Preparation and application of skin extracellular matrix containing human soluble collagen

Abstract

The invention discloses preparation and application of a skin extracellular matrix containing human soluble collagen. The skin extracellular matrix containing the human soluble collagen is prepared from the following components: 1 to 5 percent of human soluble collagen, 0.5 to 1.5 percent of tremella extract, 0.1 to 0.8 percent of hyaluronic acid, 2 to 6 percent of dipotassium glycyrrhizinate, golden orange and chrysanthemum water, 0.1 to 1.0 percent of arbutin, vitamin C, 0.13 to 1.5 percent of trehalose, 1 to 6 percent of hydroxypropyl tetrahydropyrantriol, 0.1 to 1.0 percent of scutellaria baicalensis root extract, 2 to 8 percent of butanediol and the balance of water. The skin extracellular matrix containing the human soluble collagen has remarkable moisturizing, repairing and anti-oxidation effects, and is suitable for postoperative skin repairing and daily nursing. The soluble collagen and the hyaluronic acid provide deep moisturizing, the tremella fuciformis extract and the golden orange and chrysanthemum water have anti-inflammatory and soothing effects, and the hydroxypropyl tetrahydropyrantriol and the scutellaria baicalensis root extract are beneficial to enhancing the skin barrier function. In addition, the preparation process of the product can be used for industrial large-scale production.

Description

Technical Field

[0001] The invention belongs to the technical field of bioengineering, and relates to the preparation and application of a skin extracellular matrix containing human soluble collagen. Background Art

[0002] The skin is the largest organ in the human body and plays an important role in protecting the skin from external stimuli and infections. The skin usually undergoes a recovery period after surgery, during which the skin barrier function is weakened and it is susceptible to external stimuli and infections. Therefore, the research and development of skin extracellular matrix containing human collagen for postoperative skin repair is particularly important.

[0003] Many skin care products on the market currently focus on moisturizing and anti-aging, but there are relatively few products that focus on postoperative skin repair. These products often take a long time to take effect, the repair effect is not obvious, and they cannot meet the special needs of postoperative skin.

[0004] In order to address the above problems, the patent proposes a skin extracellular matrix containing human collagen, whose ingredients include collagen, sodium hyaluronate, arbutin, tremella extract, etc. These ingredients can work synergistically to provide good moisturizing and anti-inflammatory soothing effects, and promote the recovery of the skin barrier.

[0005] The innovation of this kind of skin extracellular matrix containing human collagen lies in the scientific ratio of its ingredients and preparation method, which can significantly improve the skin's repair ability and meet the special needs of postoperative skin. Summary of the invention

[0006] The purpose of the present invention is to address the above-mentioned deficiencies of the prior art. The present invention first provides a skin extracellular matrix containing human soluble collagen, which comprises the following components: human soluble collagen, plant extracts, hyaluronic acid, dipotassium glycyrrhizinate, golden chamomile flower water, arbutin, antioxidants, trehalose, hydroxypropyl tetrahydropyrantriol, butylene glycol and water.

[0007] In certain embodiments, the antioxidant comprises vitamin C.

[0008] In certain embodiments, the plant extract includes Tremella fuciformis extract and Scutellaria baicalensis root extract.

[0009] In certain embodiments, it includes the following components: human soluble collagen, Tremella fuciformis extract, hyaluronic acid, dipotassium glycyrrhizinate, golden chamomile flower water, arbutin, vitamin C, trehalose, hydroxypropyl tetrahydropyrantriol, scutellaria baicalensis root extract, butylene glycol and water.

[0010] In certain embodiments, it includes the following components: 1-5% human soluble collagen, 0.5-1.5% Tremella fuciformis extract, 0.1-0.8% hyaluronic acid, dipotassium glycyrrhizinate, 2-6% golden chamomile flower water, 0.1-1.0% arbutin, vitamin C, 0.13-1.5% trehalose, 1-6% hydroxypropyl tetrahydropyrantriol, 0.1-1.0% scutellaria root extract, 2-8% butylene glycol, and the balance is water.

[0011] The present invention also provides a method for preparing the above-mentioned skin extracellular matrix containing human soluble collagen, characterized in that the method comprises the following steps:

[0012] (1) Dissolution stage: Add an appropriate amount of water to a clean container, gradually add butanediol, and stir evenly;

[0013] Add hyaluronic acid, trehalose and Tremella fuciformis extract in sequence and continue stirring until completely dissolved.

