CN114569536B - Emulsion with effect of improving microecology and physiological state of aging skin - Google Patents
Emulsion with effect of improving microecology and physiological state of aging skin Download PDFInfo
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/99—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from microorganisms other than algae or fungi, e.g. protozoa or bacteria
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/33—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing oxygen
- A61K8/34—Alcohols
- A61K8/345—Alcohols containing more than one hydroxy group
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/33—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing oxygen
- A61K8/37—Esters of carboxylic acids
- A61K8/375—Esters of carboxylic acids the alcohol moiety containing more than one hydroxy group
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/60—Sugars; Derivatives thereof
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/92—Oils, fats or waxes; Derivatives thereof, e.g. hydrogenation products thereof
- A61K8/922—Oils, fats or waxes; Derivatives thereof, e.g. hydrogenation products thereof of vegetable origin
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
- A61K8/9706—Algae
- A61K8/9711—Phaeophycota or Phaeophyta [brown algae], e.g. Fucus
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
- A61K8/9728—Fungi, e.g. yeasts
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
- A61K8/9783—Angiosperms [Magnoliophyta]
- A61K8/9789—Magnoliopsida [dicotyledons]
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q17/00—Barrier preparations; Preparations brought into direct contact with the skin for affording protection against external influences, e.g. sunlight, X-rays or other harmful rays, corrosive materials, bacteria or insect stings
- A61Q17/005—Antimicrobial preparations
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
- A61Q19/08—Anti-ageing preparations
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/40—Chemical, physico-chemical or functional or structural properties of particular ingredients
- A61K2800/59—Mixtures
- A61K2800/592—Mixtures of compounds complementing their respective functions
- A61K2800/5922—At least two compounds being classified in the same subclass of A61K8/18
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
- A61K2800/85—Products or compounds obtained by fermentation, e.g. yoghurt, beer, wine
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Abstract
The invention discloses an emulsion with an effect of improving the microecology and the physiological state of the skin of an aging population, which takes a brown algae microzyme fermentation lysate, lactobacillus/pear juice fermentation filtrate and short-chain fatty acid salt as main active ingredients, and can obviously improve the microecology of the skin of the aging population and has an effect of improving the physiological state of the skin of the aging population through population test verification. Meanwhile, the emulsion can strengthen the barrier of the aged skin, promote the synthesis of elastin, enable the product to have the efficacy of anti-wrinkle repair, and can be added into an anti-aging skin care product which is micro-ecologically friendly to the skin.
Description
Technical Field
The invention relates to an emulsion with an effect of improving the microecology and the physiological state of the skin of aged skin, belonging to the technical field of skin care products.
Background
With the increasing aging of the global population, anti-aging is a persistent topic for humans. Research into skin aging and skin rejuvenation is also becoming a research hotspot. The skin is the largest organ for the human body to directly contact with the external environment, and is the first defense line for protecting the body from nutrient loss and defending invasion of external harmful factors. The barrier function of skin is divided into physical, chemical and biological barriers. The biological barrier is used for preventing the colonization of pathogenic microorganism flora and temporary flora and maintaining the steady state of resident flora. The skin microbial flora has important significance for body health, in particular skin health. The skin microbial flora not only helps the body clean, metabolize skin secretion, excretion and metabolism substances, and is converted into substances with nutritive value for the skin to be absorbed and utilized again by the skin, but also can regulate the congenital and adaptive immune systems of the body. With age, i.e. the development of the natural aging process, the skin microbial flora changes significantly.
Skin microorganisms are mainly present on the skin surface and in skin appendages, such as hair follicles, sweat glands, sebaceous glands, etc., with the most representative microorganisms including propionibacteria, corynebacteria, staphylococcus, and malassezia. A plurality of researches show that with the increase of age, the proportion of the firmicutes, actinomycetes, proteus and bacteroides of different age groups is different at the door level; in particular, the proportion of the Proteus in the senior group is reduced from 35% to 21.9% relative to that in the adult group. In the study published in 2017 aperture, the flora of the young and the senior groups were compared, and analysis based on the number of OTUs observed showed that the senior group showed significantly higher species abundance than the young group at all test sites, and that the senior group showed a higher tendency for alpha diversity than the young group in all skin microbiome. At these sites, corynebacteria show higher abundance in the senior group of cheek and forehead microorganisms; propionibacterium shows higher abundance in the young group of cheeks and forehead; staphylococci show higher abundance in the forearm of the young group; streptococci and prasuvorexant show higher abundance in the young group. In another study, age groups of 20-30s, 50-60s had significant differences in gate level, with bacteroides and firmicutes having higher abundance in 20-30s and proteus and actinomycetes having higher abundance in 50-60 s. More and more researches show that the skin microecology is changed continuously with the increase of the age, and the skin microecology is interacted with the skin state. In the latest study, the abundance of 20-29s,40-53s facial skin microbiome flora in korean healthy females was compared, and the difference in relative abundance between the two age groups was the streptococcus, which was in positive correlation with skin elasticity in combination with the difference in skin elasticity index between the two age groups.
