CN105853341B - A kind of anti-aging skin care product and preparation method thereof containing epidermal stem cells secretin - Google Patents
A kind of anti-aging skin care product and preparation method thereof containing epidermal stem cells secretin Download PDFInfo
- Publication number
- CN105853341B CN105853341B CN201610316104.2A CN201610316104A CN105853341B CN 105853341 B CN105853341 B CN 105853341B CN 201610316104 A CN201610316104 A CN 201610316104A CN 105853341 B CN105853341 B CN 105853341B
- Authority
- CN
- China
- Prior art keywords
- epidermal stem
- stem cells
- skin care
- preparation
- plant extracts
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Classifications
-
- 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
- A61K8/99—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
-
- 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
-
- 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
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Birds (AREA)
- Epidemiology (AREA)
- Dermatology (AREA)
- Gerontology & Geriatric Medicine (AREA)
- Botany (AREA)
- Microbiology (AREA)
- Mycology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Cosmetics (AREA)
Abstract
The invention belongs to skin care fields, and in particular to a kind of anti-aging skin care product and preparation method thereof containing epidermal stem cells secretin.The skin care item are grouped as by the group of following weight percent meter：Epidermal stem cells secretin 0.000001-0.0005%, plant extracts 2~8%, saualane 1~2%, EDTA-2Na 0.10~0.25%, glycerine 5~10%, polyglutamic acid 0.05~1%,
Description
Technical field
The invention belongs to skin care fields, and in particular to a kind of anti-aging skin care product containing epidermal stem cells secretin and
Preparation method.
Background technology
Skin aging is the skin aging phenomenon as caused by natural cause or non-natural factor, show as wrinkle appearance,
The relaxation of skin, scrambling pigmentation, blood vessel dilatation, keratinization of epidermis is bad, abnormality proliferation occur, corium elastomer becomes
Property and catabolite accumulation etc..Skin aging is closely related with cell ageing, especially with fibroblast and keratinocyte
Aging be closely connected.The secretion of fibroblast collagen during aging is reduced, while the induction energy of cell glue protoenzyme
Power raises, and collagen is caused to decompose enhancing, and to promote skin elasticity to decline, wrinkle occurs.In addition, keratinocyte exists
Skin moisturizing, moisturizing, barrier and skin and formation etc. are many-sided to have very important adjustment effect.Keratinocyte declines
It is slow with the relevant gene expression of skin repair in old process, and damage related gene expression and increase, to promote skin old
Change.
Stem cell is a kind of seed cell of not yet differentiation and development, has unlimited self-renewing replication capacity, and can break up
At specific organization, this is stem cell for improving looks or even the basis of regeneration.Studies have shown that in the animal body of aging
Interior, epidermal cell accretion rate slows down, therefore skin speckle increases, and wrinkle increases.The major function of epidermal stem cells is supplement
Epidermal cell maintains the balance of Skin Cell and repairs damaged tissues.Promote the Proliferation, Differentiation of epidermal stem cells that can make skin
Keep vigorous metabolism.In addition, the self-replacation of stem cell and pluripotent differentiation also rely on the association of a variety of growth factors
Same facilitation, studies have shown that stem cell can secrete some active materials in incubation, including growth factor, cell because
The substances such as son, active peptides, protein, growth hormone, trace element, promote itself amplification and differentiation daughter cell increasing
It grows.
Currently, someone promotes cell-secretion factor (also known as stem cell secretion element) from the raffinate that stem cell is cultivated and is used for
Cosmetics.Tai lifts " whole world head branch cell cosmetics will be listed in Korea Spro " one delivered in Science and Technology Daily and the article pointed out Korea Spro in 2011
FCB Pharmicell companies of state have developed the makeup of human stem cell culture solution by core material of stem cell culture solution
Product, the said firm is in experiments it is found that the collagen amount generated in the skin fibroblasts of injection stem cell culture solution is
5 times or more of control group.Chinese patent application 201110452259.6 discloses a kind of compound life for beautifying skin reparation
Object preparation and preparation method, the composite biological agent contain cord blood stem cell active constituent and placenta active constituent, have and promote
Into the metabolism of puerpera's epidermal stem cells, to realize beauty reparation, the purpose of plaque dispelling skin care, but the composite biological agent is
Be prepared according to conventional skin care item production method, i.e., by cord blood stem cell active matter, placenta active matter in proportion with
White class cosmetics etc. are made according to conventional skin care item production method in the inert matter mixing of conventional cosmetic.It is well known that dry thin
Active constituent in born of the same parents' beauty product is easy to inactivate at normal temperatures, to keep its activity, must just have and be made under specified conditions
Standby and preservation.Therefore, the effect of such stem cell beauty product remains to be discussed.In addition, the stem cell extract application of heterologous
The adverse reactions such as allergic reaction are easily caused in people.
Invention content
Effectively skin can repair and what will not be caused allergic reaction contain the purpose of the present invention is to provide a kind of
There is the anti-aging skin care product and preparation method thereof of epidermal stem cells secretin.The anti-aging skin care product active constituent can be preferably
Ground preserves, and promotes skin hyperplasia ability to give full play to it, improves skin elasticity, improve the effect of skin quality, there is no heterologous
Property etc. caused by reasons phenomena such as allergic reaction, and preparation method is simple, and property is stablized.
