CN112826786A - Targeted scar repairing mesh construction composition and application thereof - Google Patents
Targeted scar repairing mesh construction composition and application thereof Download PDFInfo
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- CN112826786A CN112826786A CN202011623748.9A CN202011623748A CN112826786A CN 112826786 A CN112826786 A CN 112826786A CN 202011623748 A CN202011623748 A CN 202011623748A CN 112826786 A CN112826786 A CN 112826786A
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- A—HUMAN NECESSITIES
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- 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
- A61K8/96—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]
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- A—HUMAN NECESSITIES
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- 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
- A61Q19/06—Preparations for care of the skin for countering cellulitis
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- 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
- A61K2800/74—Biological properties of particular ingredients
- A61K2800/78—Enzyme modulators, e.g. Enzyme agonists
- A61K2800/782—Enzyme inhibitors; Enzyme antagonists
Abstract
The invention discloses a composition for constructing a target repair scar tissue network, which comprises an India swertia extract, a sweet orange fruit extract, a blue-green microalgae extract and an avocado fruit extract. The effect of elasticity removal of striae gravidarum of the skin is improved by scientific proportioning of the plant active extract composition, the India swertia extract has the effects of resisting inflammation, inhibiting bacteria, increasing the thickness of epidermis and the like, the sweet orange fruit extract has the effects of resisting wrinkles and oxidation and promoting the renewal of keratinocytes, the collagen stimulators are actively screened from the natural components in the blue-green microalgae extract to attack the regeneration of collagen and improve the water content of the skin, and the avocado fruit extract has good skin affinity, is easily absorbed by the skin, has strong absorption on ultraviolet rays and has the effect of lipolysis and firming. The composition for constructing the target repair scar tissue mesh has the effects of enhancing skin toughness, lightening wrinkles, resisting wrinkles, tightening and eliminating fat and the like.
Description
Technical Field
The invention relates to the technical field of cosmetics, in particular to a composition for constructing a target repairing scar tissue mesh and application thereof.
Background
Stretch marks refer to the phenomenon of skin fiber rupture of the human body during pregnancy, are one of stretch marks, are white or silvery glossy scar lines, mainly appear on the abdominal wall, and also can appear on the inner and outer sides of thighs, buttocks and the like, and are particularly obvious for the primiparous woman. Striae gravidarum is mainly affected by hormones in pregnancy, and in addition, the abdominal distension causes the elastic fibers and collagen fibers of the skin to be damaged, so that striae gravidarum appears. After delivery, striae atrophicae transform into striae gravidarum. Once it appears, it does not disappear and is accompanied by skin laxity, breast drop, abdominal fat accumulation, which seriously affects the postpartum physical and mental health of women.
Striae gravidarum is light and heavy, and the following methods are common at present: 1. the treatment is carried out by means of laser, the principle is that the laser with specific wavelength and pulse width is emitted to selectively penetrate through epidermis to stimulate an injured collagen layer to generate a new collagen tissue, the new collagen tissue grows at a normal speed to gradually repair the injured collagen layer and improve the texture and the state of the skin, but the method can only be properly lightened, so that the new collagen tissue is not obvious and can not be completely removed, and some postoperative repair treatment is needed after the operation, otherwise, the effect which is proper to the contrary can be achieved; 2. the abdominal skin-pulling operation, namely the abdominal wall plastic operation, recovers the elasticity and the compactness of the abdominal fascia through re-suturing the abdominal fascia, improves the abdominal tension, and can push the contents originally belonging to the abdominal cavity inwards to achieve the purpose of removing the striae gravidarum.
Disclosure of Invention
In order to solve the above technical problems, the first aspect of the present invention provides a targeted scar repairing web construction composition comprising an India swertia extract, a sweet orange fruit extract, a blue-green microalgae extract, and an avocado fruit extract.
As a preferred technical scheme, the weight ratio of the India swertia extract, the sweet orange fruit extract, the blue-green microalgae extract and the avocado fruit extract is (40-70): (25-45): (12-30): (15-40).
As a preferred technical scheme, the weight ratio of the India swertia extract, the sweet orange fruit extract, the blue-green microalgae extract and the avocado fruit extract is (45-65): (30-40): (15-25): (20-35).
As a preferred technical scheme, the weight ratio of the India swertia extract, the sweet orange fruit extract, the blue-green microalgae extract and the avocado fruit extract is (50-60): (32-38): (18-22): (22-32).
As a preferred technical scheme, the preparation method of the India swertia extract comprises the following steps:
a. callus culture: taking tender leaves of India swertia as explants to carry out plant tissue culture, and collecting the growing callus blocks;
b. seed cell culture: separating the callus blocks by an enzymolysis method, filtering, separating, collecting seed cells, and inoculating the seed cells to a culture medium for culture;
c. preparing an extract: adding ethanol into cultured cells, extracting to obtain crude extract, removing solvent, and purifying with chromatographic column.
As a preferred technical solution, the preparation method of the sweet orange fruit extract comprises the following steps: cleaning fructus Citri sinensis, slicing, oven drying, pulverizing to obtain powder, adding water, freezing, thawing under microwave heating, ultrasonic vibrating, centrifuging to obtain supernatant, filtering, concentrating, eluting with snake cage resin, removing solvent, freeze drying, and grinding.
As a preferred technical solution, the preparation method of the blue-green microalgae extract comprises the following steps: soaking the blue-green microalgae with 80-95 v% ethanol for 24 hours, collecting a primary ethanol extract, soaking the blue-green microalgae with 70-80 v% ethanol for 24 hours, collecting a secondary ethanol extract, combining the two ethanol extracts, removing ethanol, freeze-drying the remainder to obtain a freeze-dried substance, grinding and sieving to obtain the product.
As a preferred technical scheme, the preparation method of the avocado fruit extract comprises the following steps: removing kernels and peels of avocado fruits, drying and crushing the avocado fruits, adding a solvent, carrying out ultrasonic treatment for 3-4 hours, removing the solvent, freeze-drying, and grinding to obtain the avocado fruit extract.