The micro-ecology of aged skin is greatly changed compared with young skin, and the physiological state of the skin is affected, including parameters such as elasticity, compactness and sensitivity of the skin. CN20210104295.7 discloses a composition beneficial to skin micro-ecology after staying up, a preparation method and application, which illustrate the change of skin micro-ecology after staying up, and from the balance of flora for improving skin micro-ecology after staying up, an innovative composition beneficial to skin micro-ecology after staying up is obtained. At present, a method for enabling the skin microecology of the aged skin to trend towards the microecology of young skin is not reported, and corresponding raw materials or combined components for improving the microbial flora of the aged skin and verification after the microecology of the aged skin is improved are not studied.
Disclosure of Invention
The invention aims to provide an emulsion with an effect of improving the microecology and the physiological state of the skin. The emulsion contains brown algae yeast fermentation lysate, lactobacillus/pear juice fermentation filtrate, and short chain fatty acid salt as main effective components, and can improve microecology of aging skin and physiological state of skin. Meanwhile, the emulsion can strengthen the barrier of the aged skin, promote the synthesis of elastin, and enable the product to have the efficacy of anti-wrinkle repair.
The technical scheme of the invention is as follows: an emulsion with effect of improving skin microecology and skin physiological state is characterized by comprising at least three effective components of brown algae microzyme fermented lysate, lactobacillus/pear juice fermented filtrate and short-chain fatty acid salt.
The dosage of the three active ingredients in the emulsion is as follows: each 20g of the emulsion contains 0.1-0.3g of brown algae yeast fermentation lysate (calculated by dry matter, the same applies below), 0.5-8.0g of lactobacillus/pear juice fermentation filtrate and 0.05-0.1g of short chain fatty acid salt. Preferably, it is: each 20g of emulsion contains 0.2g of brown algae microzyme fermented lysate, 3.0g of lactobacillus/pear juice fermented filtrate and 0.06g of short-chain fatty acid salt.
The preparation method of the brown algae microzyme fermentation lysate comprises the following steps: inoculating the brown algae aqueous solution into saccharomycetes for fermentation, breaking walls of the fermentation liquor by a high-pressure homogenizer, crushing thalli, and filtering to obtain the brown algae aqueous solution, wherein the mass ratio of brown algae (wet) to water is 1:2-4, preferably 1:3; the yeast is baker's yeast or beer yeast, preferably baker's yeast; the fermentation temperature is 30-38deg.C, preferably 36 deg.C; the fermentation time is 48-96h, preferably 72h; the fermentation liquor has a filtration pore size of 200-400 mesh, preferably 400 mesh.
The preparation method of the lactobacillus/pear juice fermentation filtrate comprises the following steps: fermenting and culturing lactobacillus in pear juice culture medium (MRS culture medium containing 20+ -5% freshly squeezed pear juice), centrifuging, and collecting supernatant which is lactobacillus/pear juice fermentation filtrate containing lactic acid and amino acid as main components, wherein the lactic acid content is 2.0-6.0wt%. The fermentation temperature is 30-38deg.C, preferably 37deg.C; the fermentation time is 24-72h, preferably 48h.
Furthermore, the invention can be added with sucrose ester emulsifier, natural oil, caprylic/capric triglyceride, polyalcohol and water to prepare the optimal emulsion formula, and the components and the dosage are as follows: each 20g of emulsion contains 0.1-0.3g of brown algae microzyme fermentation lysate, 0.5-8.0g of lactobacillus/pear juice fermentation filtrate, 0.05-0.1g of short-chain fatty acid salt, 0.2-2.0g of sucrose ester emulsifying agent, 0.5-10.0g of natural grease, 1.0-4.0g of caprylic/capric triglyceride, 0.5-10.0g of polyalcohol and the balance of water.