The present invention is by following technical solution to realize above-mentioned purpose：
A kind of anti-aging skin care product containing epidermal stem cells secretin, which is characterized in that the skin care item are by following heavy
The group of amount percentages is grouped as：Epidermal stem cells secretin 0.000001-0.0005%, plant extracts 2~8%, spiny dogfish
Alkane 1~2%, EDTA-2Na 0.10~0.25%, glycerine 5~10%, polyglutamic acid 0.05~1%,K350 0.06~
0.12%, isopropyl myristate 2~6%, cetostearyl alcohol 0.5~2%, dimethyl silicone polymer 1~2%, surplus are water.
Preferably, the anti-aging skin care product, which is characterized in that the skin care item by following weight percent meter group
It is grouped as：It is epidermal stem cells secretin 0.000125%, plant extracts 6%, saualane 1%, EDTA-2Na 0.15%, sweet
Oil 8%, polyglutamic acid 0.50%,K350 0.10%, isopropyl myristate 4%, cetostearyl alcohol 1%, poly- diformazan
Radical siloxane 2%, surplus are water.
Further, the epidermal stem cells secretin is the mixing of the various active factor of the epidermal stem cells secretion of people
Object.
Further, the plant extracts is prepared by the raw material of following weight percentage：Circassian bean 16~28%,
Sherry flower 14~30%, wilsonii 12~28%, purpleflower holly bark 14~26% and raspberry 10~22%.
Preferably, the plant extracts is prepared by the raw material of following weight percentage：Circassian bean 22%, sherry flower
22%, wilsonii 20%, purpleflower holly bark 20% and raspberry 16%.
Further, the preparation method of the epidermal stem cells secretin includes the following steps：
(1) acquisition of human epidermal stem cell
It takes operation to cut off or scrape oral epithelium tissue, tissue is transferred to big mould containing the celebrating of penicillin, streptomysin and sulfuric acid
In the D-PBSA (phosphate buffer) of element, laboratory is transported back under the conditions of dry ice or liquid nitrogen frozen.It rinses 2-3 times, is added repeatedly
37 DEG C of digestion 30min of trypsase, oral epithelium tissue is shredded, epithelial cell is filtered with 70-100um cell filters, receives
Collect filtrate.5min is centrifuged under the conditions of 1200r/min, abandons supernatant, is softly blown with Defined Keratinocyte-SFM culture mediums
It beats, obtains cell suspension, human epidermal stem cell is isolated and purified using collagen rapid attachment method；
(2) culture of human epidermal stem cell
The human epidermal stem cell that step (1) obtains is seeded in containing 100U/ml penicillin and 100ug/ml streptomysins
In Defined Keratinocyte-SFM culture mediums, 37 DEG C, 5%CO2Incubator is incubated 12-24h, abandons culture medium, adds again
Enter it is fresh contain penicillin and streptomysin Defined Keratinocyte-SFM culture mediums, cultivate 1-3d, abandon culture medium, add
Enter Defined Keratinocyte-SFM culture mediums to continue to cultivate.When human epidermal stem cell close to when 90% fusion, passage is trained
It supports；
(3) preparation of epidermal stem cells secretin
Human epidermal stem cell is expanded into 1-5 generations, collects culture supernatant, by 0.22 μm of sterilizing filter mistake of supernatant
Filter merges the filtrate per a generation up to epidermal stem cells secretin.
Further, the preparation method of the plant extracts includes the following steps：
It takes ingredients in proportion respectively, washes away impurity, crushed after being dried crosses 50~60 mesh sieve, merges coarse powder, medicine is added
The ethyl alcohol that 8~12 times of amount volume fractions of material coarse powder total weight are 50~70%, Microwave Extraction 4~6 minutes, microwave power 400
~500W, Extracting temperature are 40~60 DEG C, and extracting solution is filtered, and extraction 1 time is repeated by similarity condition, merge extracting solution, decompression
It is concentrated into the medicinal extract for surveying that relative density is 1.05~1.15 at 60 DEG C, plant extracts is made.
In addition, the present invention also provides the preparation methods of the anti-aging skin care product comprising following steps：
Step 1：Epidermal stem cells secretin, excipient, water for injection are uniformly mixed, vacuum freeze drying is carried out, obtains
Freeze-dried powder is spare；
Step 2：By EDTA-2Na, polyglutamic acid and water mixing, 80 DEG C, 6~10min of homogeneous are heated to, stirring to dissolving
Completely, material A is obtained；
Step 3：By glycerine, saualane, isopropyl myristate, cetostearyl alcohol and dimethyl silicone polymer mixing, add
For heat to 80~90 DEG C, 10~15min of homogeneous, stirring is complete to dissolving, and obtains material B；It is slowly added to material B into material A, stirs
Uniformly, start to cool down；
Step 4：When temperature is down to 30 DEG C or so, to emulsification pot in be added plant extracts,K350 and step 1
Freeze-dried powder obtained, stirs evenly, and discharges to plate；
Wherein, the excipient described in step 1 is mannitol and L-arginine with 1: 1 proportioning mixture obtained；
Wherein, the vacuum freeze drying step of step 1 is：Epidermal stem cells secretin, excipient, water for injection are mixed
It is uniformly placed in -70 DEG C of freeze dryers, is cooled to -50 DEG C hereinafter, pre-freeze 6h, vacuumizes 8h, is to slowly warm up to -20 DEG C, then with 5
DEG C/speed of h is warming up to 25 DEG C, heat preservation and dryness 2h to get.