In a second aspect of the invention there is provided the use of a targeted scar tissue mesh building composition as described above for the prevention or repair of stretch marks.
As a preferred technical scheme, the composition is used for preparing gel, cream, emulsion and essential oil.
Has the advantages that: the invention provides a construction composition for targeted scar repairing netting, which is characterized in that the effect of elasticity removal of striae gravidarum of skin is improved by using the scientific proportion of plant active extract compositions such as India swertia extract, sweet orange fruit extract, blue-green microalgae extract and avocado fruit extract, the India swertia extract has the effects of resisting inflammation, inhibiting bacteria, increasing the thickness of epidermis and the like, the sweet orange fruit extract has the effects of resisting wrinkles, resisting oxidation and promoting the renewal of keratinocytes, collagen stimulators are actively screened from natural components in the blue-green microalgae extract to attack the regeneration of collagen, the water content of the skin is improved, and the avocado fruit extract has good affinity to the skin of a human body, is very easy to be absorbed by the skin, has strong absorptivity to ultraviolet rays and has the effects of lipolysis and firming. The composition for constructing the target repair scar tissue mesh has the effects of enhancing skin toughness, lightening wrinkles, resisting wrinkles, tightening and eliminating fat and the like.
Drawings
To further illustrate the beneficial effects of an anti-HPV viral gel and a method for preparing the same provided in the present invention, the accompanying drawings are provided, and it should be noted that the drawings provided in the present invention are only selected as individual examples in all drawings and are not intended to limit the claims, and all other corresponding maps obtained by the drawings provided in the present application should be considered as being within the protection scope of the present application.
FIG. 1 illustrates the mechanism of action of the present invention.
Fig. 2 is a result of efficacy test for epidermal regeneration in example 1 of the present invention.
FIG. 3 shows the results of in vitro tests on cell energy and cell activity in example 1 of the present invention.
FIG. 4 is the results of the in vivo test for keratinocyte renewal of example 1 of the present invention.
FIG. 5 shows the results of in vivo tests on dermal cells in example 1 of the present invention.
Detailed Description
The disclosure may be understood more readily by reference to the following detailed description of preferred embodiments of the invention and the examples included therein. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. In case of conflict, the present specification, including definitions, will control.
The term "prepared from …" as used herein is synonymous with "comprising". The terms "comprises," "comprising," "includes," "including," "has," "having," "contains," "containing," or any other variation thereof, as used herein, are intended to cover a non-exclusive inclusion. For example, a composition, process, method, article, or apparatus that comprises a list of elements is not necessarily limited to only those elements but may include other elements not expressly listed or inherent to such composition, process, method, article, or apparatus.
The conjunction "consisting of …" excludes any unspecified elements, steps or components. If used in a claim, the phrase is intended to claim as closed, meaning that it does not contain materials other than those described, except for the conventional impurities associated therewith. When the phrase "consisting of …" appears in a clause of the subject matter of the claims rather than immediately after the subject matter, it defines only the elements described in the clause; other elements are not excluded from the claims as a whole.
When an amount, concentration, or other value or parameter is expressed as a range, preferred range, or as a range of upper preferable values and lower preferable values, this is to be understood as specifically disclosing all ranges formed from any pair of any upper range limit or preferred value and any lower range limit or preferred value, regardless of whether ranges are separately disclosed. For example, when a range of "1 to 5" is disclosed, the described range should be interpreted to include the ranges "1 to 4", "1 to 3", "1 to 2 and 4 to 5", "1 to 3 and 5", and the like. When a range of values is described herein, unless otherwise stated, the range is intended to include the endpoints thereof and all integers and fractions within the range.
The singular forms "a", "an" and "the" include plural referents unless the context clearly dictates otherwise. "optional" or "any" means that the subsequently described event or events may or may not occur, and that the description includes instances where the event occurs and instances where it does not.
Approximating language, as used herein throughout the specification and claims, is intended to modify a quantity, such that the invention is not limited to the specific quantity, but includes portions that are literally received for modification without substantial change in the basic function to which the invention is related. Accordingly, the use of "about" to modify a numerical value means that the invention is not limited to the precise value. In some instances, the approximating language may correspond to the precision of an instrument for measuring the value. In the present description and claims, range limitations may be combined and/or interchanged, including all sub-ranges contained therein if not otherwise stated.
In addition, the indefinite articles "a" and "an" preceding an element or component of the invention are not intended to limit the number requirement (i.e., the number of occurrences) of the element or component. Thus, "a" or "an" should be read to include one or at least one, and the singular form of an element or component also includes the plural unless the stated number clearly indicates that the singular form is intended.
In order to solve the above technical problems, the first aspect of the present invention provides a targeted scar repairing web construction composition comprising an India swertia extract, a sweet orange fruit extract, a blue-green microalgae extract, and an avocado fruit extract.
Swertia pseudochinensis, known as Swertian bimaculata (Sieb. etZucc.) hook. f. et Thoms. ex C.B.Clark, is also known as swertia herb, bitter in taste and cold in nature. The swertia japonica Makino contains isovitexin, isoorientin, 1, 3-dihydroxy-4, 5-dimethoxyxanthone (1, 3-dihydroxy-4, 5-dimethoxyxanthone), 1, 3-dihydroxy-4, 5-dimethoxyxanthone-1-O-beta-D-glucopyranoside (1, 3-dihydroxy-4, 5-dimethoxyxanthone-l-O-beta-D-glucopyranoside), 1, 3-dihydroxy-4, 5-dimethoxyxanthone-3-O-beta-D-glucopyranoside (1, 3-dihydroxy-4, 5-dimethoxyxanthone-3-O-beta-D-glucopyranoside), swertisin (swertisin), isoorientin (swertisin), Swertisin (swero-side), etc.