Further preferably, the components and the amounts thereof are as follows: every 20g of the emulsion contains 0.2g of brown algae microzyme fermented lysate, 3.0g of lactobacillus/pear juice fermented filtrate, 0.06g of short-chain fatty acid salt, 1.0g of sucrose ester emulsifying agent, 4.0g of natural grease, 2.5g of caprylic/capric triglyceride, 5.5g of polyalcohol and the balance of water.
The short-chain fatty acid salt is one or two of lactate, butyrate, propionate and isobutyrate, preferably calcium propionate.
The sucrose lipid emulsifying agent is one or two of sucrose laurate, sucrose palmitate and sucrose stearate, preferably sucrose laurate.
The natural oil is one or two of Beacon flower seed oil, mala oil, camellia seed oil, coconut oil and bitter almond oil.
The polyalcohol is one or two of octanediol, pentanediol, butanediol, glycerol, dipropylene glycol and hexanediol.
The preparation method of the emulsion is characterized by comprising the following steps:
s1: mixing sucrose ester emulsifier with polyalcohol, heating for dissolving, adding water, and cooling to form liquid I;
s2: mixing natural oil and caprylic/capric triglyceride to form liquid II;
s3: adding the brown algae microzyme fermented lysate, short-chain fatty acid salt and lactobacillus/pear juice fermentation liquor into the mixed liquor of the first liquid, and uniformly stirring and mixing to form a third liquid;
s4: slowly adding the second liquid into the third liquid, and homogenizing at high speed to form uniform emulsion. The high-speed homogenizing speed is 8000-10000r/min, and the homogenizing time is 5-10min.
The emulsion has effect in improving skin microecology and skin physiological state, and can be added into skin care product for resisting aging.
The brown algae yeast fermentation lysate is a product of brown algae through yeast fermentation, more brown algae oligosaccharides can be obtained, the fermented yeast lysate contains substances such as yeast glucan, nucleotide and the like, and the active ingredients can improve the microecology of aged skin and have better skin repairing effect.
The lactobacillus/pear juice fermentation filtrate is obtained by fermenting lactobacillus in pear juice, and utilizes fructose in pear juice to generate certain organic acid such as lactic acid and free amino acid, and the growth of propionibacterium can be well influenced by combining with the added short-chain fatty acid salt. Meanwhile, lactobacillus can decompose the components in pear juice to generate components beneficial to skin.
The emulsion disclosed by the invention takes the brown algae yeast fermentation lysate, the lactobacillus/pear juice fermentation filtrate and the short-chain fatty acid salt as main effective components, and the brown algae oligosaccharides and the yeast glucan contained in the brown algae yeast fermentation lysate, the lactic acid in the lactobacillus/pear juice fermentation filtrate and the short-chain fatty acid salt are used as prebiotics and probiotics of skin flora to act on skin micro-ecology in a synergistic way, so that the growth of the skin flora is influenced, and the trend of obviously adjusting the transformation from the aging skin flora structure to the young skin flora structure is achieved; the metabolic products such as nucleotide in the brown algae microzyme fermentation lysate and various amino acids generated by lactobacillus/pear juice fermentation filtrate can be quickly permeated and absorbed due to small molecular weight, and can be jointly acted on the isolated skin, so that the thickness of the epidermis layer is obviously increased, the expression of the dermis layer elastin is promoted, and the physiological state of the aged skin is improved. Taken together, the invention mainly utilizes the 3 components to improve the microecology of the aged skin and the physiological state of the skin. The sucrose ester emulsifier has sucrose end, can promote the growth of skin probiotics, namely staphylococcus epidermidis, as an influencing component of flora, and has positive regulation effect on skin flora. Sucrose ester emulsifier, natural oil, caprylic/capric triglyceride and polyalcohol can also play roles of moistening, moisturizing, emulsifying, stabilizing, homogenizing and the like, so that the optimal emulsion formula is prepared, and the effective components play the optimal functions.
The invention has the beneficial effects that:
1. the emulsion truly has the effect of improving the skin microecology and the skin physiological state of the aged people, and the crowd test verifies that the emulsion can remarkably improve the skin microecology of the aged people and has the effect of improving the skin physiological state of the aged people.
