A skin repair composition containing plant callus and preparation method thereof

Abstract

The present invention relates to a skin repair composition containing plant callus and a preparation method thereof, and belongs to the technical field of skin repair. A skin repair composition containing plant callus comprises the following components: fir callus extract, ergothioneine, conopeptide, squalane and sustained-release material; the preparation of the sustained-release material comprises: after mixing soybean lecithin, cholesterol and anhydrous ethanol, ultrasonic treatment, evaporation dispersion; after mixing hesperidin, composite antioxidant and liposome, ultrasonic vibration after heating, freeze drying, and obtain; the present invention adopts citral to react with chitosan oligosaccharide, increases stability and increases antibacterial performance, and ergothioneine and composite antioxidant have interaction force through conjugation effect, which can affect the configuration and antibacterial activity of composite antioxidant molecules, and cooperates with hesperidin, chitosan oligosaccharide and citral to significantly improve the antibacterial performance of the composition through synergistic effect.

Description

Technical Field

[0001] The invention belongs to the technical field of skin repair and relates to a skin repair composition containing plant callus. Background Art

[0002] Most cosmetic surgeries will cause a certain degree of trauma, which takes some time to heal. During the operation, external forces such as light and electricity can cause damage to the skin, making it temporarily sensitive. In addition, external stimulation can also cause the skin to release inflammatory factors, causing adverse reactions such as redness, swelling, and itching. If the skin is not effectively repaired after cosmetic surgery, it will accelerate skin aging. Therefore, in order to maintain skin health and provide a good recovery environment, antioxidant and antibacterial ingredients are essential. They are the top priority for skin repair, anti-aging, and anti-inflammation.

[0003] Hesperidin is the main active ingredient in the fruit of the genus Citrus in the Rutaceae family. Hesperidin is the glycosyl ligand of hesperidin, and its molecular formula is C 16 H 14 O6, its structure contains keto carbonyl, ether, methoxy and multiple phenolic hydroxyl groups, which give it a wide range of pharmacological effects. Hesperidin does not accumulate in any organ, is safe to use and has no obvious side effects. Early studies on the pharmacological effects of hesperidin mainly focused on antibacterial, anti-inflammatory, antioxidant, antiviral, lipid-regulating, immune-enhancing and anti-cancer effects.

[0004] As a natural compound, hesperidin has the advantages of abundant sources, high yield, low production cost, precise biological activity, and low toxicity and side effects, but it has disadvantages such as poor water solubility and low bioavailability. With the synthesis of hesperidin derivatives and the application of biotechnology pharmaceutical preparations, the disadvantages of hesperidin in terms of water solubility and bioavailability have been improved.

[0005] Chitosan is the product of N-deacetylation of chitin. The chemical structures of chitin, chitosan and cellulose are similar. Cellulose contains a hydroxyl group at the C2 position, while chitin and chitosan are replaced by acetylamino and amino groups at the C2 position, respectively. Chitin and chitosan have unique properties such as biodegradability, cell affinity and biological effects. In particular, chitosan containing free amino groups has become the only alkaline polysaccharide among natural polysaccharides. The molecular weight and deacetylation degree of chitosan have a significant effect on its antibacterial properties. Reducing the molecular weight can improve the antibacterial effect to a certain extent. However, although chitosan has certain bactericidal activity, its stability is weaker than that of conventional antibacterial materials, and its antibacterial activity is relatively low when used alone.

[0006] Citral is found in different plant sources. It is a monoterpene composed of two isomers, neral and geranial. In recent years, studies have shown that this compound exhibits a variety of biological activities, such as antibacterial, antifungal, antibiofilm, antiparasitic, antiproliferative, anti-inflammatory and antioxidant properties through in vitro and in vivo assays. Citral has a monoterpene structure and is easily chemically degraded by low pH, heat and oxygen. The low water solubility of citral leads to limitations in its application. It is also sensitive to volatilization under normal storage conditions, resulting in reduced bactericidal activity. Summary of the invention

[0007] The object of the present invention is to provide a skin repair composition containing plant callus and a preparation method thereof. The present invention adopts citral to react with chitosan oligosaccharide to increase stability and antibacterial performance at the same time, and ergothioneine and a composite antioxidant have an interactive force through a conjugated effect, which can affect the configuration and antibacterial activity of the composite antioxidant molecule, and hesperidin, chitosan oligosaccharide and citral are combined to significantly improve the antibacterial performance of the composition through a synergistic effect.

