CN110742853A

CN110742853A - Extracellular matrix freeze-dried powder composition and application thereof

Info

Publication number: CN110742853A
Application number: CN201911132120.6A
Authority: CN
Inventors: 蓝锡伟
Original assignee: Hangzhou Sange Biotechnology Co Ltd
Current assignee: Hangzhou Sange Biotechnology Co Ltd
Priority date: 2019-11-19
Filing date: 2019-11-19
Publication date: 2020-02-04

Abstract

The invention discloses an extracellular matrix freeze-dried powder composition and application thereof. The extracellular matrix freeze-dried powder composition comprises freeze-dried powder and a solvent; the freeze-dried powder comprises the following raw materials: extracellular matrix extract, mannitol, oligopeptide-1, oligopeptide-3, pomegranate bark microcapsule powder and water; the solvent comprises the following raw materials: tremella extract, glycerin and water. When in use, the solvent and the freeze-dried powder are mixed uniformly, are directly dripped on the face after being completely dissolved, and the face skin is gently massaged until the mixture is absorbed; or the solution after complete solution is added into skin care products (water and milk) and is uniformly smeared on the skin, so that the problems of rough skin, fine wrinkles and the like can be effectively improved, the epidermis is repaired, the skin aging is delayed, and the skin is more moist, transparent and smooth.

Description

Extracellular matrix freeze-dried powder composition and application thereof

Technical Field

The invention relates to an extracellular matrix freeze-dried powder composition and application thereof, belonging to the technical field of cosmetics.

Background

The skin needs to be repaired every day, and natural wind, frost, snow, rain, ultraviolet light, serious environmental pollution, food additives, functional components in skin care products, fruit acid hormones, lead and mercury, antibiotics, pigments and the like can damage the self resistance and immunity of the skin, cause damage to the health of the skin to different degrees and cause the problems of premature aging, wrinkles, color spots, acne, dryness, sensitivity, red blood streak and the like of the skin. Therefore, timely repair is needed when skin damage occurs, and the repair is beneficial and harmless for preventing and treating various skin problems.

The true repair product should be safe and effective, be used for a long period of time, and be a product of any brand of care and efficacy, capable of preventing and timely repairing possible skin damage. Therefore, the ecological restoration skin care product without adding any preservative should be selected judiciously, and is a great invention in the field of skin care nowadays.

At present, in the research of cosmetics, freeze-dried powder cosmetics appear, when the freeze-dried powder cosmetics are used, the freeze-dried powder cosmetics need to be matched with water liquid for dissolving freeze-dried powder, and when the existing water liquid for dissolving freeze-dried powder is matched with the freeze-dried powder cosmetics, the skin injury can be repaired. However, the traditional freeze-dried powder has poor repairing effect on skin, and the duration time and the duration effect of the repairing effect are not ideal.

The pomegranate rind extract powder microcapsule is added on the basis of the extracellular matrix extract, so that the stability of the effective components is obviously improved, the effect of the effective components is greatly exerted, and the application of the extracellular matrix freeze-dried powder in whitening and skin care is improved.

Disclosure of Invention

The invention aims to provide an extracellular matrix freeze-dried powder composition which can be applied to the field of whitening and skin care, effectively improve the problems of rough skin, fine wrinkles and the like, repair epidermis, delay skin aging and enable the skin to be more moist, transparent and smooth.

The specific technical method comprises the following steps:

an extracellular matrix lyophilized powder composition comprises lyophilized powder and solvent; the mass volume ratio of the freeze-dried powder to the solvent is (15-25) to 1 g/L; the freeze-dried powder comprises the following raw materials: the extracellular matrix extract, mannitol, oligopeptide-1, oligopeptide-3 and water are prepared from the following raw materials: tremella extract, glycerin and water. When in use, the solvent and the freeze-dried powder are uniformly mixed and completely dissolved to obtain the extracellular matrix freeze-dried powder composition, and then the composition is directly dripped on the face, and the face skin is lightly massaged until the composition is absorbed; or adding the solution into skin care product (water, milk), and uniformly spreading on skin.