Compared with prior art, advantage of the invention is that：
(1) active constituent of anti-aging skin care product provided by the invention be epidermal stem cells secretin and plant extracts,
The epidermal stem cells secretin is the mixture for the various active factor that the epidermal stem cells of people are secreted, therefore can avoid dry
Immunological rejection caused by cell extract heterologous reduces anaphylactoid incidence；In addition, having in plant extracts
Imitating ingredient has promotion Transdermal absorption effect, and the active constituent of anti-aging skin care product is made to be deep into dermal layer of the skin and subcutaneous tissue
The effect of giving full play to reparation skin, promoting skin hyperplasia ability；In addition, the active ingredient in plant extracts also has albumen
Enzyme inhibition is hydrolyzed, to delay the bioactive ingredients in epidermal stem cells secretin thin in dermal cell and subcutaneous tissue
Intracellular improves epidermal stem cells secretin senile-resistant efficacy by the speed of proteolytic degradation.
(2) present invention is during preparing anti-aging skin care product, using Vacuum Freezing & Drying Technology epidermal stem cells
Freeze-dried powder is made in secretin, is conducive to preserve the bioactive ingredients in epidermal stem cells secretin, anti-to preferably play
Aging effect.
(3) anti-aging skin care product provided by the invention can significantly improve skin aging, reduce the generation of adverse reaction, increase
Water content of stratum corneum and fat content, effect is better than the more vigor lotions of the long-drawn-out Lay of Shiseido, and preparation method is simple, and property is steady
It is fixed, it is suitble to industrialized production.
Specific implementation mode
The present invention is further described below by way of specific implementation mode, but the present invention is not limited only to following embodiment.
Embodiment 1
Anti-aging skin care product described in the embodiment of the present invention 1 is grouped as by the group of following weight percent meter：Epidermal stem is thin
Intracrine element 0.000125%, plant extracts 6%, saualane 1%, EDTA-2Na 0.15%, glycerine 8%, polyglutamic acid
0.50%,K350 0.10%, isopropyl myristate 4%, cetostearyl alcohol 1%, dimethyl silicone polymer 2%, it is remaining
Amount is water；The epidermal stem cells secretin is the mixture for the various active factor that the epidermal stem cells of people are secreted；The plant
Object extract is prepared by the raw material of following weight percentage：Circassian bean 22%, sherry flower 22%, wilsonii 20%, Chinese ilex
Skin 20% and raspberry 16%.
Preparation method：
(1) preparation of epidermal stem cells secretin：
The acquisition of human epidermal stem cell：Take operation cut off or scraping oral epithelium tissue, by tissue be transferred to containing penicillin,
In the D-PBSA (phosphate buffer) of streptomysin and gentamicin sulphate, laboratory is transported back under the conditions of dry ice or liquid nitrogen frozen.
It rinses 3 times repeatedly, 37 DEG C of digestion 30min of trypsase is added, oral epithelium tissue are shredded, with 100um cell filter mistakes
Epithelial cell is filtered, filtrate is collected.5min is centrifuged under the conditions of 1200r/min, abandons supernatant, with Defined Keratinocyte-SFM
Culture medium is softly blown and beaten, and cell suspension is obtained, and human epidermal stem cell is isolated and purified using collagen rapid attachment method；
The culture of human epidermal stem cell：Human epidermal stem cell is seeded in containing 100U/ml penicillin and 100ug/ml chains
In mycin Defined Keratinocyte-SFM culture mediums, 37 DEG C, 5%CO2Incubator is incubated for 24 hours, abandons culture medium, is added again
Enter it is fresh contain penicillin and streptomysin Defined Keratinocyte-SFM culture mediums, cultivate 2d, abandon culture medium, be added
Defined Keratinocyte-SFM culture mediums continue to cultivate.When human epidermal stem cell is merged close to 90%, secondary culture；
The preparation of epidermal stem cells secretin：Human epidermal stem cell was expanded into for 5 generations, culture supernatant is collected, by supernatant
It is filtered with 0.22 μm of sterilizing filter, merges the filtrate per a generation up to epidermal stem cells secretin；
(2) preparation of plant extracts：
It takes ingredients in proportion respectively, washes away impurity, crushed after being dried crosses 60 mesh sieve, merges coarse powder, it is thick that medicinal material is added
The ethyl alcohol that 10 times of amount volume fractions of powder total weight are 60%, Microwave Extraction 5 minutes, microwave power 450W, Extracting temperature 45
DEG C, extracting solution is filtered, extraction 1 time is repeated by similarity condition, merges extracting solution, being concentrated under reduced pressure into survey relative density at 60 DEG C is
Plant extracts is made in 1.05 medicinal extract；
(3) preparation of anti-aging skin care product：
Step 1：Epidermal stem cells secretin, excipient, water for injection are uniformly mixed and are placed in -70 DEG C of freeze dryers,
- 50 DEG C are cooled to hereinafter, pre-freeze 6h, vacuumizes 8h, -20 DEG C is to slowly warm up to, then 25 DEG C are warming up to the speed of 5 DEG C/h, protects
The dry 2h of temperature is spare to get freeze-dried powder；
Step 2：By EDTA-2Na, polyglutamic acid and water mixing, 80 DEG C, 6~10min of homogeneous are heated to, stirring to dissolving
Completely, material A is obtained；
Step 3：By glycerine, saualane, isopropyl myristate, cetostearyl alcohol and dimethyl silicone polymer mixing, add
For heat to 80~90 DEG C, 10~15min of homogeneous, stirring is complete to dissolving, and obtains material B；It is slowly added to material B into material A, stirs
Uniformly, start to cool down；
Step 4：When temperature is down to 30 DEG C or so, to emulsification pot in be added plant extracts,K350 and step 1
Freeze-dried powder obtained, stirs evenly, and discharges to plate；
Wherein, the excipient described in step 1 is mannitol and L-arginine with 1: 1 proportioning mixture obtained.