In some preferred embodiments, the method for preparing the India swertia extract comprises the steps of:
a. callus culture: taking tender leaves of India swertia as explants to carry out plant tissue culture, and collecting the growing callus blocks;
b. seed cell culture: separating the callus blocks by an enzymolysis method, filtering, separating, collecting seed cells, and inoculating the seed cells to a culture medium for culture;
c. preparing an extract: adding ethanol into cultured cells, extracting to obtain crude extract, removing solvent, and purifying with chromatographic column.
In some further preferred embodiments, the method of preparing the India swertia extract comprises the steps of:
a. callus culture: taking tender leaves of India swertia as explants to carry out plant tissue culture on an MS solid culture medium, and collecting the growing callus blocks;
b. seed cell culture: taking 2-8 g of callus blocks, adding 120-130 mL of 0.04-0.12 g/L pectinase solution to treat the callus blocks, filtering, performing centrifugal separation, collecting seed cells, inoculating the seed cells to an MS liquid culture medium for culture, performing amplification culture after the culture time is 5-10 days, performing amplification culture in a fermentation tank for 10-20 days, and performing centrifugal collection on the cultured cells;
c. preparing an extract: taking 40-60 g of cultured cells, extracting for 2 times by using 50-80 v% ethanol, combining extracting solutions, filtering to obtain a crude extract, removing the ethanol to obtain an extract, mixing the extract with silica gel according to a weight ratio of 1: (3-5), mixing, and filling into a chromatographic column, wherein the volume ratio is 10: 1, eluting with an ethyl acetate/ethanol mixture, and then eluting with a solvent with a volume ratio of 8: 1, and finally eluting with an ethyl acetate/ethanol mixture in a volume ratio of 2: eluting with ethyl acetate/ethanol mixed solution of 1 to obtain purified eluent, and removing solvent to obtain the final product.
The MS medium in the present invention is Murashige and Skoog medium well known to those skilled in the art, and the method for obtaining it may be commercially available.
Sweet orange fruit, known by the academia Citrus sinensis (L.) Osbeck, is sweet, pungent, slightly bitter and slightly warm in nature. The fruit contains flavonoid glycoside, lactone, alkaloid, organic acid, etc. The flavonoid glycosides include hesperidin (heperidin), narirutin (narirutin), isosakuranetin-7-rutinoside (isosakuranetin-7-rutinoside), naringenin-4 '-glucoside-7-rutinoside (naringenin-4' -glucoside-7-rutinoside), naringin (naringin), 3-beta-D-glucoside (limonin-3-beta-D-glucoside) and O-D-xylosylvitexin (O-D-xylosylvitexin); the lactone contains a bifactone bitter component limonin, namely phellodendron lactone (phellodendron) and a derivative limonin (narcotine); the organic acid mainly comprises citric acid (citric acid) and malic acid (malic acid), and also comprises phloroglucinol-beta-D-glucoside (phloroglucinol-beta-D-glucoside, phyllorin), saccharides, vitamins, calcium, phosphorus, iron, etc.
In some preferred embodiments, the method for preparing the sweet orange fruit extract comprises the following steps: cleaning fructus Citri sinensis, slicing, oven drying, pulverizing to obtain powder, adding water, freezing, thawing under microwave heating, ultrasonic vibrating, centrifuging to obtain supernatant, filtering, concentrating, eluting with snake cage resin, removing solvent, freeze drying, and grinding.
In some further preferred embodiments, the method of preparing the sweet orange fruit extract comprises the steps of: cleaning, slicing, drying and crushing sweet orange fruits to obtain powder, adding 12 times of water, placing the powder in an ultra-low temperature refrigerator, freezing at the temperature of-80 to-60 ℃, heating and unfreezing for 3min by microwave at the temperature of 60-80 ℃, ultrasonically shaking the unfreezing liquid for 0.5-2 h, centrifuging to obtain supernatant, filtering, concentrating, passing through a snake cage resin column, washing the resin column with water, eluting with 70 v% ethanol to obtain eluent, removing the solvent, freeze-drying and grinding to obtain the sweet orange fruit extract.
Microalgae refers to a micro group of algae whose morphology can be identified under a microscope, and the microalgae cultured or produced in large quantities by applying biotechnology belong to 4 phyla: the blue-green microalgae extract is the extract of microalgae of cyanophyta and chlorophyta. Microalgae cells contain high-value nutritional components such as proteins, lipids, algal polysaccharides, beta-carotene, various inorganic elements (such as Cu, Fe, Se, Mn, Zn and the like) and the like. The protein content of microalgae is very high, and the microalgae is an important source of single-cell protein; the vitamin A, the vitamin E, the thiamine, the riboflavin, the pyridoxine, the vitamin B12, the vitamin C, the biotin, the inositol, the folic acid, the calcium pantothenate, the nicotinic acid and the like contained in the microalgae increase the value of the microalgae as single-cell protein; the alga has higher carotenoid content, has the functions of coloring and nutrition, can be used for preventing and treating cancers, resisting radiation, delaying senility, enhancing the immunity of organisms and other physiological functions, is chemically synthesized into trans-beta-carotene, has the functions of carcinogenesis and teratogenesis on human bodies, has higher functions of resisting cancers and cardiovascular diseases than all-trans-isomer, and has the beta-carotene content in the alga powder as high as 14 percent; the algae cell has high content of glycerol, is a high-quality cosmetic raw material, and is an organic intermediate with wide application in chemical industry, light industry and pharmaceutical industry; the algal polysaccharide complex can be used as an immunologic adjuvant to enhance antigenicity and immunologic function of organisms, obviously inhibit solid tumors and play a role in resisting tumors.
In some preferred embodiments, the method of preparing the blue-green microalgae extract comprises the steps of: soaking the blue-green microalgae with 80-95 v% ethanol for 24 hours, collecting a primary ethanol extract, soaking the blue-green microalgae with 70-80 v% ethanol for 24 hours, collecting a secondary ethanol extract, combining the two ethanol extracts, removing ethanol, freeze-drying the remainder to obtain a freeze-dried substance, grinding and sieving to obtain the product.