Oligopeptide-1 (Oligopeptide-1) is a polymer of glycine, histidine and lysine, and is capable of firming loose skin, increasing elasticity, reducing fine lines and wrinkles, reducing photodamage and pigmentation, and increasing keratinocyte proliferation.

Oligopeptide-3 (Oligopeptide-3), is tridecapeptide composed of alanine, leucine-lysine and phenylalanine.

The other technical scheme is that the extracellular matrix freeze-dried powder composition comprises freeze-dried powder and a solvent; the mass volume ratio of the freeze-dried powder to the solvent is (15-25) to 1 g/L; the freeze-dried powder comprises the following raw materials in percentage by mass: 1-3% of extracellular matrix extract, 4-8% of mannitol, 10.3-1.0% of oligopeptide, 30.3-1.0% of oligopeptide, 0.1-0.5% of pomegranate bark microcapsule powder and the balance of water; the solvent consists of the following raw materials in percentage by mass: 3-10% of tremella extract, 0.5-3% of glycerol and the balance of water.

The preparation method of the extracellular matrix extract comprises the following steps:

(1) wrapping the articular cartilage of the cattle with a polyethylene film, and freezing the articular cartilage of the cattle at the temperature of between 20 ℃ below zero and 10 ℃ below zero for more than 15 hours;

(2) directly grinding the frozen bovine articular cartilage, and mixing the ground material with the liquid material according to the ratio of 1: (0.2-0.5) g/L of the solution is added into the solution A and stirred for 0.5-1.5 h at the temperature of 35-42 ℃; centrifuging for 3-7 min at 1-6 ℃ and 6000-8000 rpm, and taking a precipitate; according to the material-liquid ratio of 1: (0.2-0.5) g/L of the precipitate is added into the solution B and stirred for 20-30 h at 4 ℃; centrifuging for 3-7 min at 1-6 ℃ and 6000-8000 rpm, and taking a precipitate; according to the material-liquid ratio of 1: (0.2-0.5) g/L of the precipitate is added into the solution C and stirred for 10-14 h at the temperature of 35-42 ℃; centrifuging at 1-6 ℃ and 6000-8000 rpm for 3-7 min, taking the precipitate, washing with water for 1-3 times, and carrying out vacuum freeze drying to obtain the extracellular matrix extract.

Solution A: 1 wt% trypsin, 1 wt% ethylenediaminetetraacetic acid, and the balance of 0.01mol/L PBS buffer solution with pH of 7.5;

solution B: 0.1mmol/L phenylmethylsulfonyl fluoride, 1 wt.% ethylenediaminetetraacetic acid, 1 wt.% Triton X-100, and the balance 0.01mol/L Tris-hydrochloric acid buffer solution having a pH of 7.5;

solution C: 50U/mL DNase 1(DNase L), 1U/mL RNase A (RNase A), and the balance 0.01M PBS buffer at pH 7.5.

Preferably, the preparation method of the pomegranate bark microcapsule powder comprises the following steps:

(1) wrapping fresh pomegranate rind with polyethylene film, and freezing at-20-10 deg.c for over 15 hr;

(2) directly grinding the frozen pomegranate rind, adding the ground pomegranate rind into an ethanol hydrochloric acid solution according to the material-liquid ratio of 1 (0.2-0.5) g/L, and performing ultrasonic treatment at 5-30 ℃ for 5-20 min; stirring for 20-30 h in a water bath at 5-30 ℃, filtering, rotatably evaporating the filtrate at 40-50 ℃ to the original volume of 1/8-1/5, carrying out vacuum freeze drying, and grinding into powder to obtain an extract; the hydrochloric acid ethanol solution is formed by mixing 96 wt% ethanol water solution and 1.5mol/L hydrochloric acid according to the volume ratio of 85 (10-20);

(3) uniformly mixing 5-15 parts by weight of maltodextrin, 0.1-0.3 part by weight of calcium alginate, 0.1-0.3 part by weight of Arabic gum powder and 75-95 parts by weight of water, standing at 1-8 ℃ for 20-30 hours to obtain a mixed solution, adding the extract into the mixed solution according to a mass ratio of 1 (3-5), stirring at 5-30 ℃ and 100-300 rpm for 20-40 min, and then performing spray drying to obtain pomegranate bark microcapsule powder; the air inlet temperature of the spray drying is 150-210 ℃, and the air outlet temperature is 70-110 ℃.