Embodiment 2
Anti-aging skin care product described in the embodiment of the present invention 2 is grouped as by the group of following weight percent meter：Epidermal stem is thin
Intracrine element 0.0005%, plant extracts 8%, saualane 2%, EDTA-2Na 0.25%, glycerine 10%, polyglutamic acid
1%,K350 0.12%, isopropyl myristate 6%, cetostearyl alcohol 2%, dimethyl silicone polymer 2%, surplus are
Water；The epidermal stem cells secretin is the mixture for the various active factor that the epidermal stem cells of people are secreted；The plant carries
Object is taken to be prepared by the raw material of following weight percentage：Circassian bean 16%, sherry flower 14%, wilsonii 28%, purpleflower holly bark
20% and raspberry 22%.
The preparation method is the same as that of Example 1.
Embodiment 3
Anti-aging skin care product described in the embodiment of the present invention 3 is grouped as by the group of following weight percent meter：Epidermal stem is thin
Intracrine element 0.000001%, plant extracts 2%, saualane 1%, EDTA-2Na 0.10%, glycerine 5%, polyglutamic acid
0.05%,K350 0.06%, isopropyl myristate 2%, cetostearyl alcohol 0.5%, dimethyl silicone polymer 1%,
Surplus is water；The epidermal stem cells secretin is the mixture for the various active factor that the epidermal stem cells of people are secreted；It is described
Plant extracts is prepared by the raw material of following weight percentage：Circassian bean 28%, sherry flower 30%, wilsonii 12%, winter
Green peel 20% and raspberry 10%.
The preparation method is the same as that of Example 1.
Embodiment 4
Anti-aging skin care product described in the embodiment of the present invention 4 is grouped as by the group of following weight percent meter：Epidermal stem is thin
Intracrine element 0.00025%, plant extracts 6%, saualane 1%, EDTA-2Na 0.15%, glycerine 8%, polyglutamic acid
0.25%,K350 0.10%, isopropyl myristate 4%, cetostearyl alcohol 1%, dimethyl silicone polymer 2%, it is remaining
Amount is water；The epidermal stem cells secretin is the mixture for the various active factor that the epidermal stem cells of people are secreted；The plant
Object extract is prepared by the raw material of following weight percentage：Circassian bean 20%, sherry flower 20%, wilsonii 20%, Chinese ilex
Skin 26% and raspberry 14%.
The preparation method is the same as that of Example 1.
Embodiment 5
Anti-aging skin care product described in the embodiment of the present invention 5 is grouped as by the group of following weight percent meter：Epidermal stem is thin
Intracrine element 0.00005%, plant extracts 8%, saualane 2%, EDTA-2Na 0.25%, glycerine 10%, polyglutamic acid
0.75%,K350 0.12%, isopropyl myristate 6%, cetostearyl alcohol 2%, dimethyl silicone polymer 2%, it is remaining
Amount is water；The epidermal stem cells secretin is the mixture for the various active factor that the epidermal stem cells of people are secreted；The plant
Object extract is prepared by the raw material of following weight percentage：Circassian bean 28%, sherry flower 30%, wilsonii 12%, Chinese ilex
14% and raspberry 16%.
The preparation method is the same as that of Example 1.
Comparative example 1
Anti-aging skin care product described in comparative example 1 of the present invention is grouped as by the group of following weight percent meter：Epidermal stem is thin
Intracrine element 0.000125%, saualane 1%, EDTA-2Na 0.15%, glycerine 8%, polyglutamic acid 0.50%,K350
0.10%, isopropyl myristate 4%, cetostearyl alcohol 1%, dimethyl silicone polymer 2%, surplus are water；The epidermal stem
Cell secretin is the mixture for the various active factor that the epidermal stem cells of people are secreted.(lack plant compared with Example 1 to carry
Take object this component)
Preparation method reference implementation example 1.
