyzed sodium hyaluronate, small molecule sodium chondroitin sulfate and water; wherein, by total mass percentage of raw materials, the hydrolyzed sodium hyaluronate is 1-20%, the small molecule sodium chondroitin sulfate is 0.05-5%, and the balance is water to 100%. The composition can promote skin cells to express AQP3, type I collagen and type III collagen, and promote skin cells to express type IV collagen, type VII collagen and type XVII collagen, etc., the main ECM of the basement membrane, so that the composition has both moisturizing and firming effects.

Description

Technical Field

[0001] The invention belongs to the technical field of cosmetics, and in particular relates to a basement membrane strip repairing composition and a preparation method and application thereof. Background Art

[0002] Human skin is composed of the epidermis, dermis and subcutaneous tissue, in which the epidermis and dermis form an interlocking structure at the junction, called the dermal-epidermal junction (DEJ), or the basement membrane zone. The interlocking structure of the basement membrane zone is conducive to the connection between the dermis and the epidermis, maintaining the firmness of the skin. The basement membrane zone of the skin contains rich extracellular matrix (ECM), mainly including: type IV collagen, type VII collagen, type XVII collagen, laminin, etc. Promoting the expression of these proteins is conducive to maintaining the normal function of the basement membrane zone. With age, the ability of skin cells to express various ECMs weakens, and the interlocking structure of the basement membrane zone of the skin gradually degenerates, causing the skin to dry and loose, leading to skin aging. Therefore, the repair of the basement membrane zone of the skin is an important research direction for skin anti-aging.

[0003] Sodium hyaluronate and sodium chondroitin sulfate are both important glycosaminoglycans in human skin, and are the main components of ECM, playing a major role in moisturizing the skin. Research reports indicate that sodium hyaluronate and sodium chondroitin sulfate can provide moisturizing and firming effects to the skin. For example, Chinese patent CN 202410135718.5 combines sodium heparin, sodium chondroitin sulfate, acetylated sodium hyaluronate, sodium hyaluronate and acetyl hexapeptide-8 to obtain an anti-wrinkle firming moisturizing composition.

[0004] However, conventional sodium hyaluronate and sodium chondroitin sulfate are both large molecular substances, which only play a moisturizing role in external skin care products and are difficult to be absorbed through the skin. Summary of the invention

[0005] The purpose of this section is to summarize some aspects of embodiments of the present invention and briefly introduce some preferred embodiments. Some simplifications or omissions may be made in this section and the specification abstract and the invention title of this application to avoid blurring the purpose of this section, the specification abstract and the invention title, and such simplifications or omissions cannot be used to limit the scope of the present invention.

[0006] In view of the above problems and / or the problems existing in the prior art, the present invention is proposed.

[0007] Therefore, the purpose of the present invention is to overcome the deficiencies in the prior art and provide a basement membrane repair composition.

[0008] In order to solve the above technical problems, the present invention provides the following technical solutions: a basement membrane zone repair composition, comprising hydrolyzed sodium hyaluronate, small molecule chondroitin sodium sulfate and water;

[0009] Wherein, based on the total mass percentage of the raw materials, the hydrolyzed sodium hyaluronate is 1-20%, the small molecule chondroitin sodium sulfate is 0.05-5%, and the balance is water to make up to 100%.

[0010] As a preferred embodiment of the composition of the present invention, the mass ratio of the hydrolyzed sodium hyaluronate to the small molecule chondroitin sodium sulfate is 1:5 to 20:1.

[0011] As a preferred embodiment of the composition of the present invention, the relative molecular weight of the hydrolyzed sodium hyaluronate is less than 50,000 Da.

[0012] As a preferred embodiment of the composition of the present invention, the relative molecular weight of the small molecule chondroitin sodium sulfate is less than 10000Da.

[0013] As a preferred embodiment of the composition of the present invention, the mass ratio of the hydrolyzed sodium hyaluronate to the small molecule chondroitin sodium sulfate is 1:5 to 5:1.