[0008] The purpose of the present invention can be achieved through the following technical solutions:

[0009] A skin repairing composition containing plant callus tissue comprises the following components in parts by weight:

[0010] 7-11 parts of fir callus extract, 2-3 parts of ergothioneine, 3-6 parts of conopeptide, 4-6 parts of squalane and 3.5-5.0 parts of sustained-release material; squalane not only helps to moisturize and repair the skin barrier, but also has good permeability and can enter the skin, replenish cell lipids and nourish the skin.

[0011] Further, the uptake process of ergothioneine requires the help of the transporter OCTN1, which exists in the keratinocytes and melanocytes of the skin and has a high affinity for ergothioneine. Therefore, adding ergothioneine to the drug helps other ingredients to be quickly accepted, obtained and utilized by skin tissue cells, thereby improving the efficacy of the composition of the present invention. It is worth noting that ergothioneine is different from water-soluble antioxidants (such as flavonoids, phenolic acids and polyphenols) in that it does not penetrate through the cell membrane. Hesperidin, citral and chitosan oligosaccharides of the present invention make up for this deficiency and enhance their antibacterial properties through synergistic effects.

[0012] In addition, ergothioneine has the characteristics of regulating intracellular redox reaction, participating in energy regulation, becomes the physiological protective agent of cell, and shows good safety and stability. The present invention, by adding ergothioneine, not only maintains the stability of other components, but also promotes the absorption of active ingredients by cells, realizes good synergistic effect, thereby effectively improving the effect of other components.

[0013] The preparation method of the sustained-release material comprises the following steps:

[0014] S1, mixing soybean lecithin, cholesterol and anhydrous ethanol, ultrasonically treating, evaporating and dispersing, to obtain liposomes;

[0015] S2, mixing hesperidin, composite antioxidant and liposome, heating until the liposome is hydrated, ultrasonically shaking, and freeze-drying to obtain a sustained-release material;

[0016] The preparation method of the composite antioxidant comprises the following steps:

[0017] 1) Chitosan, acetic acid and deionized water are stirred and mixed, and then hydrogen peroxide is added and stirred evenly for degradation treatment to obtain a chitosan oligosaccharide mixture;

[0018] 2) Under an inert atmosphere, citral and ethanol are stirred and mixed, and then ammonium cerium nitrate solution and chitosan oligosaccharide mixture are added in sequence and mixed, and after stirring for reaction, acetone is added for precipitation, and after standing, filtering, washing and drying, a composite antioxidant is obtained.

[0019] As a preferred technical solution of the present invention, in step S1, the mass ratio of soybean lecithin to cholesterol is 4.0-4.3:1.0-1.2; the conditions of the ultrasonic treatment are 100-120KHz frequency and 30-40min of ultrasound; the conditions of the evaporation dispersion are 50-60r / min speed and 30-35°C temperature of evaporation dispersion.

[0020] As a preferred technical solution of the present invention, in step S2, the mass ratio of hesperidin, composite antioxidant and liposome is 0.5-0.8:1.4-2.0:3.2-4.0; the conditions of the ultrasonic oscillation are 200-220r / min speed, 300-400MHz frequency, and treatment for 15-20min.

[0021] As a preferred technical solution of the present invention, in step 1), the mass ratio of chitosan, acetic acid, deionized water and hydrogen peroxide is 3-9:4-6:120:7-9; the molecular weight of chitosan is 120-150kDa; the conditions of the degradation treatment are stirring at a speed of 300-400r / min, an ultrasonic frequency of 65-80KHz, a temperature of 50-70°C, and degradation for 4-8h; the hydrogen peroxide is H2O2.

[0022] As a preferred technical solution of the present invention, in step 1), the molecular weight of chitosan oligosaccharide in the chitosan oligosaccharide mixture is 2000-2500.

[0023] As a preferred technical solution of the present invention, in step 2), the usage ratio of citral, ethanol, ammonium cerium nitrate solution and chitosan oligosaccharide mixture is 3.0-3.5g:22-25mL:0.5-0.8g:310-330g; the ethanol is anhydrous ethanol.

[0024] As a preferred technical solution of the present invention, in step 2), the inert atmosphere is a nitrogen atmosphere; the stirring reaction conditions are: after treating with an ultrasonic frequency of 30-40KHz for 5-8min, stirring at a speed of 200-300r / min, at a temperature of 35-40°C, and reacting for 4-6h.

[0025] As a preferred technical solution of the present invention, in step 2), the concentration of the ammonium cerium nitrate solution is 0.04-0.05 g / mL.

[0026] As a preferred technical solution of the present invention, in step 2), the standing time is 1-2 hours; the washing condition is washing with anhydrous ethanol until neutral; and the drying is freeze-drying.

[0027] The invention discloses a method for preparing a skin repairing composition containing plant callus tissue. The preparation method comprises the following steps: weighing each component by weight and mixing the components to obtain a skin repairing composition.