In some further preferred embodiments, the method of preparing the blue-green microalgae extract comprises the steps of: soaking the blue-green microalgae with 80-95 v% ethanol in an amount which is 8 times that of the blue-green microalgae for 24 hours, collecting primary ethanol extract, soaking the blue-green microalgae with 70-80 v% ethanol in an amount which is 5 times that of the blue-green microalgae for 24 hours, collecting secondary ethanol extract, combining the two ethanol extract, removing ethanol, freeze-drying residues to obtain freeze-dried substances, grinding and sieving to obtain the product.
Avocado fruit, known under the name perseamerica mill, is a fruit with high nutritive value, contains various vitamins, rich fat and protein, has high contents of sodium, potassium, magnesium, calcium and the like, and can be eaten as raw fruit and also can be eaten as dish and can; the nut contains fatty oil, is non-drying oil, and has mild fragrance.
In some preferred embodiments, the method of preparing the avocado fruit extract comprises the steps of: removing kernels and peels of avocado fruits, drying and crushing the avocado fruits, adding a solvent, carrying out ultrasonic treatment for 3-4 hours, removing the solvent, freeze-drying, and grinding to obtain the avocado fruit extract.
In some further preferred embodiments, the method of preparing the avocado fruit extract comprises the steps of: removing kernels and peels of avocado fruits, drying and crushing the pulp, adding 80-95 v% ethanol with the amount being 10 times of that of the pulp, carrying out ultrasonic treatment for 3-4 hours, removing the ethanol, carrying out freeze drying on the remainder to obtain a freeze-dried substance, grinding and sieving to obtain the avocado fruit.
The preparation method of the composition for constructing the target repair scar tissue network provided by the invention can be physical blending.
In some preferred embodiments, the weight ratio of the India swertia extract, the sweet orange fruit extract, the blue-green microalgae extract and the avocado fruit extract is (40-70): (25-45): (12-30): (15-40); further preferably, the weight ratio of the India swertia extract, the sweet orange fruit extract, the blue-green microalgae extract and the avocado fruit extract is (45-65): (30-40): (15-25): (20-35); furthermore, the weight ratio of the India swertia extract, the sweet orange fruit extract, the blue-green microalgae extract and the avocado fruit extract is (50-60): (32-38): (18-22): (22-32).
The inventor finds that the scientific proportion of the plant active extract compositions such as India swertia extract, sweet orange fruit extract, blue-green microalgae extract, avocado fruit extract and the like is utilized to increase the elasticity of the skin and remove the striae gravidarum, and the India swertia extract has the effects of resisting inflammation, inhibiting bacteria, increasing the thickness of the epidermis and the like; the fructus Citri sinensis extract has effects of reducing wrinkle, resisting oxidation, and promoting keratinocyte regeneration; actively screening collagen stimulators from natural components in the blue-green microalgae extract, attacking collagen regeneration and improving the water content of skin; the avocado fruit extract has good skin affinity, can be easily absorbed by skin, has strong absorption to ultraviolet, and has lipolysis and firming effects. The composition for constructing the target repair scar tissue mesh has the effects of enhancing skin toughness, lightening wrinkles, resisting wrinkles, tightening and eliminating fat and the like.
As shown in fig. 1, the main action mechanism of the composition for targeted scar tissue repair provided by the invention is as follows: accelerating keratinocyte proliferation, enhancing epidermal cell regeneration, strengthening epidermal structure and increasing epidermal thickness; strengthening the immune system of the skin, resisting inflammation and relieving; inducing the neogenesis of collagen, fibronectin, elastin; effective in resisting oxidation, scavenging free radicals, protecting Langerhans cells, and increasing barrier protection ability; effectively reducing the release of elastase inhibitors and protecting the growth of elastin; promoting the regeneration and reconstruction of dermis and recovering the vitality of the young cells; tightening skin elasticity, eliminating subcutaneous adipose tissue, and reducing orange peel wrinkles; preventing and eliminating striae gravidarum and revitalizing skin; moisten skin, and endow skin with high moisture retention and moisture.
The inventor finds in research that a plurality of target action sites exist in the process of repairing stretch marks: 1. acting on keratinocytes to promote the production of keratinocyte growth factor and increase the thickness of epidermis; promoting the synthesis of TRPV3 channel protein, opening a corresponding cation channel, introducing calcium ions into keratinocytes, inducing the synthesis of vesicle protein, and accelerating the renewal and exfoliation of the keratinocytes; avoid dehydration-induced epidermal toxicity, increase skin water content; 2. acting on epidermis layer, scavenging free radicals, protecting Langerhans cells, improving the activity of resisting microorganisms, and enhancing the oxidation resistance of epidermis and the natural defense of skin; 3. acting on dermis layer to provide energy for cells, promote synthesis of elastase inhibitor (SKALP), reduce activity of leukocyte elastase (HLE), and increase collagen, fibronectin, and elastin in dermis layer; 4. acting on adipose tissue, promoting synthesis of ZAG protein (zinc-alpha-2 glycoprotein), increasing cAMP (cyclic adenosine monophosphate) ratio, decomposing cellulite, and tightening skin.
Examples
The present invention will be described in detail with reference to specific examples. The following examples will assist those skilled in the art in further understanding the invention, but are not intended to limit the invention in any way. It should be noted that variations and modifications can be made by persons skilled in the art without departing from the spirit of the invention. All falling within the scope of the present invention.
Example 1
Example 1 provides a composition for targeted scar repair construction, comprising 55 parts by weight of India swertia extract, 35 parts by weight of sweet orange fruit extract, 20 parts by weight of blue-green microalgae extract, and 27 parts by weight of avocado fruit extract.