[0014] (2) Mixing stage: In another container, add human soluble collagen, slowly add the aqueous solution, and stir evenly;

[0015] Add dipotassium glycyrrhizinate, golden chamomile flower water, arbutin, vitamin C, hydroxypropyl tetrahydropyrantriol and scutellaria root extract, and continue stirring until smooth;

[0016] Combination stage: Combine the solutions in the two containers and stir slowly to ensure that all ingredients are fully mixed; adjust the pH value to ensure the stability and skin adaptability of the product.

[0017] In some embodiments, it further comprises the following steps:

[0018] (4) Homogenization: Use a homogenizer to homogenize the mixture to ensure uniform distribution of ingredients and improve the texture and absorption effect of the product;

[0019] (5) Filling and packaging: The homogenized human collagen-containing skin extracellular matrix is ​​filled into a clean container and sealed to avoid contact with light and air to ensure the stability of the product.

[0020] In certain embodiments, the pH value is adjusted to a pH value of 5.5-6.5.

[0021] Finally, the present invention provides the use of the above-mentioned skin extracellular matrix containing human soluble collagen in skin care, wherein the use is for non-therapeutic purposes.

[0022] In certain embodiments, the application is one or more of supporting skin, moisturizing skin, skin allergy prevention, skin wound recovery, scar reduction, and cosmetic post-operative care.

[0023] Compared with the prior art, the present invention has the following advantages and beneficial effects:

[0024] In the skin extracellular matrix containing human soluble collagen described in the present invention, human soluble collagen can promote the regeneration and repair of skin cells, accelerate the recovery process of damaged skin, reduce the formation of scars, improve the healing quality of damaged skin, interact with elastic fibers in the skin, improve the elasticity of the skin, and also have antioxidant ability, reducing the damage of free radicals to the skin; Tremella extract is a polysaccharide compound, known as "plant hyaluronic acid", which not only has a strong water-locking effect, but also has anti-inflammatory and soothing effects, delays skin aging, and is safe, gentle and non-irritating to human skin; hyaluronic acid, as a commonly used moisturizer in skin care products, can more effectively increase the water content in the skin, and at the same time cooperate with glycyrrhizic acid dimethyl to inhibit the degradation of hyaluronic acid to prevent oligopeptides from irritating the skin; golden chamomile flower water and scutellaria root extract contain flavonoids The present invention discloses a substance which is rich in antioxidant ingredients and can neutralize free radicals and reduce the damage of oxidative stress to the skin. In addition, the scutellaria root extract can neutralize histamine and reduce skin allergic reactions, and also has whitening, freckle removal and anti-inflammatory effects. Arbutin is a commonly used whitening conditioning agent, which inhibits the activity of tyrosinase and prevents the formation of melanin. Vitamin C is added as an antioxidant to maintain the activity of arbutin. Trehalose is a natural sugar with hydrophilicity and forms a hydration film on the surface of the skin. It complements and enhances the synergy with hyaluronic acid and also has the effects of promoting cell growth and protecting the biological activity and stabilization of other macromolecules. Hydroxypropyl tetrahydropyrantriol is a xylose derivative and is widely used in skin care products. It promotes the formation of collagen by inducing the biosynthesis of glycosaminoglycans (GAGs) in the superficial epidermis of the skin, enhances the elasticity and firmness of the skin, and also has moisturizing and anti-aging effects.

[0025] The skin extracellular matrix containing human soluble collagen of the present invention combines recombinant human type III collagen with natural extracts or product raw materials, and the components work together to enhance the effect. The product not only has excellent moisturizing properties, but also can play an anti-allergic, repairing and soothing role on the skin during use, effectively responding to allergenic stimuli in daily life, and providing soothing care. In addition, the product has a gentle and moisturizing texture, is easily absorbed by the skin, and has an excellent user experience. In addition, the skin extracellular matrix containing human soluble collagen of the present invention is particularly suitable for cosmetic postoperative care. Since the skin barrier after cosmetic surgery is relatively fragile and easily stimulated by the outside world, the human soluble collagen and natural extracts in the skin extracellular matrix containing human soluble collagen can effectively promote skin repair, reduce redness and discomfort, and help the skin return to a healthy state. Its mild formula ensures comfort during use, making it an ideal choice for cosmetic postoperative care. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 Soluble collagen 1.