[0028] As a preferred technical solution of the present invention, the fir explants are selected from the branches of North American redwood plants after disinfection; the method for preparing the fir callus extract filtrate comprises the following steps:

[0029] S11, placing the Chinese fir explants in an agar medium to perform callus induction culture. This process needs to be carried out at a temperature of 20°C for 14 days to obtain callus;

[0030] S12, transfer the obtained callus tissue to MS medium, place it in a shaking incubator, and perform shaking culture at a speed of 100r / min. Similarly, the temperature of this process should be controlled at 20°C for 14 days to obtain proliferating callus tissue;

[0031] S13, inoculating the proliferated callus into a sterile cell bag for expansion culture. This process should be carried out at a temperature of 25°C for 24 days to obtain the Chinese fir callus culture;

[0032] S14, filtering the Chinese fir callus culture through 100-mesh gauze, and then vacuum drying it at 25° C. to finally obtain a Chinese fir callus extract filtrate.

[0033] As a preferred technical solution of the present invention, the filtrate extracted from Chinese fir callus can effectively promote the proliferation of human cells, especially increase the growth of fibroblasts in the dermis, stimulate collagen synthesis, and promote skin regeneration and repair.

[0034] Beneficial effects of the present invention:

[0035] (1) The present invention uses liposomes prepared from soybean lecithin and cholesterol to encapsulate hesperidin and the reaction product of citral and chitosan oligosaccharide, thereby improving the bioavailability of chitosan oligosaccharide, hesperidin and citral; small molecular weight chitosan oligosaccharide has the functions of inhibiting the growth and reproduction of pathogens, promoting protein synthesis, activating cells, thereby promoting rapid cell growth, and being more easily absorbed by the human body, having a higher bioavailability

[0036] (2) The present invention increases stability and antibacterial properties by reacting citral with chitosan oligosaccharide. Ergothioneine and the composite antioxidant have a synergistic antibacterial mechanism, and the combination can improve the antibacterial activity. The antibacterial properties of the composition are significantly improved by the synergistic effect of hesperidin, chitosan oligosaccharide and citral. DETAILED DESCRIPTION

[0037] In order to further illustrate the technical means and effects adopted by the present invention to achieve the predetermined invention purpose, the specific implementation methods, structures, features and effects of the present invention are described in detail below in combination with the embodiments. Example 1

[0038] A skin repairing composition containing plant callus tissue comprises the following components in parts by weight:

[0039] 7 parts of fir callus extract, 2 parts of ergothioneine, 3 parts of conopeptide, 4 parts of squalane and 3.5 parts of sustained-release material;

[0040] The preparation method of the sustained-release material comprises the following steps:

[0041] S1. Soybean lecithin, cholesterol and anhydrous ethanol are mixed, ultrasonically treated and evaporated to obtain liposomes; wherein the mass ratio of the soybean lecithin to cholesterol is 4.0:1.0; the ultrasonic treatment is performed at a frequency of 100 KHz and for 30 minutes; and the evaporation dispersion is performed at a speed of 50 r / min and a temperature of 30°C.

[0042] S2, after mixing hesperidin, compound antioxidant and liposome, heating until the liposome is hydrated, ultrasonically vibrating, freeze-drying, and obtaining a sustained-release material; wherein the mass ratio of hesperidin, compound antioxidant and liposome is 0.5:1.4:3.2; the conditions of the ultrasonic oscillation are 200r / min speed, 300MHz frequency, and treatment for 15min;

[0043] The preparation method of the composite antioxidant comprises the following steps:

[0044] 1) After chitosan, acetic acid and deionized water are stirred and mixed, hydrogen peroxide is added and stirred evenly for degradation treatment to obtain a chitosan oligosaccharide mixed solution; wherein the mass ratio of the chitosan, acetic acid, deionized water and hydrogen peroxide is 3:4:120:7; the molecular weight of the chitosan is 120 kDa; the degradation treatment conditions are stirring at a speed of 300 r / min, an ultrasonic frequency of 65 KHz, a temperature of 50°C, and degradation for 4 hours; the molecular weight of the chitosan oligosaccharide in the chitosan oligosaccharide mixed solution is 2000;

[0045] 2) In an inert atmosphere, stir and mix citral and ethanol, then add ammonium cerium nitrate solution and chitosan oligosaccharide mixture in sequence, stir and react, add acetone for precipitation, let stand, filter, wash and dry to obtain a composite antioxidant; wherein the amount ratio of citral, ethanol, ammonium cerium nitrate solution and chitosan oligosaccharide mixture is 3.0g:22mL:0.5g:310g; the ethanol is anhydrous ethanol; the inert atmosphere is nitrogen atmosphere; the stirring reaction conditions are 30KHz ultrasonic frequency treatment for 5min, stirring at a speed of 200r / min, at a temperature of 35°C, and reacting for 4h; the standing time is 1h; the washing conditions are washing with anhydrous ethanol until neutral; the drying is freeze drying; the concentration of the ammonium cerium nitrate solution is 0.04g / mL;