Comparative example 2
Anti-aging skin care product described in comparative example 2 of the present invention is grouped as by the group of following weight percent meter：Plant extract
Object 6%, saualane 1%, EDTA-2Na 0.15%, glycerine 8%, polyglutamic acid 0.50%,K350 0.10%, Pork and beans
Cool isopropyl propionate 4%, cetostearyl alcohol 1%, dimethyl silicone polymer 2%, surplus are water；The plant extracts is by following heavy
The raw material of amount percentage meter is prepared：Circassian bean 22%, sherry flower 22%, wilsonii 20%, purpleflower holly bark 20% and raspberry
16%.(this component of shortage epidermal stem cells secretin compared with Example 1)
Preparation method reference implementation example 1.
Comparative example 3
Anti-aging skin care product described in comparative example 3 of the present invention is grouped as by the group of following weight percent meter：Epidermal stem is thin
Intracrine element 0.000125%, plant extracts 6%, saualane 1%, EDTA-2Na 0.15%, glycerine 8%, polyglutamic acid
0.50%,K350 0.10%, isopropyl myristate 4%, cetostearyl alcohol 1%, dimethyl silicone polymer 2%, it is remaining
Amount is water；The epidermal stem cells secretin is the mixture for the various active factor that the epidermal stem cells of people are secreted；The plant
Object extract is prepared by the raw material of following weight percentage：Sherry flower 30%, wilsonii 28%, purpleflower holly bark 20% and cover basin
Son 22%.(this component of plant extracts shortage circassian bean compared with Example 1)
Preparation method reference implementation example 1.
Test example one, anti-aging skin care product resisting age of skin clinical observation of the present invention
1. test method
The tested volunteer of women that aging occur in 540 skins of screening, age 28-50 Sui, average age 35 years old are selected in mark
It is accurate：Dry skin, elasticity reduce, and the skin ageing phenomenons such as wrinkle, pigmentation, the colour of skin be gloomy occur；During entire experiment
It is ready to avoid solarization, or face uses the sunscreen product of SPF >=30 when contact sunlight；It is anti-without using other during entire experiment
Aging product.
540 tested volunteers of women are randomly divided into 9 groups, respectively control group, embodiment 1-5 groups, comparative example 1-3 groups,
Every group 60, wherein control group subject uses Shiseido Urara Multy-Vitalizing Emulsion (Shiseido
The more vigor lotions of long-drawn-out Lay), early, after late face cleaning suitable lotion is taken to be applied to face；Embodiment 1-5 groups and comparative example 1-3 groups by
Examination person uses anti-aging skin care product made from 1-5 of the embodiment of the present invention, comparative example 1-3 respectively, and suitable shield is taken after early, late face cleaning
Skin product are applied to face, and the entire test period of each group is 90d, the adverse reaction occurred during record experiment, and to subject into
Row effect assessment.
Skin aging is assessed：(glabella line, forehead line, fish are evaluated by the wrinkle in 5 tested areas of observation subject
Tail line, nasolabial groove, lip week wrinkle), judgment criteria：There is no wrinkle；Slightly：There are the shallow wrinkle of 2-3 items, length < 1.5cm；Moderate：
There are the shallow wrinkle of 2-6 items, length < 3cm；Severe：There are several main wrinkles, length to reach 4cm, while with shallow wrinkle.
Safety evaluation：There is slight discomfort, erythema, drying, itch and shouting pain in subject after primary evaluation uses product
A situation arises for equal adverse reactions.
Water content, the fat content for detecting recipient site using non-invasive instrument to subject in 90d, carry out statistical
Analysis.
2. test result
Table 1 after the test each group subject skin aging assessment
| Group | There is no wrinkle (number of cases) | Slightly (number of cases) | Moderate (number of cases) | Severe (number of cases) |
| Control group | 38 | 16 | 4 | 2 |
| Embodiment 1 | 46 | 12 | 2 | 0 |
| Embodiment 2 | 43 | 14 | 3 | 0 |
| Embodiment 3 | 42 | 14 | 4 | 0 |
| Embodiment 4 | 41 | 15 | 4 | 0 |
| Embodiment 5 | 41 | 14 | 5 | 0 |
| Comparative example 1 | 32 | 19 | 5 | 4 |
| Comparative example 2 | 30 | 20 | 5 | 5 |
| Comparative example 3 | 34 | 17 | 6 | 3 |
The adverse reaction evaluation of each group subject during table 2 is tested
| Group | Without (number of cases) | Slight discomfort (number of cases) | Erythema (number of cases) | Dry (number of cases) | Itch (number of cases) | Shouting pain (number of cases) |
| Control group | 45 | 7 | 2 | 2 | 1 | 3 |
| Embodiment 1 | 55 | 3 | 0 | 2 | 0 | 0 |
| Embodiment 2 | 52 | 5 | 0 | 2 | 1 | 0 |
| Embodiment 3 | 51 | 5 | 0 | 3 | 1 | 0 |
| Embodiment 4 | 49 | 6 | 0 | 4 | 1 | 0 |
| Embodiment 5 | 48 | 7 | 0 | 4 | 1 | 0 |
| Comparative example 1 | 39 | 9 | 3 | 5 | 2 | 2 |
| Comparative example 2 | 40 | 8 | 2 | 4 | 3 | 3 |
| Comparative example 3 | 42 | 9 | 3 | 2 | 2 | 2 |
Water content of stratum corneum during the experiment of 3 each group subject of table and fat content evaluation
Note：Compared with 0d,*P < 0.05,**P < 0.01.