Preferably, the preparation method of the tremella extract comprises the following steps:

s1, soaking in water and pulping: adding water with the weight 20-50 times of the dry weight of the dried tremella into the dried tremella, soaking for 5-10 hours, and pulping;

s2 enzyme treatment: adding 0.1-0.5 wt% of trypsin and 0.1-0.5 wt% of amylase into the slurry, and carrying out enzymolysis for 8-10 hours at 50-60 ℃;

s3 heat leaching: then stirring and mixing for 2-4 h at 65-85 ℃ and 100-300 rpm;

s4 filtering, concentrating, and vacuum freeze drying to obtain Tremella extract.

The invention also provides an application of the extracellular matrix freeze-dried powder composition in preparing whitening skin care products, which comprises the following steps: adding 0.1-3.0 wt% of the extracellular matrix freeze-dried powder composition into a whitening skin care product.

Extracellular matrix: animal articular cartilage is composed of a small amount of chondrocytes and a large amount of extracellular matrix, and the extracellular matrix (ACECM) of the articular cartilage mainly contains bioactive components such as collagen and proteoglycan; has good biocompatibility and bioactivity.

The functional polysaccharide in the tremella is α -mannan which has unique smooth skin feeling and high-efficiency moisture and skin care function, so that the functional polysaccharide is known as 'plant hyaluronic acid', α -mannan is rich in polar groups such as hydroxyl, carboxyl and the like in molecules, can combine a large amount of water, is interwoven into a net shape among molecules, has extremely strong water locking and moisture retention performances, exerts high-efficiency moisture and skin care functions, contains rich protein, vitamins and the like, has natural colloid, has the functions of moisturizing skin, resisting aging, removing wrinkles and tightening the skin after long-term use, and has the effects of removing chloasma and freckles on the face.

Pomegranate, sweet in nature and taste, sour and astringent, warm in nature, has the effects of killing parasites, astringing intestines, stopping dysentery and the like. The pomegranate rind contains various chemical components, mainly contains high molecular weight phenols, ellagitannin, proanthocyanidin, complex polysaccharide flavone and the like, and antibacterial tests prove that alcohol extract and peel decoction of the pomegranate have broad-spectrum antibacterial action, and have obvious inhibition effects on staphylococcus aureus, hemolytic streptococcus, vibrio cholerae, dysentery bacillus and the like, and the pomegranate rind water infusion also has inhibition effects on various dermatophytes in test tubes to different degrees. However, the introduction of phenols, anthocyanins, into the cosmetic field presents certain difficulties and technical obstacles, considering their low stability during processing and storage. The invention adopts microencapsulation technology, and biologically active compounds are encapsulated by biological polymers, so that the biologically active compounds are prevented from being influenced by oxygen, water, light or other environmental factors, the stability of the biologically active compounds is improved, and the biologically active compounds are applied to the field of cosmetics.

The invention has the beneficial effects that: the preparation method is characterized in that a microcapsule powdered pomegranate bark extract is added on the basis of the extracellular matrix extract, and the pomegranate bark extract is encapsulated by a biopolymer by adopting a microencapsulation technology, so that the stability of the pomegranate bark extract can be effectively improved, and the oxidation resistance of the extracellular matrix freeze-dried powder is remarkably improved; meanwhile, the skin is repaired, and ecological skin care is realized.