The preparation method of the India swertia extract comprises the following steps:
a. callus culture: taking tender leaves of India swertia as explants to carry out plant tissue culture on an MS solid culture medium, and collecting the growing callus blocks;
b. seed cell culture: taking 5g of callus blocks, adding 125mL of 0.1g/L pectinase solution to treat the callus blocks, filtering, centrifugally separating, collecting seed cells, inoculating the seed cells to an MS liquid culture medium for culture, culturing at room temperature for 7 days, then carrying out amplification culture, carrying out amplification culture in a fermentation tank, culturing for 15 days, and centrifugally collecting the cultured cells;
c. preparing an extract: taking 50g of cultured cells, extracting for 2 times by using 70 v% ethanol, combining extracting solutions, filtering to obtain a crude extract, removing the ethanol to obtain an extract, mixing the extract with silica gel according to a weight ratio of 1: 4 mixing, loading into a chromatographic column, and mixing by using a mixture prepared by mixing the components in a volume ratio of 10: 1, eluting with an ethyl acetate/ethanol mixture, and then eluting with a solvent with a volume ratio of 8: 1, and finally eluting with an ethyl acetate/ethanol mixture in a volume ratio of 2: eluting with ethyl acetate/ethanol mixed solution of 1 to obtain purified eluent, and removing solvent to obtain the final product.
The preparation method of the sweet orange fruit extract comprises the following steps: cleaning sweet orange fruits, slicing, drying, pulverizing to obtain powder, adding 12 times of water, placing in an ultra-low temperature refrigerator, freezing at-70 deg.C, thawing for 3min under microwave heating at 70 deg.C, thawing the frozen solution for 1h under ultrasonic oscillation, centrifuging to obtain supernatant, filtering, concentrating, passing through a column with snake cage resin, washing the resin column with water, eluting with 70 v% ethanol to obtain eluate, removing solvent, freeze drying, and grinding.
The preparation method of the blue-green microalgae extract comprises the following steps: soaking blue-green microalgae with 85 v% ethanol 8 times of the amount of the ethanol for 24 hours, collecting primary ethanol extract, soaking blue-green microalgae with 75 v% ethanol 5 times of the amount of the ethanol for 24 hours, collecting secondary ethanol extract, combining the two ethanol extracts, removing ethanol, freeze-drying residues to obtain freeze-dried substance, grinding, and sieving to obtain the final product.
The preparation method of the avocado fruit extract comprises the following steps: removing core and peel of avocado fruit, oven drying and pulverizing pulp, adding 10 times of 85 v% ethanol, ultrasonic processing for 3 hr, removing ethanol, freeze drying the residue to obtain lyophilized extract, grinding, and sieving.
Example 2
Example 2 provides a composition for targeted scar repair construction, comprising, by weight, 40 parts of an India swertia extract, 45 parts of a sweet orange fruit extract, 30 parts of a blue-green microalgae extract, and 22 parts of a avocado fruit extract.
The preparation method of the India swertia extract comprises the following steps:
a. callus culture: taking tender leaves of India swertia as explants to carry out plant tissue culture on an MS solid culture medium, and collecting the growing callus blocks;
b. seed cell culture: taking 5g of callus blocks, adding 125mL of 0.1g/L pectinase solution to treat the callus blocks, filtering, centrifugally separating, collecting seed cells, inoculating the seed cells to an MS liquid culture medium for culture, culturing at room temperature for 7 days, then carrying out amplification culture, carrying out amplification culture in a fermentation tank, culturing for 15 days, and centrifugally collecting the cultured cells;
c. preparing an extract: taking 50g of cultured cells, extracting for 2 times by using 70 v% ethanol, combining extracting solutions, filtering to obtain a crude extract, removing the ethanol to obtain an extract, mixing the extract with silica gel according to a weight ratio of 1: 4 mixing, loading into a chromatographic column, and mixing by using a mixture prepared by mixing the components in a volume ratio of 10: 1, eluting with an ethyl acetate/ethanol mixture, and then eluting with a solvent with a volume ratio of 8: 1, and finally eluting with an ethyl acetate/ethanol mixture in a volume ratio of 2: eluting with ethyl acetate/ethanol mixed solution of 1 to obtain purified eluent, and removing solvent to obtain the final product.
The preparation method of the sweet orange fruit extract comprises the following steps: cleaning sweet orange fruits, slicing, drying, pulverizing to obtain powder, adding 12 times of water, placing in an ultra-low temperature refrigerator, freezing at-70 deg.C, thawing for 3min under microwave heating at 70 deg.C, thawing the frozen solution for 1h under ultrasonic oscillation, centrifuging to obtain supernatant, filtering, concentrating, passing through a column with snake cage resin, washing the resin column with water, eluting with 70 v% ethanol to obtain eluate, removing solvent, freeze drying, and grinding.
The preparation method of the blue-green microalgae extract comprises the following steps: soaking blue-green microalgae with 85 v% ethanol 8 times of the amount of the ethanol for 24 hours, collecting primary ethanol extract, soaking blue-green microalgae with 75 v% ethanol 5 times of the amount of the ethanol for 24 hours, collecting secondary ethanol extract, combining the two ethanol extracts, removing ethanol, freeze-drying residues to obtain freeze-dried substance, grinding, and sieving to obtain the final product.
The preparation method of the avocado fruit extract comprises the following steps: removing core and peel of avocado fruit, oven drying and pulverizing pulp, adding 10 times of 85 v% ethanol, ultrasonic processing for 3 hr, removing ethanol, freeze drying the residue to obtain lyophilized extract, grinding, and sieving.
Example 3
Example 3 provides a composition for targeted scar repair construction, comprising 70 parts by weight of an India swertia extract, 25 parts by weight of a sweet orange fruit extract, 12 parts by weight of a blue-green microalgae extract, and 30 parts by weight of a avocado fruit extract.