[0046] The invention discloses a preparation method of a skin repairing composition containing plant callus tissue, comprising weighing each component by weight and mixing the components to obtain the composition. Example 2

[0047] A skin repairing composition containing plant callus tissue comprises the following components in parts by weight:

[0048] 8 parts of fir callus extract, 2.2 parts of ergothioneine, 3.6 parts of conopeptide, 4.4 parts of squalane and 3.8 parts of sustained-release material;

[0049] The preparation method of the sustained-release material comprises the following steps:

[0050] S1. Soybean lecithin, cholesterol and anhydrous ethanol are mixed, ultrasonically treated and evaporated to obtain liposomes; wherein the mass ratio of the soybean lecithin to cholesterol is 4.06:1.04; the ultrasonic treatment condition is at a frequency of 104KHz and ultrasound for 32 minutes; the evaporation dispersion condition is at a speed of 52r / min and a temperature of 31°C.

[0051] S2, after mixing hesperidin, compound antioxidant and liposome, heating until the liposome is hydrated, ultrasonically vibrating, freeze-drying, and obtaining a sustained-release material; wherein the mass ratio of hesperidin, compound antioxidant and liposome is 0.56:1.5:3.4; the conditions of the ultrasonic oscillation are at a speed of 204r / min, a frequency of 320MHz, and a treatment time of 16min;

[0052] The preparation method of the composite antioxidant comprises the following steps:

[0053] 1) After chitosan, acetic acid and deionized water are stirred and mixed, hydrogen peroxide is added and stirred evenly for degradation treatment to obtain a chitosan oligosaccharide mixed solution; wherein the mass ratio of the chitosan, acetic acid, deionized water and hydrogen peroxide is 4:4.4:120:7.4; the molecular weight of the chitosan is 120 kDa; the conditions of the degradation treatment are stirring at a speed of 320 r / min, an ultrasonic frequency of 68 KHz, a temperature of 54° C., and degradation for 5 hours; the molecular weight of the chitosan oligosaccharide in the chitosan oligosaccharide mixed solution is 2100;

[0054] 2) In an inert atmosphere, citral and ethanol are stirred and mixed, and then ammonium cerium nitrate solution and chitosan oligosaccharide mixture are added in sequence and mixed, and after stirring for reaction, acetone is added for precipitation, and the mixture is allowed to stand, filtered, washed, and dried to obtain a composite antioxidant; wherein the amount ratio of citral, ethanol, ammonium cerium nitrate solution and chitosan oligosaccharide mixture is 3.1g:22.6mL:0.56g:314g; the ethanol is anhydrous ethanol; the inert atmosphere is a nitrogen atmosphere; the stirring reaction conditions are 32KHz ultrasonic frequency treatment for 5.6min, stirring at a speed of 220r / min, and reacting at a temperature of 36°C for 4.4h; the standing time is 1.2h; the washing condition is washing with anhydrous ethanol until neutral; the drying is freeze-drying; the concentration of the ammonium cerium nitrate solution is 0.042g / mL;

[0055] The invention discloses a preparation method of a skin repairing composition containing plant callus tissue, comprising weighing each component by weight and mixing the components to obtain the composition. Example 3

[0056] A skin repairing composition containing plant callus tissue comprises the following components in parts by weight:

[0057] 8.6 parts of fir callus extract, 2.4 parts of ergothioneine, 4.2 parts of conopeptide, 4.8 parts of squalane and 4.1 parts of sustained-release material;

[0058] The preparation method of the sustained-release material comprises the following steps:

[0059] S1. Soybean lecithin, cholesterol and anhydrous ethanol are mixed, ultrasonically treated and evaporated to obtain liposomes; wherein the mass ratio of the soybean lecithin to cholesterol is 4.12:1.08; the ultrasonic treatment condition is at a frequency of 108KHz and ultrasound for 34 minutes; the evaporation dispersion condition is at a speed of 54r / min and a temperature of 32°C.