[0014] As a preferred embodiment of the composition of the present invention, the mass ratio of the hydrolyzed sodium hyaluronate to the small molecule chondroitin sodium sulfate is 1:1.

[0015] As a preferred embodiment of the composition of the present invention, the relative molecular weight of the hydrolyzed sodium hyaluronate is in the range of 400-3000Da.

[0016] As a preferred embodiment of the composition of the present invention, the relative molecular weight of the small molecule chondroitin sodium sulfate is in the range of 500 to 5000 Da.

[0017] Another object of the present invention is to overcome the deficiencies in the prior art and provide a method for preparing the composition, comprising weighing hydrolyzed sodium hyaluronate and small molecule chondroitin sodium sulfate, adding water and stirring until completely dissolved, filtering with a 0.45 μm water-based microporous filter membrane, filling into a sample bottle, and storing at 4° C. to obtain the composition.

[0018] Another object of the present invention is to overcome the deficiencies in the prior art and provide a basement membrane strip repair composition for use in preparing cosmetics with moisturizing and firming effects.

[0019] Beneficial effects of the present invention:

[0020] The invention provides a basement membrane zone repairing composition. By adopting a combination of hydrolyzed sodium hyaluronate and small molecule sodium chondroitin sulfate, a cosmetic raw material with the effect of promoting the repair of the basement membrane zone of the skin is obtained. The composition can promote skin cells to express AQP3, type I collagen and type III collagen, and promote skin cells to express type IV collagen, type VII collagen and type XVII collagen, etc., which are the main ECMs of the basement membrane zone, so that the composition has both moisturizing and firming effects. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the following briefly introduces the drawings required for describing the embodiments. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without creative labor. Among them:

[0022] Figure 1 This is a diagram showing the promoting effect of hydrolyzed sodium hyaluronate with different molecular weights on the expression of AQP3 in fibroblasts in the examples of the present invention.

[0023] Figure 2 This is a diagram showing the promoting effect of chondroitin sodium sulfate with different molecular weights on the expression of AQP3 in fibroblasts in the examples of the present invention.

[0024] Figure 3 This is a graph showing the effects of different mass ratios of the hydrolyzed sodium hyaluronate and small molecule chondroitin sodium sulfate compositions on the expression of AQP3 in fibroblasts in an embodiment of the present invention.

[0025] Figure 4 This is a diagram showing the promoting effect of the composition in the examples of the present invention on the expression of type I collagen and type III collagen.

[0026] Figure 5 The figure is a graph showing the promoting effect of the composition in the embodiment of the present invention on the expression of type IV collagen, type VII collagen and type XVII collagen.

[0027] Figure 6 This is a test chart of the water content of the human stratum corneum and the transepidermal water loss in the embodiment of the present invention.

[0028] Figure 7 This is a test chart of the amount of collagen in human skin in an embodiment of the present invention.

[0029] Figure 8 This is a test diagram of human skin dermis density and skin elasticity in an embodiment of the present invention. DETAILED DESCRIPTION

[0030] In order to make the above-mentioned objects, features and advantages of the present invention more obvious and easy to understand, the specific implementation methods of the present invention are described in detail below in conjunction with the embodiments of the specification.

[0031] In the following description, many specific details are set forth to facilitate a full understanding of the present invention, but the present invention may also be implemented in other ways different from those described herein, and those skilled in the art may make similar generalizations without violating the connotation of the present invention. Therefore, the present invention is not limited to the specific embodiments disclosed below.

[0032] Secondly, the term "one embodiment" or "embodiment" as used herein refers to a specific feature, structure, or characteristic that may be included in at least one implementation of the present invention. The term "in one embodiment" that appears in different places in this specification does not necessarily refer to the same embodiment, nor does it refer to a separate or selective embodiment that is mutually exclusive with other embodiments.

[0033] Example 1

[0034] Screening process of hydrolyzed sodium hyaluronate with different molecular weights:

[0035] Hydrolyzed sodium hyaluronate of 1294Da, 7266Da and 37358Da were selected for testing on promoting the expression of AQP3 in fibroblasts.