[0027] Figure 2 Soluble collagen 2;

[0028] Figure 3 Soluble collagen 3;

[0029] Figure 4 Tremella fuciformis extract;

[0030] Figure 5 Scutellaria baicalensis extract;

[0031] Figure 6 Hyaluronic acid;

[0032] Figure 7 Elastase activity inhibition test diagram; 0.01<p<0.05 significant "*"; 0.001<p<0.01 significant "**"; 0.0001<p<0.001 significant "***"; p<0.0001 significant "****";

[0033] Figure 8 Figure 2. Hyaluronidase activity inhibition experiment. DETAILED DESCRIPTION

[0034] In order to make the purpose, technical scheme and advantages of the present invention clearer, the specific embodiments of the present invention are further described in detail below in conjunction with specific examples and drawings. If no specific technology or conditions are specified in the examples, the technology or conditions described in the literature in this field or the product instructions are used. If the manufacturer of the reagents or instruments is not specified, they are all conventional products that can be obtained commercially.

[0035] Example 1

[0036] S1·Preparation of human soluble collagen 1

[0037] The sequence of the recombinant type III collagen used in this example is

[0038] GLAGYPGPAGPPGPPGPPGTSGHPGSPGSPGYQGPPGEPGQAGPSGPPGPPG

[0039] AIGPSGPAGKDGESGRPGAPGERGRPGLPGAAGARGNDGARGSDGQPGPP

[0040] GPPGTAGFPGSPGAKGEVGPAGPSPGSNGAPGQRGHAGAQGPPGPPGINGSP

[0041] GGKGEMGPAGIPGAPGLMGARGPPGPAGANGAPGLRGGAGEPGKNGAKG

[0042] KNGETGPQGPPGPTGPGGDKGDTGPPGPQGLQGLPGTGGPPGENGKPGEP

[0043] GPKGDAGAPGAPGGKGDAGAPGERGPPGLAGAPGLRGVPGKDGPRGPTG

[0044] PIGPPGPAGQPGDKGEGGAPLPGIAGPRGSPGERGETGPPGPAGFPGAPGQ

[0045] NGEPGNPGPPGPSGSPGKDGPPGPAGNTGAPGSPGVSGPKGDAGQPGEKG

[0046] SPGAQGPPGAPGPLGIAGITGMPGPRGSPGPQGVKGESGKPGANGLSGERG

[0047] PPGPQGLPGLAGTAGEPGRDGNPGSDGLPGRDGSPGDKGETGERGAAGIKGHRGFPGNPGAPGSPGPAGQQGAIGSPGPA (SEQ ID No. 1).

[0048] The preparation process is completed by referring to the scheme of CN118184767A.

[0049] S2 Dissolution stage:

[0050] Add an appropriate amount of water to a clean container, gradually add butanediol and stir evenly.

[0051] Add hyaluronic acid, trehalose and Tremella fuciformis extract in sequence and continue stirring until completely dissolved.

[0052] S3 Mixing stage:

[0053] In another container, add human soluble collagen, slowly add the aqueous solution, and stir evenly.

[0054] Add dipotassium glycyrrhizinate, golden chamomile flower water, arbutin, vitamin C, hydroxypropyl tetrahydropyrantriol and scutellaria baicalensis root extract and continue stirring until smooth.

[0055] S4·Merge stage:

[0056] Combine the solutions in both containers and stir slowly to ensure all ingredients are thoroughly mixed.

[0057] Adjust the pH value to 5.5-6.5 to ensure product stability and skin adaptability.

[0058] S5 Homogenization:

[0059] Use a homogenizer to homogenize the mixture to ensure that the ingredients are evenly distributed, improving the texture and absorption effect of the product.

[0060] S6 Filling and packaging:

[0061] The homogenized human collagen-containing skin extracellular matrix is ​​filled into a clean container and sealed to avoid contact with light and air to ensure the stability of the product.