Shiseido Urara Multy-Vitalizing Emulsion are the more vigor lotions of the Japanese long-drawn-out Lay of Shiseido, are
Anti-aging product of new generation containing epidermal growth factor aims at the product of Chinese women preparation, and it comprises triple amino acid
Ingredient is repaired, moisturizing, brilliant white, Cucumber Extract promote the synthesis of collagen；The drying of skin, low and deep and wrinkle can be effectively improved
The problems such as line, while protecting skin, obstructs extraneous harmful substance, makes skin youth, brilliant white, gloss and flexible；The table contained
Skin growth factor active constituent, enables that skin is smooth, compacts.
This research swings the more vigor lotions of Lay as a control group with Japanese Shiseido, evaluates the effect of anti-aging skin care product of the present invention
Fruit, the results showed that, the anti-aging skin care product prepared using 1-5 of the embodiment of the present invention, no subjects skin's aging reaches serious
Degree, and control group has 2；Simultaneously subject do not occur yet erythema, shouting pain adverse reaction, and control group have 2 it is tested
There is erythema in person, and shouting pain occur in 3 subjects；In addition, the anti-aging skin care product prepared using 1-5 of the embodiment of the present invention can be shown
Write and increase subjects skin's water content of stratum corneum and fat content, wherein the anti-aging skin care product effect prepared with embodiment 1 compared with
It is good, increase subjects skin's water content of stratum corneum and reaches pole significant difference, the above result shows that, 1-5 systems of the embodiment of the present invention
Standby anti-aging skin care product can significantly improve skin aging, reduce the generation of adverse reaction, increase water content of stratum corneum and grease
Content, and with the anti-aging skin care product best results of embodiment 1, and it is better than control group.Anti-aging skin care product of the present invention can be effective
Increase keratoderma water content, replenishing water and preserving moisture, and by increase sebum, play the role of greasy skin.1 He of comparative example
2 lack plant extracts and epidermal stem cells secretin respectively, improve the decrease with obvious effects of skin aging, adverse reaction
Incidence obviously increases, and increases subjects skin's water content of stratum corneum and fat content effect is not notable, shows that the present invention is anti-ageing
Both active ingredient plant extracts and epidermal stem cells secretin in old skin care item have mutually coordinated, the work mutually promoted
With lacking one can cause the effect of anti-aging to reduce.Active ingredient in plant extracts, which has, promotes Transdermal absorption to make
With making that the active constituent of anti-aging skin care product is deep into dermal layer of the skin and subcutaneous tissue gives full play to reparation skin, promote skin
The effect of skin Reproductive activity；Epidermal stem cells secretin is containing there are many on the influential growth factor of Skin Cell activity, cell
The various actives substance such as the factor, active peptides, trace element, amino acid, may advantageously facilitate skin metabolism, slow down aging.
The effect of circassian bean has dispelling wind and heat, and eliminating dampness is antipruritic, moisturizing beauty treatment, and there is stronger inhibiting effect to proteolytic enzyme,
Plant extracts in comparative example 3 lacks circassian bean this component, and the effect of its anti-aging is made slightly to be better than comparative example 1 and comparative example
2, but with the obvious decrease of the effect of embodiment 1-5, show being effectively split into point by inhibiting proteolytic enzyme in circassian bean
Activity delay epidermal stem cells secretin metabolic breakdown in the cell, give full play to epidermal stem cells secretin promote skin it is new
Old metabolism, the effect of anti-aging.The above result shows that each ingredient collocation is reasonable in anti-aging skin care product prescription of the present invention, phase
Mutual gain has significant effect to improving skin aging, and adverse reaction is low, safe.
It the above is only the preferred embodiment of the present invention, it is noted that above-mentioned preferred embodiment is not construed as pair
The limitation of the present invention, protection scope of the present invention should be subject to claim limited range.For the art
For those of ordinary skill, without departing from the spirit and scope of the present invention, several improvements and modifications can also be made, these change
Protection scope of the present invention is also should be regarded as into retouching.