Detailed Description

The preparation method of the extracellular matrix freeze-dried powder in the examples and the comparative examples is as follows:

(1) wrapping the bovine articular cartilage with a polyethylene film, and freezing at-18 ℃ for 24 hours;

(2) directly grinding the frozen bovine articular cartilage, and mixing the ground material with the liquid material according to the ratio of 1: adding 0.4g/L of the solution A into the solution A, and stirring for 1.0h at 37 ℃; centrifuging at 4 deg.C and 7000rpm for 5min, and collecting precipitate; according to the material-liquid ratio of 1: 0.4g/L of the precipitate is added into the solution B and stirred for 24 hours at 4 ℃; centrifuging at 4 deg.C and 7000rpm for 5min, and collecting precipitate; according to the material-liquid ratio of 1: 0.4g/L of the precipitate is added into the solution C and stirred for 12 hours at 37 ℃; centrifuging at 4 deg.C and 7000rpm for 5min, collecting precipitate, washing with water for 3 times, and vacuum freeze drying to obtain extracellular matrix extract.

The dissolving solution A is 1 wt% of trypsin, 1 wt% of ethylenediamine tetraacetic acid and the balance of 0.01mol/L PBS buffer solution with pH of 7.5;

the dissolving solution B is 0.1mmol/L phenylmethylsulfonyl fluoride, 1 wt% ethylene diamine tetraacetic acid, 1 wt% Triton X-100 and the balance of 0.01mol/L Tris-hydrochloric acid buffer solution with pH value of 7.5;

the lysis solution C was 50U/mL of DNase 1(DNase l), 1U/mL of rnase a (rnase a), and the balance was 0.01MpH ═ 7.5 in PBS buffer.

Comparative example 1

An extracellular matrix lyophilized powder composition comprises lyophilized powder and solvent, wherein the mass volume ratio of lyophilized powder to solvent is 20:1g/L, when in use, the solvent and lyophilized powder are uniformly mixed, and are completely dissolved to obtain extracellular matrix lyophilized powder composition, and then the composition is directly dripped on face, and the face skin is gently massaged until absorption.

The freeze-dried powder comprises the following raw materials in percentage by mass: 2% of extracellular matrix extract, 5% of mannitol, 10.5% of oligopeptide, 30.8% of oligopeptide, 0.3% of pomegranate bark extract and the balance of water.

The solvent consists of the following raw materials in percentage by mass: 5% of tremella extract, 1.5% of glycerol and the balance of water.

The preparation method of the pomegranate bark extract comprises the following steps:

(1) wrapping fresh pomegranate rind with 140 μm polyethylene film, and freezing at-18 deg.C for 24 hr;

(2) grinding the frozen pomegranate rind to 2mm by using a grinder, adding the ground pomegranate rind into a hydrochloric acid ethanol solution according to the material-liquid ratio of 1:0.25g/L, and performing ultrasonic treatment at 25 ℃ for 10 min; stirring in water bath at 25 deg.C for 24 hr, filtering, rotary evaporating the filtrate at 45 deg.C to original volume of 1/6, vacuum freeze drying, and grinding into powder to obtain extract; the hydrochloric acid ethanol solution is formed by mixing 96 wt% ethanol water solution and 1.5mol/L hydrochloric acid according to the volume ratio of 85: 15;

(3) uniformly mixing 12 parts by weight of maltodextrin, 0.4 part by weight of calcium alginate and 88 parts by weight of water, standing at 4 ℃ for 24 hours to obtain a mixed solution, adding the extract into the mixed solution according to the mass ratio of 1:5, stirring for 30min at 25 ℃ and 100rpm, and then carrying out spray drying to obtain pomegranate peel microcapsule powder; the air inlet temperature of the spray drying is 180 ℃, and the air outlet temperature is 80 ℃.

The preparation method of the tremella extract comprises the following steps:

s1, soaking in water and pulping: adding water with the weight 25 times of the dry weight of the dried white fungus into the dried white fungus, soaking for 8 hours, and pulping by using a grinder;

s2 enzyme treatment: adding 0.1-0.5 wt% of trypsin and 0.1-0.5 wt% of amylase into the slurry for enzymolysis for 9 hours at 55 ℃;

s3 heat leaching: then stirring and mixing the mixture for 3 hours at 80 ℃ and 300 rpm;

s4 filtration, concentration, vacuum freeze drying: filtering with 500 mesh gauze, rotary evaporating the filtrate at 45 deg.C to 1/8, vacuum freeze drying, and grinding into powder to obtain Tremella extract.