[0062] Example 2

[0063] Preparation of human soluble collagen 2

[0064] The sequence of the recombinant type III collagen used in this example is

[0065] GLMGARGPPGPAGANGAPGLRGGAGEPGKNGAKGEPGPRGERGEAGIPGVPGAKGEDGKDGSPGEPGANGLPGAAGERGAPGFRGPAGPNGIPGEKGPAGERGAPGPAGPRGAAGEPGRDGVPGGPGMRGMPGLMGARGPPGPAGANGAPGLRGGAGEPGKNGAKGEPGPRGERGEAGIPGVPGAKGEDGKDGSPGEPGA NGLPGAAGERGAPGFRGPAGPNGIPGEKGPAGERGAPGPAGPRGAAGEPGRDGVPGGPGMRGMPGLMGARGPPGPAGANGAPGLRGGAGEPGKNGAKGEPGPRGERGEAGIPGVPGAKGEDGKDGSPGEPGANGLPGAAGERGAPGFRGPAGPNGIPGEKGPAGERGAPGPAGPRGAAGEPGRDGVPGGPGMRGMP(SEQ ID No.2).

[0066] The other steps are the same as in Example 1.

[0067] Example 3

[0068] Preparation of human soluble collagen 3

[0069] The sequence of the recombinant type III collagen used in this example is

[0070] GPPGIKGPAGIPGFPGMKGHRGFDGRNGEKGETGAPGLKGENGLPGENGAPGPMGPRGAPGERGRPGLPGAAGARGNDGARGSDGQPGPPGPPGTAGFPGSPGAKGEVGPAGSPGSNGAPGQRGEPGPQGHAGAQGPPGPPGINGPLGIAGITGA RGLAGPPGMPGPRGSPGPQGVKGESGKPGANGLSGERGPPGPQGLPGLAGTAGEPGRDGNPGSDGLPGRDGSPGKGDRGENGSPGAPGAPGHPGPPGPVGPAGKSGDRGESGPAGPAGAPGPAGSRGAPGPQGPRGDKGETGERGAAGIK(SEQ ID No. 3).

[0071] The other steps are the same as in Example 1.

[0072] Comparative Example 1

[0073] No recombinant collagen was added, and other steps were the same as in Example 1.

[0074] Comparative Example 2

[0075] No hyaluronic acid was added, soluble collagen 3 was added, and the other steps were the same as in Example 1;

[0076] Comparative Example 3

[0077] No Scutellaria baicalensis extract was added, soluble collagen 3 was added, and the other steps were the same as in Example 1;

[0078] Comparative Example 4

[0079] Without adding Tremella fuciformis extract, soluble collagen 3 was added, and the other steps were the same as those in Example 1;

[0080] Experiments of Examples 1-3 and Comparative Example 1:

[0081] Experiment 1: Cellular activity test of active substances contained

[0082] RAW264.7 cells were cultured in cell culture medium supplemented with 10% (v / v) fetal bovine serum (FBS, Cytivia) in an atmosphere of 5% CO2 / 95% at 37°C. To evaluate cytotoxicity, cells (8 × 10 3Cells / well) were inoculated into 96-well plates and cultured for 24 hours. Subsequently, different concentrations of recombinant humanized soluble collagen, Tremella extract, hyaluronic acid, and Scutellaria extract were added to each well, and the cells were cultured for another 72 hours. After removing the culture medium, 100 μL 10% CCK-8 / DMEM (v / v) was added to each well and cultured for 2 hours. The absorbance of each well at a wavelength of 450 nanometers was measured using a spectrophotometer (ThermoFisher Scientific). Cell viability (%) and IC50 values ​​were calculated using GraphPad Prism5.01 software.

[0083] See also Figure 1 Table 1: Recombinant humanized soluble collagen, Tremella fuciformis extract, Scutellaria baicalensis extract, and hyaluronic acid have good cell activity under certain concentration conditions. Cell absorption can be promoted within a certain concentration range.