[0060] S2, after mixing hesperidin, compound antioxidant and liposome, heating until the liposome is hydrated, ultrasonically vibrating, freeze-drying, and obtaining a sustained-release material; wherein the mass ratio of hesperidin, compound antioxidant and liposome is 0.62:1.6:3.5; the conditions of the ultrasonic oscillation are at a speed of 208r / min, a frequency of 340MHz, and a treatment time of 17min;

[0061] The preparation method of the composite antioxidant comprises the following steps:

[0062] 1) After chitosan, acetic acid and deionized water are stirred and mixed, hydrogen peroxide is added and stirred evenly for degradation treatment to obtain a chitosan oligosaccharide mixed solution; wherein the mass ratio of the chitosan, acetic acid, deionized water and hydrogen peroxide is 5:4.8:120:7.8; the molecular weight of the chitosan is 130 kDa; the conditions of the degradation treatment are stirring at a speed of 340 r / min, an ultrasonic frequency of 71 KHz, a temperature of 58° C., and degradation for 5.6 hours; the molecular weight of the chitosan oligosaccharide in the chitosan oligosaccharide mixed solution is 2200;

[0063] 2) In an inert atmosphere, citral and ethanol are stirred and mixed, and then ammonium cerium nitrate solution and chitosan oligosaccharide mixture are added in sequence and mixed, and after stirring for reaction, acetone is added for precipitation, and the mixture is allowed to stand, filtered, washed, and dried to obtain a composite antioxidant; wherein the amount ratio of citral, ethanol, ammonium cerium nitrate solution and chitosan oligosaccharide mixture is 3.2g:23.2mL:0.62g:318g; the ethanol is anhydrous ethanol; the inert atmosphere is a nitrogen atmosphere; the stirring reaction conditions are 34KHz ultrasonic frequency treatment for 6.2min, stirring at a speed of 240r / min, and reacting at a temperature of 37°C for 4.8h; the standing time is 1.4h; the washing condition is washing with anhydrous ethanol until neutral; the drying is freeze-drying; the concentration of the ammonium cerium nitrate solution is 0.044g / mL;

[0064] The invention discloses a preparation method of a skin repairing composition containing plant callus tissue, comprising weighing each component by weight and mixing the components to obtain the composition. Example 4

[0065] A skin repairing composition containing plant callus tissue comprises the following components in parts by weight:

[0066] 9.4 parts of fir callus extract, 2.6 parts of ergothioneine, 4.8 parts of conopeptide, 5.2 parts of squalane and 4.4 parts of sustained-release material;

[0067] The preparation method of the sustained-release material comprises the following steps:

[0068] S1. Soybean lecithin, cholesterol and anhydrous ethanol are mixed, ultrasonically treated and evaporated to obtain liposomes; wherein the mass ratio of the soybean lecithin to cholesterol is 4.18:1.12; the ultrasonic treatment condition is at a frequency of 112KHz and ultrasound for 36 minutes; the evaporation dispersion condition is at a speed of 56r / min and a temperature of 33°C.

[0069] S2, after mixing hesperidin, compound antioxidant and liposome, heating until the liposome is hydrated, ultrasonically vibrating, freeze-drying, and obtaining a sustained-release material; wherein the mass ratio of hesperidin, compound antioxidant and liposome is 0.68:1.8:3.7; the conditions of the ultrasonic oscillation are at a speed of 212 r / min, a frequency of 360 MHz, and a treatment time of 18 min;

[0070] The preparation method of the composite antioxidant comprises the following steps:

[0071] 1) After chitosan, acetic acid and deionized water are stirred and mixed, hydrogen peroxide is added and stirred evenly for degradation treatment to obtain a chitosan oligosaccharide mixed solution; wherein the mass ratio of the chitosan, acetic acid, deionized water and hydrogen peroxide is 7:5.2:120:8.2; the molecular weight of the chitosan is 130 kDa; the degradation treatment conditions are stirring at a speed of 360 r / min, an ultrasonic frequency of 74 KHz, a temperature of 62° C., and degradation for 6.4 hours; the molecular weight of the chitosan oligosaccharide in the chitosan oligosaccharide mixed solution is 2300;

[0072] 2) In an inert atmosphere, citral and ethanol are stirred and mixed, and then ammonium cerium nitrate solution and chitosan oligosaccharide mixture are added in sequence and mixed, and after stirring for reaction, acetone is added for precipitation, and the mixture is allowed to stand, filtered, washed, and dried to obtain a composite antioxidant; wherein the amount ratio of citral, ethanol, ammonium cerium nitrate solution and chitosan oligosaccharide mixture is 3.3g:23.8mL:0.68g:322g; the ethanol is anhydrous ethanol; the inert atmosphere is a nitrogen atmosphere; the stirring reaction conditions are 36KHz ultrasonic frequency treatment for 6.8min, stirring at a speed of 260r / min, at a temperature of 38°C, and reacting for 5.2h; the standing time is 1.6h; the washing condition is washing with anhydrous ethanol until neutral; the drying is freeze-drying; the concentration of the ammonium cerium nitrate solution is 0.046g / mL;