[0036] The determination method is:

[0037] Fibroblasts were grown in complete medium (high-glucose DMEM medium containing 10% FBS and 1% double antibody);

[0038] The cells were digested with trypsin and 2 × 10 5 Each well was inoculated into a 6-well plate, 2 mL per well, and cultured in a 37°C, 5% CO2 incubator for 24 h;

[0039] The culture medium was aspirated and the residual culture medium was washed with PBS, and 2 mL of SDS (sodium dodecyl sulfate, dissolved in PBS) with a concentration of 25 μg / mL was added to the rest, and incubated in the incubator for 4 hours. The culture medium was aspirated and the residual culture medium was washed with PBS, 2 mL of DMEM was added to the blank group, and 2 mL of hydrolyzed sodium hyaluronate with different molecular weights prepared in DMEM was added to the rest, and 3 parallels were added to each group, and incubated in the incubator. After 24 hours, the sample was aspirated, washed twice with PBS, and 200 μL of lysis buffer was added for lysis, sampling, centrifugation, and the supernatant was taken for use.

[0040] Refer to Elisa Kit for testing.

[0041] From above Figure 1It can be seen that at the same concentration, the ability of hydrolyzed sodium hyaluronate with a molecular weight of 1294Da to promote fibroblasts to produce AQP3 is better than that of hydrolyzed sodium hyaluronate with molecular weights of 7266Da and 37358Da, which is beneficial to deep moisturizing of the skin. Therefore, hydrolyzed sodium hyaluronate with a molecular weight of 1294Da was selected for subsequent experiments.

[0042] Example 2

[0043] Screening process of chondroitin sodium sulfate with different molecular weights:

[0044] Chondroitin sodium sulfate of 1410Da, 4048Da and 8296Da were selected for the determination of AQP3 content.

[0045] The AQP3 content detection method is the same as in Example 1.

[0046] Depend on Figure 2 It can be seen that at the same concentration, the ability of chondroitin sodium sulfate with a molecular weight of 4048Da to promote fibroblasts to produce AQP3 is better than that of chondroitin sodium sulfate with molecular weights of 1410Da and 8296Da, so chondroitin sodium sulfate with a molecular weight of 4048Da was selected for subsequent experiments.

[0047] Example 3

[0048] Screening process of the ratio of hydrolyzed sodium hyaluronate and small molecule chondroitin sodium sulfate:

[0049] A moisturizing composition is prepared by compounding the hydrolyzed sodium hyaluronate and small molecule sodium chondroitin sulfate obtained in Examples 1 and 2 in different mass ratios, wherein the compounding ratios are 5:1, 2:1, 1:1, 1:2, and 1:5, respectively.

[0050] Its preparation method:

[0051] Weigh hydrolyzed sodium hyaluronate and small molecule sodium chondroitin sulfate according to different mass ratios, add water and stir until completely dissolved, filter with a 0.45 μm water-based microporous filter membrane, fill into sample bottles, and store at 4°C.

[0052] The AQP3 content detection method is the same as in Example 1.

[0053] Depend on Figure 3 It can be found that when the mass ratio of hydrolyzed sodium hyaluronate with a molecular weight of 1294Da and sodium chondroitin sulfate with a molecular weight of 4048Da is 1:1, its ability to promote fibroblasts to produce AQP3 is better than other compound ratios, and has a significant moisturizing effect;

[0054] Therefore, the mass ratio of hydrolyzed sodium hyaluronate to small molecule sodium chondroitin sulfate was selected as 1:1 for subsequent experiments.