Example 1

The freeze-dried powder comprises the following raw materials in percentage by mass: 2% of extracellular matrix extract, 5% of mannitol, 10.5% of oligopeptide, 30.8% of oligopeptide, 0.3% of pomegranate bark microcapsule powder and the balance of water.

The preparation method of the pomegranate bark microcapsule powder comprises the following steps:

The preparation method of the tremella extract comprises the following steps:

Example 2

(3) uniformly mixing 12 parts by weight of maltodextrin, 0.4 part by weight of Arabic gum powder and 88 parts by weight of water, standing at 4 ℃ for 24 hours to obtain a mixed solution, adding the extract into the mixed solution according to the mass ratio of 1:5, stirring for 30min at 25 ℃ and 100rpm, and then carrying out spray drying to obtain pomegranate peel microcapsule powder; the air inlet temperature of the spray drying is 180 ℃, and the air outlet temperature is 80 ℃.

The preparation method of the tremella extract comprises the following steps:

Example 3

(3) uniformly mixing 12 parts by weight of maltodextrin, 0.2 part by weight of calcium alginate, 0.2 part by weight of Arabic gum powder and 88 parts by weight of water, standing at 4 ℃ for 24 hours to obtain a mixed solution, adding the extract into the mixed solution according to the mass ratio of 1:5, stirring at 25 ℃ and 100rpm for 30min, and then carrying out spray drying to obtain pomegranate bark microcapsule powder; the air inlet temperature of the spray drying is 180 ℃, and the air outlet temperature is 80 ℃.

The preparation method of the tremella extract comprises the following steps:

Test example 1

The extracellular matrix lyophilized powder of examples 1 to 3 and comparative example 1 was tested for its ability to scavenge DPPH free radicals: preparing a sample solution with the concentration of 0.5mg/mL by using absolute ethyl alcohol as a solvent; to a 1cm quartz cuvette, 2.0mL of the sample solution and 2.0mL of a 1.0X 10 molar concentration⁴Uniformly mixing a mol/L DPPH solution, and then placing the mixture in a dark place for 30 minutes; to be provided withTaking an ethanol water solution with the volume fraction of 95% as a reference solution, and measuring a light absorption value A at 517 nm; the same method is used for measuring the light absorption value A of a mixed solution of 2.0mL of sample solution and 2.0mL of ethanol water solution with the volume fraction of 95 percent at 517nm_o(ii) a Further, 2.0mL of the solution was measured to have a molar concentration of 1.0X 10⁴The light absorption value A of a mixed solution of a DPPH solution of mol/L and an ethanol aqueous solution of 2.0mL with the volume fraction of 95 percent at 517nm₁The inhibition rate was calculated according to the following formula:

inhibition ratio (%) {1- (A-A)_o)/A₁}×100。

The specific test results are shown in table 1.

Table 1: DPPH cleaning results Table

	Inhibition ratio (%)
		Comparative example 1	42.31
Example 1	48.32
		Example 2	49.57
Example 3	59.72

Test example 2

The extracellular matrix lyophilized powders of examples 1 to 3 and comparative example 1 were tested for their moisturizing effect.

Selecting 80 persons 20-50 years old, half of men and women, keeping healthy, having regular life and no skin diseases, randomly dividing into 4 groups, wherein each group comprises 20 persons, each group respectively uses the extracellular matrix freeze-dried powder obtained in the embodiment to be uniformly mixed, the extracellular matrix freeze-dried powder is directly dripped on the face after being completely dissolved, the skin of the face is gently massaged until being absorbed, bathing and washing the face after 1h, then measuring the water content of the left face in an environment with the test environment temperature of 25 ℃ and the relative humidity of 60%, and simultaneously keeping the tested person calm. The left face was marked with 2 square test areas of 2cm × 2cm, and the skin moisture content 2 hours after washing the face with a bath was measured with a skin moisture meter, and the results were recorded. The specific results are shown in Table 2.