The preparation method of the India swertia extract comprises the following steps:
a. callus culture: taking tender leaves of India swertia as explants to carry out plant tissue culture on an MS solid culture medium, and collecting the growing callus blocks;
b. seed cell culture: taking 5g of callus blocks, adding 125mL of 0.1g/L pectinase solution to treat the callus blocks, filtering, centrifugally separating, collecting seed cells, inoculating the seed cells to an MS liquid culture medium for culture, culturing at room temperature for 7 days, then carrying out amplification culture, carrying out amplification culture in a fermentation tank, culturing for 15 days, and centrifugally collecting the cultured cells;
c. preparing an extract: taking 50g of cultured cells, extracting for 2 times by using 70 v% ethanol, combining extracting solutions, filtering to obtain a crude extract, removing the ethanol to obtain an extract, mixing the extract with silica gel according to a weight ratio of 1: 4 mixing, loading into a chromatographic column, and mixing by using a mixture prepared by mixing the components in a volume ratio of 10: 1, eluting with an ethyl acetate/ethanol mixture, and then eluting with a solvent with a volume ratio of 8: 1, and finally eluting with an ethyl acetate/ethanol mixture in a volume ratio of 2: eluting with ethyl acetate/ethanol mixed solution of 1 to obtain purified eluent, and removing solvent to obtain the final product.
The preparation method of the sweet orange fruit extract comprises the following steps: cleaning sweet orange fruits, slicing, drying, pulverizing to obtain powder, adding 12 times of water, placing in an ultra-low temperature refrigerator, freezing at-70 deg.C, thawing for 3min under microwave heating at 70 deg.C, thawing the frozen solution for 1h under ultrasonic oscillation, centrifuging to obtain supernatant, filtering, concentrating, passing through a column with snake cage resin, washing the resin column with water, eluting with 70 v% ethanol to obtain eluate, removing solvent, freeze drying, and grinding.
The preparation method of the blue-green microalgae extract comprises the following steps: soaking blue-green microalgae with 85 v% ethanol 8 times of the amount of the ethanol for 24 hours, collecting primary ethanol extract, soaking blue-green microalgae with 75 v% ethanol 5 times of the amount of the ethanol for 24 hours, collecting secondary ethanol extract, combining the two ethanol extracts, removing ethanol, freeze-drying residues to obtain freeze-dried substance, grinding, and sieving to obtain the final product.
The preparation method of the avocado fruit extract comprises the following steps: removing core and peel of avocado fruit, oven drying and pulverizing pulp, adding 10 times of 85 v% ethanol, ultrasonic processing for 3 hr, removing ethanol, freeze drying the residue to obtain lyophilized extract, grinding, and sieving.
Example 4
Example 4 provides a composition for targeted scar repair construction, comprising 45 parts by weight of an India swertia extract, 30 parts by weight of a sweet orange fruit extract, 22 parts by weight of a blue-green microalgae extract, and 40 parts by weight of a avocado fruit extract.
The preparation method of the India swertia extract comprises the following steps:
a. callus culture: taking tender leaves of India swertia as explants to carry out plant tissue culture on an MS solid culture medium, and collecting the growing callus blocks;
b. seed cell culture: taking 5g of callus blocks, adding 125mL of 0.1g/L pectinase solution to treat the callus blocks, filtering, centrifugally separating, collecting seed cells, inoculating the seed cells to an MS liquid culture medium for culture, culturing at room temperature for 7 days, then carrying out amplification culture, carrying out amplification culture in a fermentation tank, culturing for 15 days, and centrifugally collecting the cultured cells;
c. preparing an extract: taking 50g of cultured cells, extracting for 2 times by using 70 v% ethanol, combining extracting solutions, filtering to obtain a crude extract, removing the ethanol to obtain an extract, mixing the extract with silica gel according to a weight ratio of 1: 4 mixing, loading into a chromatographic column, and mixing by using a mixture prepared by mixing the components in a volume ratio of 10: 1, eluting with an ethyl acetate/ethanol mixture, and then eluting with a solvent with a volume ratio of 8: 1, and finally eluting with an ethyl acetate/ethanol mixture in a volume ratio of 2: eluting with ethyl acetate/ethanol mixed solution of 1 to obtain purified eluent, and removing solvent to obtain the final product.
The preparation method of the sweet orange fruit extract comprises the following steps: cleaning sweet orange fruits, slicing, drying, pulverizing to obtain powder, adding 12 times of water, placing in an ultra-low temperature refrigerator, freezing at-70 deg.C, thawing for 3min under microwave heating at 70 deg.C, thawing the frozen solution for 1h under ultrasonic oscillation, centrifuging to obtain supernatant, filtering, concentrating, passing through a column with snake cage resin, washing the resin column with water, eluting with 70 v% ethanol to obtain eluate, removing solvent, freeze drying, and grinding.
The preparation method of the blue-green microalgae extract comprises the following steps: soaking blue-green microalgae with 85 v% ethanol 8 times of the amount of the ethanol for 24 hours, collecting primary ethanol extract, soaking blue-green microalgae with 75 v% ethanol 5 times of the amount of the ethanol for 24 hours, collecting secondary ethanol extract, combining the two ethanol extracts, removing ethanol, freeze-drying residues to obtain freeze-dried substance, grinding, and sieving to obtain the final product.
The preparation method of the avocado fruit extract comprises the following steps: removing core and peel of avocado fruit, oven drying and pulverizing pulp, adding 10 times of 85 v% ethanol, ultrasonic processing for 3 hr, removing ethanol, freeze drying the residue to obtain lyophilized extract, grinding, and sieving.
Example 5
Example 5 provides a composition for targeted scar repair construction, comprising 65 parts by weight of an India swertia extract, 40 parts by weight of a sweet orange fruit extract, 17 parts by weight of a blue-green microalgae extract, and 15 parts by weight of a avocado fruit extract.
The preparation method of the India swertia extract comprises the following steps:
a. callus culture: taking tender leaves of India swertia as explants to carry out plant tissue culture on an MS solid culture medium, and collecting the growing callus blocks;
b. seed cell culture: taking 5g of callus blocks, adding 125mL of 0.1g/L pectinase solution to treat the callus blocks, filtering, centrifugally separating, collecting seed cells, inoculating the seed cells to an MS liquid culture medium for culture, culturing at room temperature for 7 days, then carrying out amplification culture, carrying out amplification culture in a fermentation tank, culturing for 15 days, and centrifugally collecting the cultured cells;
c. preparing an extract: taking 50g of cultured cells, extracting for 2 times by using 70 v% ethanol, combining extracting solutions, filtering to obtain a crude extract, removing the ethanol to obtain an extract, mixing the extract with silica gel according to a weight ratio of 1: 4 mixing, loading into a chromatographic column, and mixing by using a mixture prepared by mixing the components in a volume ratio of 10: 1, eluting with an ethyl acetate/ethanol mixture, and then eluting with a solvent with a volume ratio of 8: 1, and finally eluting with an ethyl acetate/ethanol mixture in a volume ratio of 2: eluting with ethyl acetate/ethanol mixed solution of 1 to obtain purified eluent, and removing solvent to obtain the final product.