[0073] The invention discloses a preparation method of a skin repairing composition containing plant callus tissue, comprising weighing each component by weight and mixing the components to obtain the composition. Example 5

[0074] A skin repairing composition containing plant callus tissue comprises the following components in parts by weight:

[0075] 10 parts of fir callus extract, 2.8 parts of ergothioneine, 5.4 parts of conopeptide, 5.6 parts of squalane and 4.7 parts of sustained-release material;

[0076] The preparation method of the sustained-release material comprises the following steps:

[0077] S1. Soybean lecithin, cholesterol and anhydrous ethanol are mixed, ultrasonically treated and evaporated to obtain liposomes; wherein the mass ratio of the soybean lecithin to cholesterol is 4.24:1.16; the ultrasonic treatment condition is at a frequency of 116KHz and ultrasound for 38 minutes; the evaporation dispersion condition is at a speed of 58r / min and a temperature of 34°C.

[0078] S2, after mixing hesperidin, compound antioxidant and liposome, heating until the liposome is hydrated, ultrasonically vibrating, freeze-drying, and obtaining a sustained-release material; wherein the mass ratio of hesperidin, compound antioxidant and liposome is 0.74:1.9:3.8; the conditions of the ultrasonic oscillation are at a speed of 216 r / min, a frequency of 380 MHz, and a treatment time of 19 min;

[0079] The preparation method of the composite antioxidant comprises the following steps:

[0080] 1) After chitosan, acetic acid and deionized water are stirred and mixed, hydrogen peroxide is added and stirred evenly for degradation treatment to obtain a chitosan oligosaccharide mixed solution; wherein the mass ratio of the chitosan, acetic acid, deionized water and hydrogen peroxide is 8:5.6:120:8.6; the molecular weight of the chitosan is 140 kDa; the conditions of the degradation treatment are stirring at a speed of 380 r / min, an ultrasonic frequency of 77 KHz, a temperature of 66° C., and degradation for 7 hours; the molecular weight of the chitosan oligosaccharide in the chitosan oligosaccharide mixed solution is 2400;

[0081] 2) In an inert atmosphere, citral and ethanol are stirred and mixed, and then ammonium cerium nitrate solution and chitosan oligosaccharide mixture are added in sequence and mixed, and after stirring for reaction, acetone is added for precipitation, and the mixture is allowed to stand, filtered, washed, and dried to obtain a composite antioxidant; wherein the amount ratio of citral, ethanol, ammonium cerium nitrate solution and chitosan oligosaccharide mixture is 3.4g:24.4mL:0.74g:326g; the ethanol is anhydrous ethanol; the inert atmosphere is a nitrogen atmosphere; the stirring reaction conditions are 38KHz ultrasonic frequency treatment for 7.4min, stirring at a speed of 280r / min, at a temperature of 39°C, and reacting for 5.6h; the standing time is 1.8h; the washing condition is washing with anhydrous ethanol until neutral; the drying is freeze-drying; the concentration of the ammonium cerium nitrate solution is 0.048g / mL;

[0082] The invention discloses a preparation method of a skin repairing composition containing plant callus tissue, comprising weighing each component by weight and mixing the components to obtain the composition. Example 6

[0083] A skin repairing composition containing plant callus tissue comprises the following components in parts by weight:

[0084] 11 parts of fir callus extract, 3 parts of ergothioneine, 6 parts of conopeptide, 6 parts of squalane and 5.0 parts of sustained-release material;

[0085] The preparation method of the sustained-release material comprises the following steps:

[0086] S1. Soybean lecithin, cholesterol and anhydrous ethanol are mixed, ultrasonically treated and evaporated to obtain liposomes; wherein the mass ratio of the soybean lecithin to cholesterol is 4.3:1.2; the ultrasonic treatment condition is 120KHz frequency and 40min; the evaporation dispersion condition is 60r / min rotation speed and 35°C temperature.