[0055] Example 4

[0056] The promoting effect of the composition on the expression of type I collagen and type III collagen by fibroblasts:

[0057] The specific method is as follows:

[0058] Fibroblasts were grown in complete medium (high-glucose DMEM medium containing 10% FBS and 1% double antibody);

[0059] Trypsinized cells were collected at a rate of 1 × 10 5 Each well was inoculated into a 96-well plate, 100 μL per well, and cultured in a 37°C, 5% CO2 incubator for 24 h;

[0060] The culture medium was aspirated and the residual culture medium was washed with PBS. Then 100 μL PBS was added and the UVA intensity was 10 J / cm 2 After irradiation, 100 μL of sample and DMEM were added and incubated in an incubator. After 24 hours, the upper culture medium was taken, centrifuged at 13000 rpm for 5 minutes, and the supernatant was taken to measure the content of type I collagen and type III collagen using Elisa Kit.

[0061] Depend on Figure 4 It can be obtained that the moisturizing composition promotes the expression of type I collagen and type III collagen within the concentration range of 0.05%-0.5%, and has a firming and anti-aging effect.

[0062] Example 5

[0063] The promoting effect of the composition on the expression of type IV collagen, type VII collagen and type XVII collagen:

[0064] The specific method is as follows:

[0065] Fibroblasts were grown in complete medium (high-glucose DMEM medium containing 10% FBS and 1% double antibody). Cells were trypsinized and 1×10 5 Each well was inoculated with 100 μL of cells per well in a 96-well plate and cultured in a 37°C, 5% CO2 incubator for 24 h. The culture medium was aspirated and the residual culture medium was washed with PBS. Then 100 μL of PBS was added and the UVA intensity was 10 J / cm 2 After irradiation, 100 μL of sample and DMEM were added and incubated in an incubator. After 24 hours, the supernatant was aspirated, the cells were lysed with lysis solution, the lysate was collected, centrifuged at 13000 rpm for 5 min, the supernatant was taken, and the content of type IV collagen and type VII collagen was determined using Elisa Kit.

[0066] Keratinocytes were grown in complete medium (high-glucose DMEM medium containing 10% FBS and 1% double-antibody). Cells were trypsinized and cultured at 1×10 5 Each well was inoculated with 100 μL of cells per well in a 96-well plate and cultured in a 37°C, 5% CO2 incubator for 24 h. The culture medium was aspirated and the residual culture medium was washed with PBS. Then 100 μL of PBS was added and the UVB intensity was 1 J / cm 2 After irradiation, 100 μL of sample and DMEM were added and incubated in an incubator. After 24 hours, the supernatant was aspirated, the cells were lysed with lysis solution, the lysate was collected, centrifuged at 13000 rpm for 5 min, the supernatant was taken, and the content of type ⅩⅦ collagen was determined using Elisa Kit.

[0067] Depend on Figure 5 The results show that the moisturizing composition promotes the expression of type IV collagen, type VII collagen and type XVII collagen within a concentration range of 0.05%-0.5%, and has the effect of repairing the basement membrane zone.

[0068] Example 6

[0069] Human testing uses samples prepared with 2% of the composition to verify efficacy

[0070] Examples of cosmetic formulations with moisturizing compositions:

[0071] The cream formula is as follows:

[0072] Raw material name Addition amount / % Remark water 74.5 Composition 2.0 The blank control was replaced with water Hydrogenated Lecithin 3.0 Polyglyceryl-10 Pentastearate 5.0 Squalane 10.0 Cetearyl Alcohol 3.0 Cyclopentasiloxane 2.0 Phenoxyethanol 0.45 Ethylhexylglycerin 0.05

[0073] Example 7

[0074] Human efficacy - skin water content and transepidermal water loss:

[0075] 24 hours in advance, 10 volunteers were subjected to skin barrier damage on the front of their arms with a 5% SDS aqueous solution, and then the skin moisture content and transepidermal water loss rate of the damaged area were tested. Based on this, blank formula (water was used instead of the moisturizing composition) and formula containing 2% moisturizing composition were applied to different areas, and the skin moisture content and transepidermal water loss rate of the applied area were tested before damage, 30 minutes after damage, 1 day, 3 days and 7 days.