2. The composition can remarkably increase the thickness of epidermis and promote the expression of dermis elastin.
3. The main efficacy components in the composition are natural sources, are easy to obtain, safe and mild, and are convenient to use.
Drawings
Fig. 1 is a graph showing facial physiological index comparisons of 262 healthy females with different aging degrees, and is annotated: in this study, x: p is more than 0.01 and less than or equal to 0.05; * *: p is more than 0.001 and less than or equal to 0.01; * **: p is less than or equal to 0.001;
FIG. 2 is a bar graph (genus level) of bacterial species composition Chao index (A) and Shannon index (B);
FIG. 3 is a graph of the composition (genus level) and composition differences of different groups of bacteria species;
FIG. 4 is a graph showing the effect of different samples on EX vivo skin tissue morphology;
FIG. 5 is a graph showing the effect of different samples on EX vivo skin tissue elastin expression;
FIG. 6 shows the composition (genus level) of skin bacteria species of a tester after 7 days of use;
FIG. 7 is a comparison (genus level) of the relative abundance of skin bacterial species before and after use of the test product;
fig. 8 is a graph showing changes in skin wrinkles SEw and dermis layer echo intensity before and after use of the test product: the significance labeling method comprises the following steps: vs. D0; "n.s." means no statistical difference, p >0.05; ". Times." indicates significant differences, p <0.05 is 0.0001;
FIG. 9 is a graph of the variation of sipe length and sipe count before and after use of a test product; the significance labeling method comprises the following steps: vs. D0; "n.s." means no statistical difference, p >0.05; ". Times." indicates significant differences, p <0.05 is 0.0001;
FIG. 10 is a graph showing the change in color a of the subcutaneous red area before and after use of the test product; the significance labeling method comprises the following steps: vs. D0; "n.s." means no statistical difference, p >0.05; ". Times." indicates significant differences, p <0.05 is 0.0001;
FIG. 11 is a wrinkle removing effect chart (VC 98 USB photo) of the effective example 1;
FIG. 12 is a wrinkle removal effect chart (VC 98 USB photo) of working example 2;
FIG. 13 is a view showing the effect of relieving redness in working example 1 (Visia-CR Cross Polarized Light Red Areas mode photograph);
FIG. 14 is a view showing the effect of relieving redness in working example 2 (visual-CR Cross Polarized Light Red Areas mode photograph).
Detailed Description
For a better understanding of the present invention, the following examples are set forth to illustrate the present invention further, but are not to be construed as limiting its scope.
The preparation method of the brown alga microzyme fermentation lysate used in the embodiment comprises the following steps:
1) Soaking brown algae, cleaning, pulverizing into 100 mesh, and sterilizing with plasma beam with irradiation intensity of 6000kGy for 40min;
2) Mixing 1) with 3 times of purified water, and inoculating 2×10 of baker's yeast suspension 5 Fermenting cfu/ml at 36 ℃ for 72 hours to obtain fermentation liquor;
3) Breaking cell wall of thallus in the fermentation liquor by a high-pressure homogenizer, obtaining brown algae microzyme fermentation crude extract, filtering by 400 meshes to obtain brown algae microzyme fermentation lysate, and regulating the dry matter content (solid matter content after drying) to about 10%.
The preparation method of the lactobacillus/pear juice fermentation filtrate used in the embodiment comprises the following steps: inoculating lactobacillus 1×10 into pear juice culture medium 6 Performing fermentation culture on cfu/ml, wherein the fermentation temperature is 37 ℃ and the fermentation time is 48 hours; and (3) centrifuging at 8000r/min after fermentation, and taking supernatant as lactobacillus/pear juice fermentation filtrate. The pear juice culture medium is as follows: cleaning pear, squeezing with a juicer (no additional water is added in the squeezing process) to obtain freshly squeezed pear juice, adding 20% of freshly squeezed pear juice into MRS culture medium, and sterilizing to obtain pear juice culture medium.
Example 1: preparation of the emulsion
1.0g of sucrose laurate, 4.0g of butanediol and 1.5g of hexanediol are heated and mixed, 1.94g of purified water is added, and the temperature is reduced to 38 ℃ to form liquid I; mixing 2.0g of white pool flower seed oil, 2.0g of marlura oil and 2.5g of caprylic/capric triglyceride to form a liquid II; adding 2.0g of brown algae microzyme fermentation lysate (0.2 g based on dry matter), 0.06g of calcium propionate and 3.0g of lactobacillus/pear juice fermentation liquor into the mixed liquor of the liquor I, and uniformly stirring and mixing to form liquor III; slowly adding the second liquid into the third liquid, and homogenizing at 10000r/min for 8min to obtain uniform emulsion.