[0087] S2, after mixing hesperidin, compound antioxidant and liposome, heating until the liposome is hydrated, ultrasonically vibrating, freeze-drying, and obtaining a sustained-release material; wherein the mass ratio of hesperidin, compound antioxidant and liposome is 0.8:2.0:4.0; the conditions of the ultrasonic oscillation are 220r / min speed, 400MHz frequency, and treatment for 20min;

[0088] The preparation method of the composite antioxidant comprises the following steps:

[0089] 1) After chitosan, acetic acid and deionized water are stirred and mixed, hydrogen peroxide is added and stirred evenly for degradation treatment to obtain a chitosan oligosaccharide mixed solution; wherein the mass ratio of the chitosan, acetic acid, deionized water and hydrogen peroxide is 9:6:120:9; the molecular weight of the chitosan is 150 kDa; the degradation treatment conditions are stirring at a speed of 400 r / min, an ultrasonic frequency of 80 KHz, a temperature of 70°C, and degradation for 8 hours; the molecular weight of the chitosan oligosaccharide in the chitosan oligosaccharide mixed solution is 2500;

[0090] 2) In an inert atmosphere, stir and mix citral and ethanol, then add ammonium cerium nitrate solution and chitosan oligosaccharide mixture in sequence, stir and react, add acetone for precipitation, let stand, filter, wash and dry to obtain a composite antioxidant; wherein the amount ratio of citral, ethanol, ammonium cerium nitrate solution and chitosan oligosaccharide mixture is 3.5g:25mL:0.8g:330g; the ethanol is anhydrous ethanol; the inert atmosphere is nitrogen atmosphere; the stirring reaction conditions are 40KHz ultrasonic frequency treatment for 8min, stirring at a speed of 300r / min, at a temperature of 40°C, and reacting for 6h; the standing time is 2h; the washing condition is washing with anhydrous ethanol until neutral; the drying is freeze drying; the concentration of the ammonium cerium nitrate solution is 0.05g / mL;

[0091] The invention discloses a preparation method of a skin repairing composition containing plant callus tissue, comprising weighing each component by weight and mixing the components to obtain the composition.

[0092] Comparative Example 1

[0093] Compared with Example 3, difference is that comparative example 1 does not use thioneine, and all the other components, preparation process and parameters are consistent.

[0094] Comparative Example 2

[0095] Compared with Example 3, the difference is that Comparative Example 2 does not use hesperidin, and the other components, preparation steps and parameters are the same.

[0096] Comparative Example 3

[0097] Compared with Example 3, the difference is that citral is not used in Comparative Example 3, and the other components, preparation steps and parameters are the same.

[0098] Comparative Example 4

[0099] Compared with Example 3, the difference is that comparative example 4 uses citral instead of the composite antioxidant, and the other components, preparation steps and parameters are the same.

[0100] Comparative Example 5

[0101] Compared with Example 3, the difference is that the molecular weight of the chitosan oligosaccharide in the chitosan oligosaccharide mixed solution of Comparative Example 5 is 5000; the other components, preparation steps and parameters are the same.

[0102] Comparative Example 6

[0103] Compared with Example 3, the difference is that the stirring reaction conditions of step 2) of comparative example 6 are: after 6.2 minutes of ultrasonic treatment at 34KHz frequency, stirring at a speed of 240r / min, at a temperature of 50°C, and reacting for 4.8 hours; the other components, preparation steps and parameters are the same.

[0104] Comparative Example 7

[0105] Compared with Example 3, the difference is that the stirring reaction conditions of step 2) of comparative example 7 are: after 6.2 minutes of ultrasonic treatment at 34KHz frequency, stirring at a speed of 240r / min, at a temperature of 20°C, and reacting for 4.8 hours; the other components, preparation steps and parameters are the same.

[0106] Antibacterial performance test:

[0107] The experiment selected common wound bacteria (Staphylococcus aureus ATCC25923, Escherichia coli ATCC25922, Candida albicans ATCC14053) as the research objects. The activated Staphylococcus aureus, Escherichia coli and Candida albicans were inoculated on ordinary broth medium agar plates and cultured at 37°C for 24 hours. Subsequently, the bacteria were appropriately diluted with nutrient broth, and the compositions of Examples 1-6 and Comparative Examples 1-7 of the present invention were added and incubated for 90 minutes. 50 μL of the sample was taken out for plate coating, and then inverted and cultured at 37°C for 24 hours to observe the growth of the colonies. Sterile water was used as a blank control to calculate the antibacterial rate of each embodiment and comparative example, and the results are shown in Table 1.

[0108] DPPH free radical scavenging test

[0109] In a 10mL colorimetric tube, 4.0mL of DPPH solution and the sample to be tested (Example 1-6, Comparative Example 1-7) were added in sequence, and anhydrous ethanol was added to the scale. After mixing, the absorbance (A) was measured at a wavelength of 517nm using a 1cm colorimetric dish immediately. The absorbance was recorded as Ai. The absorbance was measured after being placed at room temperature and protected from light for 30min, and recorded as Aj. The blank control was only added with DPPH ethanol solution, and its absorbance was recorded as Ac. The DPPH free radical scavenging rate K (%) was calculated according to the following formula; the results are shown in Table 1.