[0084] Table 1 Cell viability of each component

[0085] Sample Cell survival rate (%) Soluble collagen 1 97.9 Soluble collagen 2 98.2 Soluble collagen 3 98.6 Scutellaria baicalensis extract 97.9 Hyaluronic acid 97.3 Tremella fuciformis fruiting body extract 98.5

[0086] Experiment 2: Anti-inflammatory test of active substances contained

[0087] RAW264.7 cells were cultured in DMEM cell culture medium supplemented with 10% (v / v) fetal bovine serum (FBS, Cytivia) in an environment of 5% CO2 / 95% at 37°C. To evaluate the anti-inflammatory effect, cells (8 × 10 3 Cells / well) were inoculated into 96-well plates and cultured for 24 hours. Subsequently, the cells were divided into a control group, an LPS group, and groups treated with different concentrations of the test components. The control group was added with culture medium; 1 μg / ml of LPS was added to the LPS group; different concentrations of the test components were added to the treatment group, and 1 μg / ml of LPS was added at the same time. The treated cells were incubated at 37°C and 5% CO2 for 24 hours. The cell supernatant was collected, and the concentrations of inflammatory factors such as TNF-α and IL-6 were detected using an ELISA kit. The concentrations of inflammatory factors in each group were compared to evaluate the anti-inflammatory effect of the test components.

[0088] See also Figure 1-Figure 6 : Recombinant humanized soluble collagen, Tremella fuciformis extract, Scutellaria baicalensis extract, and hyaluronic acid all have significant anti-inflammatory effects under certain concentration conditions, which verifies the anti-inflammatory effect of the present invention.

[0089] Experiment 3, Example 1-3 Elastase Activity Inhibition Experiment

[0090] Dilute the sample with PBS N times (dilute to a relatively clear and fluid state; if retesting is required, adjust according to the test results) as a sample for later use. Group test: Add the sample, elastase solution, and positive control agent to the corresponding wells of the sample group, blank control group, and positive control group, as shown in the chart.

[0091] Test process: as listed in the table, after adding the elastase solution, incubate at 37°C for 10 minutes; add the remaining reagents in Table 1 to the corresponding wells of the test group, control group, and positive control group. Except for the blank group, add biotin-labeled antibodies and enzyme-labeled reagents to each well, incubate at 37°C for 60 minutes, and then wash the plate; add the color developing solution, add the stop solution after color development at 37°C for 15 minutes, and measure the OD value at 450nm with an enzyme reader. Calculate the elastase activity inhibition rate according to the formula shown below.

[0092]

[0093] Where:

[0094] C is the average absorbance value of the control group;

[0095] T is the average absorbance of the test group;

[0096] B is the average absorbance value of the blank group;

[0097] P is the average absorbance value of the positive control group.

[0098] Results: Under the test conditions, Figure 7 According to the analysis of Table 2, the inhibition rate of elastase in the examples of the present invention at 50% of the tested concentration is higher than 30%, which is significantly different from the data of the control group (p<0.0001). In addition, in the control group without recombinant type III collagen, the inhibition rate of elastase is 15%. This proves that the recombinant type III collagen in Examples 1 to 3 has an excellent inhibitory effect on elastase. According to the correlation between elastase and firming and anti-wrinkle, all examples have firming and anti-wrinkle effects, among which Example 3 has the most significant effect.

[0099] Table 2 Test liquid addition grouping

[0100] Group Elastase Buffer 50mg / mL ascorbic acid Test samples T-test group + - - + C-Control Group + + - - P-positive control group + - + - B-Blank group - - - -

[0101] Experiment 4, Example 1-3 Hyaluronidase Activity Inhibition Experiment

[0102] Sample treatment: Dilute the sample with PBS N times (N is initially set to 10, and then adjusted according to the test results) as a sample for standby use. Group test: Add the sample, hyaluronidase, and positive control agent to the corresponding wells of the sample group, blank control group, and positive control group, as shown in Table 2.

[0103] Test process: as listed in the table, after adding hyaluronidase solution, incubate at 37°C for 10 minutes; add the remaining reagents in Table 1 to the corresponding wells of the test group, control group, and positive control group. Except for the blank group, add biotin-labeled antibodies and enzyme-labeled reagents to each well, incubate at 37°C for 60 minutes, and then wash the plate; add color developing solution, add stop solution after color development at 37°C for 15 minutes, and measure the OD value at 450nm with an enzyme reader. Calculate the elastase activity inhibition rate according to the formula shown below.

[0104]

[0105] Where:

[0106] C is the average absorbance value of the control group;

[0107] T is the average absorbance of the test group;

[0108] B is the average absorbance value of the blank group;

[0109] P is the average absorbance value of the positive control group.