Example 2: preparation of the emulsion
1.2g of sucrose laurate, 3.0g of pentanediol and 2.5g of glycerol are heated and mixed, 1.45g of purified water is added, and the temperature is reduced to 38 ℃ to form liquid I; mixing 2.0g camellia seed oil, 2.0g marlura oil and 3.0g caprylic/capric triglyceride to form a liquid two; adding 2.0g of brown algae microzyme fermentation lysate (0.2 g based on dry matter), 0.05g of calcium propionate and 2.8g of lactobacillus/pear juice fermentation liquor into the mixed liquor of the liquor I, and uniformly stirring and mixing to form liquor III; slowly adding the second liquid into the third liquid, and homogenizing at 10000r/min for 8min to obtain uniform emulsion.
Example 3: preparation of the emulsion
1.5g of sucrose laurate, 0.3g of octanediol and 4.0g of glycerol are heated and mixed, 2.93g of purified water is added, and the temperature is reduced to 38 ℃ to form liquid I; mixing 2.0g camellia seed oil, 2.0g marlura oil and 3.0g caprylic/capric triglyceride to form a liquid two; adding 2.2g of brown algae microzyme fermentation lysate (0.22 g based on dry matter), 0.07g of calcium propionate and 2.0g of lactobacillus/pear juice fermentation liquor into the mixed liquor of the liquor I, and uniformly stirring and mixing to form liquor III; slowly adding the second liquid into the third liquid, and homogenizing at 10000r/min for 8min to obtain uniform emulsion.
Control group 1: phaeophyta yeast fermentation lysate group
The same procedure as in example 1 was repeated except that 5.0g of the brown algae yeast fermentation lysate (0.5 g on a dry matter basis) was used and that lactobacillus/pear juice fermentation broth was not used, to obtain a brown algae yeast fermentation lysate group (emulsion).
Control group 2: lactobacillus/pear juice fermentation filtrate group
Using 5.0g of lactobacillus/pear juice fermentation broth, the lysate was fermented without using brown algae yeast, and the rest of components, amounts and preparation method were the same as in example 1, to obtain lactobacillus/pear juice fermentation filtrate group (emulsion).
The emulsions of example 1 and comparative examples 1-2 were used in the following test examples, in which the above three emulsions were diluted to 5. Mu.g/mL with DMEM/F12 (1:1) medium, respectively, for the following test examples 2 and 3.
Test example 1: detection and analysis of skin physiological parameters and skin microbial flora of different aging people
A healthy female 262 aged 18-65 years was selected, wrinkle detection was performed by a Visia-CR facial image analyzer, and three groups (W0, W1, and W2 respectively: W0: no wrinkles; W1: slight wrinkles; W2: more wrinkles and deep: three groups of volunteers after classification were 134, 105 and 23 persons, respectively) were classified according to the wrinkle condition. The following skin physiological parameter detection and skin microbial flora detection were performed on the above three persons.
Skin physiological parameter detection: usingCM 825 skin moisture tester, < >>TM 300 skin moisture loss tester, < >>SM 815 skin oil tester, cutometer dual MPA580 skin elasticity tester, respectively, measured skin moisture content, skin percutaneous water loss rate (TEWL), sebum amount, elasticity and firmness of corresponding areas under cheekbones of left and right sides of the volunteer. The detection results are shown in FIG. 1.
Detection of skin microbial flora: 3 sterile cotton swab swabs are dipped in sterile physiological saline (0.9% physiological sodium chloride solution-0.1% Tween-20), repeatedly rubbed for at least 15 times at the symmetrical positions of the left and right sides of the face of a volunteer and the middle three positions of the forehead, and the cotton swab swabs are rotated during the rubbing, and the sampling area is 3cm multiplied by 3cm. After the collection was completed, 16S rDNA sequencing was performed and by analysis, the differences in skin microbial flora diversity (Chao index (a) and Shannon index) and community flora composition (genus level) were seen for the different wrinkle groups. The analysis results are shown in fig. 2 and 3.