Table 2: moisturizing effect test result table

Test example 3

Skin elasticity test and evaluation:

and (3) judging by using a skin tester, measuring the skin elasticity value by using the skin tester before using the extracellular matrix freeze-dried powder, and measuring the skin elasticity value by using the skin tester after using the extracellular matrix freeze-dried powder for 15 days.

Skin elasticity increase rate (skin elasticity increase rate after test-skin elasticity before test)/skin elasticity before test)

Table 3: skin elasticity test result table

	The rate of increase in skin elasticity%
		ControlExample 1	17.5
Example 1	25.4
		Example 2	28.6
Example 3	35.2

The present invention has been described in detail, and the principle and embodiments of the present invention are explained herein by using specific examples, which are only used to help understand the method and the core idea of the present invention; meanwhile, for a person skilled in the art, according to the idea of the present invention, there may be variations in the specific embodiments and the application scope, and in summary, the content of the present specification should not be construed as a limitation to the present invention.

Claims

1. An extracellular matrix freeze-dried powder composition is characterized in that: comprises freeze-dried powder and a solvent; the mass-volume ratio of the freeze-dried powder to the solvent is (15-25) to 1 g/L.

2. The extracellular matrix lyophilized powder composition of claim 1, wherein the lyophilized powder comprises the following raw materials: extracellular matrix extract, mannitol, oligopeptide-1, oligopeptide-3 and water.

3. The extracellular matrix lyophilized powder composition of claim 1, wherein the vehicle comprises the following raw materials: tremella extract, glycerin and water.

4. The extracellular matrix lyophilized powder composition of claim 2, wherein the lyophilized powder is composed of the following raw materials by mass: 1-3% of extracellular matrix extract, 4-8% of mannitol, 10.3-1.0% of oligopeptide, 30.3-1.0% of oligopeptide, 0.1-0.5% of pomegranate bark microcapsule powder and the balance of water.

5. The extracellular matrix lyophilized powder composition of claim 3, wherein the solvent comprises the following raw materials by mass: 3-10% of tremella extract, 0.5-3% of glycerol and the balance of water.

6. The extracellular matrix lyophilized powder composition of claim 4, wherein the pomegranate rind microcapsule powder is prepared by the following method:

(2) directly grinding the frozen pomegranate rind, adding the ground pomegranate rind into an ethanol hydrochloric acid solution according to the material-liquid ratio of 1 (0.2-0.5) g/L, and performing ultrasonic treatment at 5-30 ℃ for 5-20 min; stirring for 20-30 h in a water bath at 5-30 ℃, filtering, rotatably evaporating the filtrate at 40-50 ℃ to 1/8-1/5 of the original volume, freeze-drying in vacuum, and grinding into powder to obtain an extract; the hydrochloric acid ethanol solution is formed by mixing 96 wt% ethanol water solution and 1.5mol/L hydrochloric acid according to the volume ratio of 85 (10-20);

(3) uniformly mixing 5-15 parts by weight of maltodextrin, 0.1-0.3 part by weight of calcium alginate, 0.1-0.3 part by weight of Arabic gum powder and 75-95 parts by weight of water, standing at 1-8 ℃ for 20-30 hours to obtain a mixed solution, adding the extract into the mixed solution according to the mass ratio of 1 (3-5), stirring at 5-30 ℃ and 100-300 rpm for 20-40 min, and then performing spray drying to obtain the pomegranate rind microcapsule powder.

7. An extracellular matrix lyophilized powder composition according to claim 6, wherein: the air inlet temperature of the spray drying is 150-210 ℃, and the air outlet temperature is 70-110 ℃.

8. The extracellular matrix lyophilized powder composition according to claim 3 or 5, wherein the preparation method of the tremella extract comprises:

9. The application of the extracellular matrix freeze-dried powder composition in preparing whitening skin care products is characterized in that: adding 0.1-3.0 wt% of extracellular matrix lyophilized powder composition according to any one of claims 1-8 to a whitening skin care product.