[0110] Results: Under the test conditions, Figure 8 According to the analysis of Table 3, the inhibition rate of the embodiments of the present invention on hyaluronidase at 50% test concentration is about 20%, which is significantly different from the data of the control group (p < 0.01), proving that it has an inhibitory effect on hyaluronidase. According to the correlation between hyaluronidase and soothing, the embodiments of the present invention have a soothing effect, and the embodiment 3 has the best effect.

[0111] Table 3 Test liquid addition grouping

[0112] Group Hyaluronidase Buffer 30mg / mL dipotassium glycyrrhizinate Test samples T-test group + - - + C-Control Group + + - - P-positive control group + - + - B-Blank group - - - -

[0113] Experiment 5, Hydroxyl Radical Scavenging Test

[0114] Dilute the sample with ultrapure water N times (dilute until it is clear and has good fluidity; if retesting is required, make adjustments based on the test results) and set it aside as a sample.

[0115] According to Table 4, 9 mmol / L FeSO4 and 9 mmol / L ethanol-salicylic acid were added to each solution, followed by an appropriate amount of sample, then an appropriate amount of ultrapure water, and finally 8.8 mmol / L hydrogen peroxide. After heating in a 37°C water bath for 15 minutes, the absorbance of the test object was measured at 510 nm.

[0116]

[0117] Where:

[0118] C is the average absorbance value of the control group;

[0119] T is the average absorbance of the test group;

[0120] C0 is the average absorbance value of the blank control group;

[0121] The results are shown in Table 4 and Figure 8 As shown: the active ingredients contained in Examples 1 to Comparative Examples 4 have good effects on scavenging hydroxyl radicals, among which the group containing soluble collagen is more effective in scavenging free radicals, especially Example 3 has more excellent antioxidant and anti-aging effects. The hydroxyl radical scavenging effect of Comparative Examples 2 to 4 is worse than that of Examples 1 to 3, but higher than that of Comparative Example 1, indicating that under the action of soluble collagen, the hydroxyl radical scavenging rate can be significantly improved, and under the effective mutual matching of other preferred ingredients, a synergistic enhancement effect can be achieved.

[0122] Table 4 Test liquid addition grouping

[0123]

[0124] The total reaction volume was 15 mL

[0125] Experiment 6, tyrosinase activity assay

[0126] 100 μL of extracellular matrix was added to the well plate inoculated with B16F10 cells, and pure water was used to replace the extracellular matrix as a blank sample. After 48 h of culture, trypsin was digested and about 10 7 cells, and then washed twice with PBS, centrifuged at 1000rpm for 5min, and the cell pellet was set aside. 1mL 0.5% sodium deoxycholate was added to the cell pellet and placed at 0℃ for 15min. After the time was up, 3mL 0.1% L-DOPA (prepared with 0.1M phosphate buffer with pH=6.8, the reaction mixture should be used within 2h after preparation) was added to the above mixture, the reaction temperature was 37℃, and the absorbance values ​​at 0min and 10min of the reaction were detected at a wavelength of 475nm to calculate the inhibition rate.

[0127] Results: See Table 5; The synthesis of melanin is affected by the activity of tyrosinase, which inhibits the production of melanin. By inhibiting the production and activity of tyrosinase, or interfering with the intermediates of melanin production, the production of melanin that produces pigment spots can be prevented. From the experimental results, it can be seen that the tyrosinase inhibition rate of the soluble collagen is higher, indicating that the soluble collagen has good whitening and spot-lightening effects, delays pigment deposition, the formation of spots, and has anti-aging effects. In addition, the tyrosinase inhibition rate of Comparative Examples 2 to 4 is significantly lower than that of Examples 1 to 3, but higher than that of Comparative Example 1, proving that under the action of soluble collagen, the formation of melanin can be more inhibited, and under the effective mutual matching of other preferred ingredients, a synergistic enhancement effect can be achieved.