Claims (6)
1. a kind of anti-aging skin care product containing epidermal stem cells secretin, which is characterized in that the skin care item are by following weight
The group of percentages is grouped as：Epidermal stem cells secretin 0.000001-0.0005%, plant extracts 2~8%, saualane 1
~2%, EDTA-2Na 0.10~0.25%, glycerine 5~10%, polyglutamic acid 0.05~1%,K350 0.06~
0.12%, isopropyl myristate 2~6%, cetostearyl alcohol 0.5~2%, dimethyl silicone polymer 1~2%, surplus are water；
The epidermal stem cells secretin is the mixture for the various active factor that the epidermal stem cells of people are secreted；
The preparation method of the epidermal stem cells secretin includes the following steps：
(1) acquisition of human epidermal stem cell
It takes operation to cut off or scrape oral epithelium tissue, tissue is transferred to containing penicillin, streptomysin and gentamicin sulphate
In D-PBSA (phosphate buffer), laboratory being transported under the conditions of dry ice or liquid nitrogen frozen back, being rinsed 2-3 times repeatedly, pancreas egg is added
37 DEG C of digestion 30min of white enzyme, oral epithelium tissue is shredded, epithelial cell is filtered with 70-100um cell filters, collects filter
Liquid centrifuges 5min under the conditions of 1200r/min, abandons supernatant, softly blown and beaten, obtained with Defined Keratinocyte-SFM culture mediums
Cell suspension isolates and purifies human epidermal stem cell using collagen rapid attachment method；
(2) culture of human epidermal stem cell
The human epidermal stem cell that step (1) obtains is seeded in containing 100U/ml penicillin and 100ug/ml streptomysins Defined
In Keratinocyte-SFM culture mediums, 37 DEG C, 5%CO2Incubator is incubated 12-24h, abandons culture medium, rejoins fresh
Containing penicillin and streptomysin Defined Keratinocyte-SFM culture mediums, 1-3d is cultivated, culture medium is abandoned, is added
Defined Keratinocyte-SFM culture mediums continue to cultivate, when human epidermal stem cell is merged close to 90%, secondary culture；
(3) preparation of epidermal stem cells secretin
Human epidermal stem cell is expanded into 1-5 generations, collects culture supernatant, supernatant is filtered with 0.22 μm of sterilizing filter,
Merge the filtrate per a generation up to epidermal stem cells secretin；
The plant extracts is prepared by the raw material of following weight percentage：Circassian bean 16~28%, sherry flower 14~
30%, wilsonii 12~28%, purpleflower holly bark 14~26% and raspberry 10~22%；
The preparation method of the plant extracts includes the following steps：It takes ingredients in proportion respectively, impurity is washed away, after dry
It crushes, crosses 50~60 mesh sieve, merge coarse powder, the second that 8~12 times of amount volume fractions of medicinal material coarse powder total weight are 50~70% is added
Alcohol, Microwave Extraction 4~6 minutes, microwave power are 400~500W, and Extracting temperature is 40~60 DEG C, and extracting solution is filtered, by same
Batten part repeats extraction 1 time, merges extracting solution, is concentrated under reduced pressure into the medicinal extract for surveying that relative density is 1.05~1.15 at 60 DEG C, system
Obtain plant extracts.
2. anti-aging skin care product according to claim 1, which is characterized in that the skin care item are by following weight percent meter
Group be grouped as：Epidermal stem cells secretin 0.000125%, plant extracts 6%, saualane 1%, EDTA-2Na
0.15%, glycerine 8%, polyglutamic acid 0.50%,K350 0.10%, isopropyl myristate 4%, cetostearyl alcohol
1%, dimethyl silicone polymer 2%, surplus are water.
3. anti-aging skin care product according to claim 1 or 2, which is characterized in that the plant extracts is by following weight
The raw material of percentage meter is prepared：Circassian bean 22%, sherry flower 22%, wilsonii 20%, purpleflower holly bark 20% and raspberry 16%.
4. a kind of preparation method of anti-aging skin care product as described in claim 1, which is characterized in that include the following steps：
Step 1：Epidermal stem cells secretin, excipient, water for injection are uniformly mixed, vacuum freeze drying is carried out, must be lyophilized
Powder is spare；
Step 2：By EDTA-2Na, polyglutamic acid and water mixing, 80 DEG C, 6~10min of homogeneous are heated to, stirring is complete to dissolving,
Obtain material A；
Step 3：By glycerine, saualane, isopropyl myristate, cetostearyl alcohol and dimethyl silicone polymer mixing, it is heated to
80~90 DEG C, 10~15min of homogeneous, stirring is complete to dissolving, and obtains material B；Material B is slowly added into material A, stirring is equal
It is even, start to cool down；
Step 4：When temperature is down to 30 DEG C or so, to emulsification pot in be added plant extracts,K350 and step 1 are made
Freeze-dried powder, stir evenly, to plate discharge.
5. the preparation method of anti-aging skin care product according to claim 4, which is characterized in that the excipient described in step 1
It is mannitol and L-arginine with 1:1 proportioning mixture obtained.