[0110] K=[1-(Ai-Aj) / Ac]*100%;

[0111] Table 1

[0112] Staphylococcus aureus inhibition rate (%) Escherichia coli inhibition rate (%) Candida albicans (%) DPPH free radical scavenging rate (%) Example 1 95.4 96.6 98.7 92.7 Example 2 95.5 97.7 99.5 93.6 Example 3 95.7 97.2 99.6 94.0 Example 4 95.4 96.5 99.4 92.8 Example 5 95.2 96.3 98.7 92.5 Example 6 95.3 96.4 98.8 92.6 Comparative Example 1 82.9 86.3 88.7 83.4 Comparative Example 2 78.0 81.3 83.6 78.6 Comparative Example 3 79.7 82.8 85.2 80.2 Comparative Example 4 76.2 79.4 81.5 76.7 Comparative Example 5 86.3 89.6 91.8 86.5 Comparative Example 6 86.2 87.4 89.5 84.4 Comparative Example 7 85.1 88.7 90.9 85.6

[0113] From the test results in Table 1, it can be seen that compared with Comparative Examples 1-7, the antibacterial properties of the compositions prepared in Examples 1-6 are significantly better than those in Comparative Examples 1-7.

[0114] The above description is only a preferred embodiment of the present invention and does not limit the present invention in any form. Although the present invention has been disclosed as a preferred embodiment as above, it is not used to limit the present invention. Any technical personnel in this field can make some changes or modify the technical contents disclosed above into equivalent embodiments without departing from the scope of the technical solution of the present invention. However, any brief modifications, equivalent changes and modifications made to the above embodiments based on the technical essence of the present invention without departing from the content of the technical solution of the present invention are still within the scope of the technical solution of the present invention.

Claims

1. A skin repair composition containing plant callus, characterized in that: The components include the following by weight: 7-11 parts of fir callus extract, 2-3 parts of ergothioneine, 3-6 parts of conopeptide, 4-6 parts of squalane and 3.5-5.0 parts of sustained-release material; The preparation method of the sustained-release material comprises the following steps: S1. Soybean lecithin, cholesterol and anhydrous ethanol are mixed, subjected to ultrasonic treatment and evaporated dispersion to obtain liposomes; the mass ratio of soybean lecithin to cholesterol is 4.0-4.3:1.0-1.2; the ultrasonic treatment condition is at a frequency of 100-120KHz and an ultrasonic wave for 30-40min; the evaporation dispersion condition is at a speed of 50-60r / min and a temperature of 30-35°C; S2, after mixing hesperidin, compound antioxidant and liposome, heating until the liposome is hydrated, ultrasonic oscillation, freeze-drying, and obtaining a sustained-release material; the mass ratio of hesperidin, compound antioxidant and liposome is 0.5-0.8:1.4-2.0:3.2-4.0; the conditions of the ultrasonic oscillation are 200-220r / min speed, 300-400MHz frequency, and treatment for 15-20min; The preparation method of the composite antioxidant comprises the following steps: 1) After chitosan, acetic acid and deionized water are stirred and mixed, hydrogen peroxide is added and stirred evenly for degradation treatment to obtain a chitosan oligosaccharide mixed solution; the mass ratio of chitosan, acetic acid, deionized water and hydrogen peroxide is 3-9:4-6:120:7-9; the molecular weight of the chitosan is 120-150 kDa; the conditions of the degradation treatment are stirring at a speed of 300-400 r / min, an ultrasonic frequency of 65-80 KHz, a temperature of 50-70° C., and degradation for 4-8 hours; the molecular weight of the chitosan oligosaccharide in the chitosan oligosaccharide mixed solution is 2000-2500; 2) Under an inert atmosphere, stir and mix citral and ethanol, add ammonium cerium nitrate solution and chitosan oligosaccharide mixture in sequence, stir and react, add acetone for precipitation, let stand, filter, wash and dry to obtain a composite antioxidant; the amount ratio of citral, ethanol, ammonium cerium nitrate solution and chitosan oligosaccharide mixture is 3.0-3.5g:22-25mL:0.5-0.8g:310-330g; the ethanol is anhydrous ethanol; the inert atmosphere is nitrogen atmosphere; the stirring reaction conditions are 30-40KHz ultrasonic frequency treatment for 5-8min, stirring at a speed of 200-300r / min, at a temperature of 35-40°C, and reacting for 4-6h; the concentration of the ammonium cerium nitrate solution is 0.04-0.05g / mL; the standing time is 1-2h; the washing condition is washing with anhydrous ethanol until neutral; and the drying is freeze-drying.

2. A method for preparing a skin repair composition containing plant callus as claimed in claim 1, characterized in that: The preparation method comprises the following steps: weighing each component by weight and mixing them to obtain the product.