The preparation method of the sweet orange fruit extract comprises the following steps: cleaning sweet orange fruits, slicing, drying, pulverizing to obtain powder, adding 12 times of water, placing in an ultra-low temperature refrigerator, freezing at-70 deg.C, thawing for 3min under microwave heating at 70 deg.C, thawing the frozen solution for 1h under ultrasonic oscillation, centrifuging to obtain supernatant, filtering, concentrating, passing through a column with snake cage resin, washing the resin column with water, eluting with 70 v% ethanol to obtain eluate, removing solvent, freeze drying, and grinding.
The preparation method of the blue-green microalgae extract comprises the following steps: soaking blue-green microalgae with 85 v% ethanol 8 times of the amount of the ethanol for 24 hours, collecting primary ethanol extract, soaking blue-green microalgae with 75 v% ethanol 5 times of the amount of the ethanol for 24 hours, collecting secondary ethanol extract, combining the two ethanol extracts, removing ethanol, freeze-drying residues to obtain freeze-dried substance, grinding, and sieving to obtain the final product.
The preparation method of the avocado fruit extract comprises the following steps: removing core and peel of avocado fruit, oven drying and pulverizing pulp, adding 10 times of 85 v% ethanol, ultrasonic processing for 3 hr, removing ethanol, freeze drying the residue to obtain lyophilized extract, grinding, and sieving.
Evaluation of Performance
1. Cell proliferation efficacy test
1.1 cell inoculation: fibroblasts were seeded at 3.5E3 cells/well in 96-well plates, incubators (37 ℃, 5% CO)295% RH) overnight.
1.2, liquid preparation: sample working solutions with different concentrations were prepared according to the test concentration setting table (table 3).
TABLE 1 test concentration setting Table
Sample name | ① | ② | ③ | ④ | ⑤ | ⑥ | ⑦ | ⑧ |
Plant activating extract composition | 0.0078 | 0.0156 | 0.0313 | 0.0625 | 0.125 | 0.25 | 0.5 | 1 |
1.3 administration: according to the experimental grouping design of table 1, when the plating rate of the cells in the 96-well plate reaches 20% -30%, the drug is administered in groups, the dose of each hole is 200 μ L, and each group is provided with 3 multiple holes. Incubator (37 ℃, 5% CO)295% RH) for 24h, 48h and 72h respectively.
1.4, detection: after 24h of cell incubation and culture, discarding the supernatant, adding MTT working solution (0.5mg/mL, which is prepared for use now), incubating at 37 ℃ in dark place for 4h, discarding the supernatant after incubation, adding 150 mu LDMSO to each well, reading OD value at 490nm, and reading 96-well plates with 48h and 72h
The half-exchange treatment was carried out for 48 hours and 72 hours, and then the MTT assay was carried out.
1.5 cell relative viability calculation: calculating according to a formula:
2. barrier damage repair efficacy detection
2.1 design of the experiment
The experimental grouping was specifically set up as shown in table 2.
TABLE 2 Experimental design
2.2. Sample preparation
1) 0.4% SLS mother liquor configuration: 0.0080SLS is weighed and dissolved in 2ml PBS solution, and 0.22 mu m filtration is carried out to prepare 0.4 percent SLS mother liquor for standby.
2) 0.2% SLS working solution preparation: sucking 0.5ml of 0.4% SLS solution, and adding 0.5ml of PBS to prepare 0.2% SLS working solution.
3) Positive control group (WY14643) working solution formulation: weighing 10mg of WY14643 powder, dissolving in 1mLDMSO, and preparing into 30mMWY14643 mother liquor; to 5mL of the model culture solution, 8.3. mu.L of WY14643 mother liquor (30mM) was added to prepare a working solution of 50. mu.M for use.
4) Sample preparation: 25mg of sample is weighed and dissolved in 5mL of PBS to prepare a mother solution of 5mg/mL, and the mother solution is filtered for later use. And sucking 500 mu L of sample mother liquor, adding 500 mu L of 0.4% SLS mother liquor, and uniformly mixing for later use.
2.3 administration procedures
1) According to the experimental design of table 2, the model was transferred to a 6-well plate (0.9 mL of model culture solution was added in advance), and the 6-well plate was marked with a test group number.
2)Adding 25 μ L of prepared working solution to the surface of the model, adding 0.2% SLS solution to the surface of NC and PC groups, adding working solution with corresponding concentration to the sample group, clamping nylon membrane with tweezers to cover the surface of each model, uniformly distributing the samples on the surface of the model, placing in CO2Incubator (37 ℃, 5% CO)295% RH) for 24 h.
3) After the incubation is finished, the test object remained on the surface of the model is washed by a wash bottle filled with sterile PBS solution, and the residual liquid inside and outside the model is lightly wiped off by a sterile cotton swab.
2.4 inflammatory factor detection
After incubation for 24h, 3D skin model culture supernatant was collected in an EP tube, and after collection, samples for ELISA detection were stored in a freezer at-80 ℃. Detection analysis was performed according to the instructions of the ELISA kit.
3. Human clinical test
Mixing the composition obtained in the embodiments 1-5 with carbomer and water to prepare gel, wherein the weight ratio of the composition to the carbomer to the water is 20: 0.5: 10. clinical tests were conducted on 50 striae gravidarum testers with different degrees and different physical conditions, the striae gravidarum testers were divided into 5 groups, 5 groups of the striae gravidarum testers tried out gels prepared by using the compositions obtained in examples 1-5 respectively, the gel was coated once in the morning and at the evening of the striae gravidarum and persisted for 7 days, the effect of the compositions was significantly improved, slightly improved and not improved according to subjective evaluation of the testers, and the results are shown in table 3.