6. the preparation method of anti-aging skin care product according to claim 4, which is characterized in that the vacuum refrigeration of step 1 is dry
Dry step is：Epidermal stem cells secretin, excipient, water for injection are uniformly mixed and are placed in -70 DEG C of freeze dryers, it is cooling
To -50 DEG C hereinafter, pre-freeze 6h, vacuumizes 8h, -20 DEG C are to slowly warm up to, then 25 DEG C are warming up to the speed of 5 DEG C/h, heat preservation is dry
Dry 2h to get.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610316104.2A CN105853341B (en) | 2016-05-11 | 2016-05-11 | A kind of anti-aging skin care product and preparation method thereof containing epidermal stem cells secretin |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610316104.2A CN105853341B (en) | 2016-05-11 | 2016-05-11 | A kind of anti-aging skin care product and preparation method thereof containing epidermal stem cells secretin |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN105853341A CN105853341A (en) | 2016-08-17 |
| CN105853341B true CN105853341B (en) | 2018-10-26 |
Family
ID=56631834
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610316104.2A Active CN105853341B (en) | 2016-05-11 | 2016-05-11 | A kind of anti-aging skin care product and preparation method thereof containing epidermal stem cells secretin |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105853341B (en) |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108078909A (en) * | 2018-01-12 | 2018-05-29 | 广州赛莱拉干细胞科技股份有限公司 | Stem cell toner |
| CN108078908A (en) * | 2018-01-12 | 2018-05-29 | 广州赛莱拉干细胞科技股份有限公司 | Spirulina extract stem cell smoothing toner |
| CN110548001A (en) * | 2019-09-06 | 2019-12-10 | 沈阳细胞治疗工程技术研发中心有限公司 | Repair anti-aging skin care product containing umbilical cord mesenchymal stem cell exosomes |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2013209302A (en) * | 2012-03-30 | 2013-10-10 | Kose Corp | Agent containing extract of young apple fruits for maintenance of skin stem cells |
| CN103142447B (en) * | 2013-03-18 | 2015-01-14 | 山西医科大学 | Epidermal stem cell active component and cosmetic preparation containing same |
| CN105267102A (en) * | 2015-11-18 | 2016-01-27 | 中山市艾丽意素生物科技有限公司 | Natural herbal anti-aging face cream and method for preparing same |
-
2016
- 2016-05-11 CN CN201610316104.2A patent/CN105853341B/en active Active
Also Published As
| Publication number | Publication date |
|---|---|
| CN105853341A (en) | 2016-08-17 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105106028B (en) | A kind of peptide composition for hair growth | |
| KR20140008428A (en) | Skin cream | |
| CN108186548A (en) | A kind of preparation method of the stem cell factor Essence with anti-aging effects | |
| CN107854418A (en) | A kind of human umbilical cord mesenchymal stem cells excretion external use creams and preparation method thereof | |
| CN110339147A (en) | A kind of composition for external application and application thereof | |
| KR101897401B1 (en) | Cosmetic composition for skin desquamation comprising powder of captured spicule using ultra high pressure as effective component | |
| JP2003510364A (en) | Method for producing a composition by extraction of nacres, comprising all components of nacre, composition obtained using this method, and use of this composition in cosmetics and dermatology | |
| JP2003002813A (en) | Skin care composition | |
| CN104644461A (en) | Application of compound with effect of promoting synthetic activity of collagen in anti-aging cosmetics | |
| CN105853341B (en) | A kind of anti-aging skin care product and preparation method thereof containing epidermal stem cells secretin | |
| CN110123729A (en) | A kind of jelly film and preparation method thereof | |
| KR20180089373A (en) | A cosmetic care method for improving cosmetic effect. | |
| CN114344188A (en) | Hair loss prevention and hair growth promoting composition and preparation method thereof | |
| CN112807417B (en) | Collagen-containing skin-whitening and anti-wrinkle medicine or cosmetic | |
| CN111419760A (en) | Fresh black tiger leaf extracting solution and preparation method and application thereof | |
| KR102358611B1 (en) | Cosmetic composition for enhancing skin elasticity or improving skin wrinkle containing Polygonum Cuspidatum root exracts | |
| KR20100096447A (en) | Cosmetic composition comprising matrials cultured adult stem cells derived from swine placenta tissue and proteins extracted therefrom | |
| JP5642354B2 (en) | Use of Brassocatrea marcella Koss orchid extract as an active agent to prevent or delay the appearance of signs of skin aging | |
| CN105147575A (en) | Hair restorer containing stem cells and preparation method thereof | |
| KR101521268B1 (en) | Manufacturing method of fermented composition of amniotic fluid of Sturgeon, fermented composition of amniotic fluid of Sturgeon obtained therefrom and cosmetic composition comprising the same | |
| CN108721202B (en) | Roselle stem cell freeze-dried powder eye cream and preparation method thereof | |
| CN111135118A (en) | Anti-aging polypeptide composition | |
| KR100903283B1 (en) | Hair growing material which contains a deer antler ingredient and its manufacturing method | |
| KR102151501B1 (en) | Composition for improving hair and head skin | |
| KR102047442B1 (en) | Composite for mask pack |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20230719 Address after: Building 102, Building 1, No. 76, Xieshi Road, Shibi Street, Shibi Village, Panyu District, Guangzhou City, Guangdong Province, 511496 Patentee after: Guangdong Kewei Zhili Biopharmaceutical Co.,Ltd. Address before: Room 301, 401, 39 BAOYING Avenue, Guangzhou Free Trade Zone, Guangdong 510730 Patentee before: GUANGDONG COOWAY BIOTECHNOLOGY Co.,Ltd. |
|
| TR01 | Transfer of patent right |