The test found that propionibacterium was most abundant in the facial skin flora of the tested population and showed a positive correlation with sebum levels. As wrinkles increase, the facial skin microbial flora and flora are more diverse, with the small wrinkles and wrinkles being more abundant and the deep group propionibacteria (Cutibacterium) being significantly less abundant than the no wrinkles group (see fig. 3).
Test example 2: effect of lotions on skin tissue morphology
EX vivo skin tissue cultured in vitro was used as subject, and divided into 4 groups of 3 replicates, and UVR irradiation was performed on 4 groups (wherein UVA was 30J/cm) 2 ，UVB 50mJ/cm 2 For 4 days after irradiation), the UVR irradiation was not treated to give NC group, and the UVR irradiation was followed by treatment with brown algae yeast fermentation lysate (5. Mu.g/mL), lactobacillus/pear juice fermentation filtrate (5. Mu.g/mL), and the emulsion of the present invention (5. Mu.g/mL) to give 3 experimental groups, each of which was administered at 2. Mu.L/cm to the surface of EX vivo skin tissue 2 Then by observing the tissue H&E dyeing to evaluate the effect of the emulsion.
The test results are shown in fig. 4, and compared with the NC control group, the brown algae yeast fermentation lysate, the lactobacillus/pear juice fermentation filtrate and the emulsion group can obviously thicken the epidermis living cells, obviously increase the density of dermis fibroblast and reduce vacuoles; and the emulsion group has better promoting effect than single component of the brown alga microzyme fermentation lysate and lactobacillus/pear juice fermentation filtrate.
Test example 3: effect of emulsions on elastin synthesis
EX vivo skin tissue cultured in vitro was used as subject, and divided into 4 groups of 3 replicates, and UVR irradiation was performed on 4 groups (wherein UVA was 30J/cm) 2 ，UVB50mJ/cm 2 4 days of irradiation), N without any treatment after UVR irradiationGroup C, 3 experimental groups were treated with UVR-irradiated brown algae yeast fermentation lysate (5. Mu.g/mL), lactobacillus/pear juice fermentation filtrate (5. Mu.g/mL), and the emulsion of the present invention (5. Mu.g/mL), and 2. Mu.L/cm of each of the EX vivo skin tissue surfaces was administered 2 The effect of the emulsion was evaluated by detecting the elastin content in the tissue.
As shown in the test results in FIG. 5, compared with the NC control group, both the brown algae yeast fermentation lysate and the emulsion group can promote the increase of the elastin content (increase of brown color area after immunohistochemical staining) in EX vivo skin tissues, and the emulsion group has better promoting effect than the single components of the brown algae yeast fermentation lysate and the lactobacillus/pear juice fermentation filtrate.
Test example 4: skin micro-ecology improvement of aged skin by emulsion
Preparing 0.4wt% of acrylic copolymer aqueous solution, adding 20g of emulsion prepared in example 1 into 85.0g of acrylic copolymer aqueous solution, and stirring and mixing uniformly to form an essence product.
The beneficial effect of the essence containing the emulsion on skin micro-ecology is evaluated by taking 30-40 years old Chinese healthy female population as a target and taking 33 people as a total, continuously using the essence containing the emulsion for 7 days in the morning and evening after a tester cleans the face every day, and measuring indexes such as changes of skin flora before and after the volunteer product is used.
The results show that: the use of the essence containing the emulsion, the increase in propionibacterium abundance in the population tested (figures 6-7, table 1), gradually transformed the skin microbiota with signs of aging to a younger skin microbiota composition.
TABLE 1 comparison of the relative abundance of skin bacterial species before and after use of test products (genus level)
Test example 5: skin physiological state change of emulsion on aging skin
Preparing 0.4wt% of acrylic copolymer aqueous solution, adding 20g of emulsion prepared in example 1 into 85.0g of acrylic copolymer aqueous solution, and stirring and mixing uniformly to form an essence product.
The total number of people is 33 by taking 30-40 years old Chinese healthy female population as the object, so that testers can clean the face every day, and can continuously use the essence containing the emulsion for 28 days and 56 days in the morning and evening, and then respectively detect the face. The beneficial effect of the emulsion-containing essence on the skin physiology was evaluated by measuring the change in skin physiology before and after use of the volunteer product. A photograph of the subject's face is taken and parameters such as skin wrinkles, dermis layer echo intensity, fine line length, fine line count, subcutaneous red area color a value, etc. are measured. The data results before and after the product is used and the evaluation results are compared by a statistical test method to judge whether the statistical difference exists.