[0076] Depend on Figure 6 It can be found that after applying a cream containing 2% of the moisturizing composition, the water content of keratin increases with the passage of time, the transepidermal water loss value of the skin is lower than that before application, and the repair effect is proportional to time, which proves that it has the effect of moisturizing and repairing the skin barrier function.

[0077] Example 8

[0078] Human body effect - skin collagen content:

[0079] 16 volunteers were selected to conduct a test experiment on the skin collagen content on their faces to investigate the promoting effect of moisturizing composition 1 on the production of facial skin collagen. The initial value of facial skin collagen was recorded before the test, and then the volunteers were required to apply a blank formula on one side of the face and a formula containing 2% moisturizing composition on the other side once in the morning and evening every day. The facial skin collagen content was tested after 7, 14, and 28 days respectively.

[0080] Depend on Figure 7 It can be seen that after applying the cream containing 2% of the moisturizing composition, the content of skin collagen gradually increases, which complements the results of Examples 4 and 5 and has the ability to promote skin collagen synthesis.

[0081] Example 9

[0082] Human body effects - skin dermal density and skin elasticity:

[0083] 16 volunteers were selected to conduct a test experiment on the dermal density and skin elasticity of the face to investigate the effect of the moisturizing composition on improving the dermal density and skin elasticity of the face skin. The initial values ​​of the dermal density and skin elasticity of the face skin were recorded before the test, and then the volunteers were required to apply a blank formula on one side of the face and a formula containing 2% of the moisturizing composition on the other side once a day in the morning and evening. The dermal density and skin elasticity of the face skin were tested after 7, 14, and 28 days respectively.

[0084] Depend on Figure 8 It is known that after applying a cream containing 2% of the wet composition, the dermal density and elasticity of the skin are significantly improved, and as the use time goes by, the effect becomes more significant, and it has the effect of tightening the skin.

[0085] It should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention rather than to limit it. Although the present invention has been described in detail with reference to the preferred embodiments, those skilled in the art should understand that the technical solutions of the present invention can be modified or replaced by equivalents without departing from the spirit and scope of the technical solutions of the present invention, which should all be included in the scope of the present invention.

Claims

1. A basement membrane repair composition, characterized in that: Including hydrolyzed sodium hyaluronate, small molecule chondroitin sodium sulfate and water; Wherein, based on the total mass percentage of the raw materials, the hydrolyzed sodium hyaluronate is 1-20%, the small molecule chondroitin sodium sulfate is 0.05-5%, and the balance is water to make up to 100%.

2. The composition according to claim 1, characterized in that: The mass ratio of the hydrolyzed sodium hyaluronate to the small molecule sodium chondroitin sulfate is 1:5 to 20:

1.

3. The composition according to claim 1 or 2, characterized in that: The relative molecular weight of the hydrolyzed sodium hyaluronate is lower than 50,000 Da.

4. The composition according to claim 3, characterized in that: The relative molecular weight of the small molecule chondroitin sodium sulfate is lower than 10000Da.

5. The composition according to claim 4, characterized in that: The mass ratio of the hydrolyzed sodium hyaluronate to the small molecule sodium chondroitin sulfate is 1:5 to 5:

1.

6. The composition according to claim 5, characterized in that: The mass ratio of the hydrolyzed sodium hyaluronate to the small molecule sodium chondroitin sulfate is 1:

1.

7. The composition according to claim 6, characterized in that: The relative molecular weight of the hydrolyzed sodium hyaluronate is in the range of 400-3000Da.

8. The composition according to claim 7, characterized in that: The relative molecular weight of the small molecule chondroitin sodium sulfate is in the range of 500 to 5000 Da.

9. The method for preparing the composition according to any one of claims 1 to 8, characterized in that: include, The hydrolyzed sodium hyaluronate and small molecule sodium chondroitin sulfate were weighed, water was added and stirred until completely dissolved, the mixture was filtered through a 0.45 μm water system microporous filter membrane, and then filled into a sample bottle and stored at 4° C. to obtain the composition.

10. Use of the composition according to any one of claims 1 to 8 in the preparation of cosmetics with moisturizing and firming effects.

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