[0128] Table 5 Free radical scavenging rate and tyrosinase activity results

[0129] Sample Hydroxyl free radical scavenging rate (%) Tyrosinase inhibition rate (%) Example 1 74.2 30.1 Example 2 76.9 31.2 Example 3 79.9 35.8 Comparative Example 1 40.6 11.6 Comparative Example 2 60.2 19.4 Comparative Example 3 63.3 18.7 Comparative Example 4 68.7 16.8

[0130] Experiment 7, Skin water loss and redness value detection of Examples 1-3

[0131] 31 subjects used the test products as required for 28 days and underwent skin moisture and redness tests. The subjects were aged 21 to 44 years old, and the number of subjects analyzed was 31.

[0132] The results are shown in Table 6. After continuous use of this product, the skin moisture loss of 31 subjects decreased, indicating that this product has a good effect on skin barrier moisture retention and stronger skin barrier tolerance. The moisture loss value 28 days after using the sample was significantly reduced compared with that before using the sample (****P<0.001). In addition, the skin redness of 31 subjects decreased after continuous use of this product, indicating that this product has a good repair effect on the skin barrier.

[0133] Table 6 Skin water loss through the skin and redness value detection

[0134] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. A skin extracellular matrix containing human soluble collagen, characterized in that: It includes the following ingredients: human soluble collagen, plant extract, hyaluronic acid, dipotassium glycyrrhizinate, golden chamomile flower water, arbutin, antioxidants, trehalose, hydroxypropyl tetrahydropyrantriol, butylene glycol and water.

2. The skin extracellular matrix containing human soluble collagen according to claim 1, characterized in that: The antioxidants include vitamin C.

3. The skin extracellular matrix containing human soluble collagen according to claim 1, characterized in that: The plant extracts include Tremella fuciformis extract and Scutellaria baicalensis root extract.

4. The skin extracellular matrix containing human soluble collagen according to claim 1, characterized in that: It includes the following components: human soluble collagen, tremella extract, hyaluronic acid, dipotassium glycyrrhizinate, golden chamomile flower water, arbutin, vitamin C, trehalose, hydroxypropyl tetrahydropyrantriol, scutellaria root extract, butylene glycol and water.

5. The skin extracellular matrix containing human soluble collagen according to claim 4, characterized in that: The invention comprises the following components: 1-5% human soluble collagen, 0.5-1.5% tremella extract, 0.1-0.8% hyaluronic acid, dipotassium glycyrrhizinate, 2-6% golden chamomile flower water, 0.1-1.0% arbutin, 0.13-1.5% vitamin C, trehalose, 1-6% hydroxypropyl tetrahydropyrantriol, 0.1-1.0% scutellaria root extract, 2-8% butylene glycol, and the balance is water.

6. A method for preparing a skin extracellular matrix containing human soluble collagen according to any one of claims 1 to 5, characterized in that: The method comprises the following steps: (1) Dissolution stage: Add an appropriate amount of water to a clean container, gradually add butanediol, and stir evenly; Add hyaluronic acid, trehalose and Tremella fuciformis extract in sequence and continue stirring until completely dissolved. (2) Mixing stage: In another container, add human soluble collagen, slowly add the aqueous solution, and stir evenly; Add dipotassium glycyrrhizinate, golden chamomile flower water, arbutin, vitamin C, hydroxypropyl tetrahydropyrantriol and scutellaria root extract, and continue stirring until smooth; Combination stage: Combine the solutions in the two containers and stir slowly to ensure that all ingredients are fully mixed; adjust the pH value to ensure the stability and skin adaptability of the product.

7. The method for preparing the skin extracellular matrix containing human soluble collagen according to claim 6, characterized in that: It also includes the following steps: (4) Homogenization: Use a homogenizer to homogenize the mixture to ensure uniform distribution of ingredients and improve the texture and absorption effect of the product; (5) Filling and packaging: The homogenized human collagen-containing skin extracellular matrix is ​​filled into a clean container and sealed to avoid contact with light and air to ensure the stability of the product.

8. The method for preparing the skin extracellular matrix containing human soluble collagen according to claim 6, characterized in that: The pH value is adjusted to 5.5-6.

5.

9. Use of the skin extracellular matrix containing human soluble collagen according to any one of claims 1 to 5 in skin care, wherein the use is for non-therapeutic purposes.

10. The use according to claim 9, characterized in that: The application is one or more of supporting skin, moisturizing skin, anti-allergic skin, skin wound recovery, reducing scars, and cosmetic post-operative care.

Images