The test results are shown in fig. 8-10, and the results show that the skin wrinkles SEw are obviously reduced after 28 days of use, which indicates that the tested product has the wrinkle removing effect after 28 days of use; after 56 days of use, the skin wrinkles SEw, the fine line score, the fine line length, the fine line count and the subcutaneous red area color a value are all significantly reduced, while the dermis layer echo intensity is significantly increased, which indicates that the tested product has the effects of removing wrinkles, resisting aging, relieving and the like after 56 days of use.
2 testers with obvious wrinkle-removing and anti-aging effects after 28 days of using the product are selected, and effective examples are shown in figures 11-12. 2 test subjects with obvious red relieving effect after 28 days of using the product were selected, and effective examples are shown in fig. 13 and 14. As can be seen from the figures: the effect is remarkable.
Claims (6)
1. An emulsion with effect of improving skin microecology and skin physiological state of aging is characterized by comprising at least three effective components of brown algae microzyme fermentation lysate, lactobacillus/pear juice fermentation filtrate and short-chain fatty acid salt;
the dosage of the three active ingredients in the emulsion is as follows: each 20g of the emulsion contains 0.1-0.3g of brown algae saccharomycete fermentation lysate, 0.5-8.0g of lactobacillus/pear juice fermentation filtrate and 0.05-0.1g of short-chain fatty acid salt, wherein the dosage of the brown algae saccharomycete fermentation lysate is calculated by dry matter;
the brown algae microzyme fermentation lysate is as follows: inoculating brown algae water solution into saccharomycetes for fermentation, breaking walls of fermentation liquor by a high-pressure homogenizer, crushing thalli, and filtering by 200-400 meshes to obtain the brown algae water solution;
the lactobacillus/pear juice fermentation filtrate is as follows: fermenting and culturing lactobacillus in pear juice culture medium, centrifuging, and collecting supernatant to obtain pear juice culture medium, wherein 20+ -5% of freshly squeezed pear juice is added into MRS culture medium;
the short-chain fatty acid salt is one or two of lactate, butyrate, propionate and isobutyrate.
2. An emulsion for improving the microecological and physiological conditions of aged skin according to claim 1, wherein the three active ingredients are used in the emulsion in the following amounts: each 20g of emulsion contains 0.2g of brown algae microzyme fermented lysate, 3.0g of lactobacillus/pear juice fermented filtrate and 0.06g of short-chain fatty acid salt.
3. The emulsion for improving the microecological and physiological states of aged skin according to claim 1 or 2, wherein sucrose ester emulsifier, natural oil, caprylic/capric triglyceride, polyalcohol and water are added to prepare the emulsion, and each 20g of the emulsion contains 0.1-0.3g of brown algae yeast fermentation lysate, 0.5-8.0g of lactobacillus/pear juice fermentation filtrate, 0.05-0.1g of short chain fatty acid salt, 0.2-2.0g of sucrose ester emulsifier, 0.5-10.0g of natural oil, 1.0-4.0g of caprylic/capric triglyceride, 0.5-10.0g of polyalcohol and the balance of water.
4. An emulsion having an improving effect on the micro-ecology of aged skin and the physiological state of skin as claimed in claim 3,
the sucrose ester emulsifier is one or two of sucrose laurate, sucrose palmitate and sucrose stearate;
the natural oil is one or two of Beacon flower seed oil, mala oil, camellia seed oil, coconut oil and bitter almond oil;
the polyalcohol is one or two of octanediol, pentanediol, butanediol, glycerol, dipropylene glycol and hexanediol.
5. A method of preparing an emulsion according to claim 3, comprising the steps of:
s1: mixing sucrose ester emulsifier with polyalcohol, heating for dissolving, adding water, and cooling to form liquid I;
s2: mixing natural oil and caprylic/capric triglyceride to form liquid II;
s3: adding the brown algae microzyme fermented lysate, short-chain fatty acid salt and lactobacillus/pear juice fermentation liquor into the mixed liquor of the first liquid, and uniformly stirring and mixing to form a third liquid;
s4: slowly adding the second liquid into the third liquid, and homogenizing at high speed to form uniform emulsion.
6. Use of the emulsion according to claim 1 or 2 for the preparation of a skin care product for improving the micro-ecology and physiological state of aged skin by adding to an anti-ageing skin care product which is micro-ecology friendly to skin.
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