TABLE 3
Examples | Is remarkably improved | Slight improvement | Without improvement |
Example 1 | 9 | 1 | 0 |
Example 2 | 4 | 3 | 3 |
Example 3 | 5 | 2 | 3 |
Example 4 | 3 | 5 | 2 |
Example 5 | 5 | 1 | 4 |
According to the tests, the composition for constructing the target repair scar tissue network provided by the invention utilizes the scientific proportion of the plant active extract compositions such as the India swertia extract, the sweet orange fruit extract, the blue-green microalgae extract, the avocado fruit extract and the like, and various plant active substances cooperate with a target spot to repair, so that high-purity and high-activity effects are exerted on the skin, and the striae gravidarum repair of the accurate target spot repair is realized by applying a cell transmembrane technology; supplementing collagen, enhancing skin elasticity, and accelerating the recovery speed of the broken elastic fibers; four layers of skin have corresponding unique targeting mechanisms from subcutaneous tissue, dermis layer, epidermal layer and cuticle to repair target cells and tissues by aiming at a reaction mechanism.
The composition for constructing the target repair scar tissue mesh has the advantages that the action mechanism is clear, the trouble of striae gravidarum on married and fertile women can be well solved, the composition is based on solving the problem of striae gravidarum, cells are awakened, scar formation congestion is relieved, epithelial cells are activated, subcutaneous cells are acted, deep memory of skin is awakened, various damaged tissues are decomposed by penetrating into a dermis layer, the affected part is prevented from expanding outwards, efficient nutrition is provided to the cells through ultra-strong penetration, the damaged cytochromes are prevented from generating variation, and the purpose of eliminating the striae gravidarum is finally achieved; in addition, the traditional Chinese medicine composition can accelerate abdominal heat energy, consume fat, remove excessive water, promote metabolic function, improve the smoothness and the delicacy of the skin of the waist and the abdomen, increase the toughness of the skin, sculpt an abdominal curve, repair skin water locking, repair skin elastic fibers, promote collagen regeneration, and recover the elasticity, the flexibility and the compactness of the skin.
The foregoing examples are merely illustrative and serve to explain some of the features of the method of the present invention. The appended claims are intended to claim as broad a scope as is contemplated, and the examples presented herein are merely illustrative of selected implementations in accordance with all possible combinations of examples. Accordingly, it is applicants' intention that the appended claims are not to be limited by the choice of examples illustrating features of the invention. The use of some numerical ranges in the claims also includes sub-ranges within their range, and variations in these ranges are also to be construed as being covered by the appended claims where possible.
Claims (10)
1. A targeted scar repairing netting construction composition is characterized by comprising India swertia extract, sweet orange fruit extract, blue-green microalgae extract and avocado fruit extract.
2. The composition for targeted scar tissue mesh repair construction according to claim 1, wherein the weight ratio of the India swertia extract, the sweet orange fruit extract, the blue-green microalgae extract and the avocado fruit extract is (40-70): (25-45): (12-30): (15-40).
3. The composition for targeted scar tissue mesh repair construction according to claim 2, wherein the weight ratio of the India swertia extract, the sweet orange fruit extract, the blue-green microalgae extract and the avocado fruit extract is (45-65): (30-40): (15-25): (20-35).
4. The composition for targeted scar tissue mesh repair construction according to claim 3, wherein the weight ratio of the India swertia extract, the sweet orange fruit extract, the blue-green microalgae extract and the avocado fruit extract is (50-60): (32-38): (18-22): (22-32).
5. The composition for the targeted scar tissue net construction repair according to any one of claims 1 to 4, characterized in that the method for preparing the India swertia extract comprises the following steps:
a. callus culture: taking tender leaves of India swertia as explants to carry out plant tissue culture, and collecting the growing callus blocks;
b. seed cell culture: separating the callus blocks by an enzymolysis method, filtering, separating, collecting seed cells, and inoculating the seed cells to a culture medium for culture;
c. preparing an extract: adding ethanol into cultured cells, extracting to obtain crude extract, removing solvent, and purifying with chromatographic column.
6. The targeted scar tissue mesh construction repair composition of any one of claims 1 to 4 wherein the preparation method of the sweet orange fruit extract comprises the following steps: cleaning fructus Citri sinensis, slicing, oven drying, pulverizing to obtain powder, adding water, freezing, thawing under microwave heating, ultrasonic vibrating, centrifuging to obtain supernatant, filtering, concentrating, eluting with snake cage resin, removing solvent, freeze drying, and grinding.
7. The composition for targeted scar tissue mesh repair construction according to any one of claims 1 to 4, wherein the preparation method of the blue-green microalgae extract comprises the following steps: soaking the blue-green microalgae with 80-95 v% ethanol for 24 hours, collecting a primary ethanol extract, soaking the blue-green microalgae with 70-80 v% ethanol for 24 hours, collecting a secondary ethanol extract, combining the two ethanol extracts, removing ethanol, freeze-drying the remainder to obtain a freeze-dried substance, grinding and sieving to obtain the product.
8. The composition for targeted scar tissue mesh repair construction according to any one of claims 1 to 4, wherein the avocado fruit extract is prepared by a method comprising the following steps: removing kernels and peels of avocado fruits, drying and crushing the avocado fruits, adding a solvent, carrying out ultrasonic treatment for 3-4 hours, removing the solvent, freeze-drying, and grinding to obtain the avocado fruit extract.
9. Use of the composition for targeted scar tissue mesh construction according to any one of claims 1 to 8 for preventing or repairing stretch marks.
10. The use of the composition for targeted scar tissue mesh construction repair according to claim 9 wherein the composition is used in the preparation of gels, creams, lotions, essential oils.
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