CN110652491A - Skin care product with effect of removing striae gravidarum and preparation method and application thereof - Google Patents
Skin care product with effect of removing striae gravidarum and preparation method and application thereof Download PDFInfo
- Publication number
- CN110652491A CN110652491A CN201910938897.5A CN201910938897A CN110652491A CN 110652491 A CN110652491 A CN 110652491A CN 201910938897 A CN201910938897 A CN 201910938897A CN 110652491 A CN110652491 A CN 110652491A
- Authority
- CN
- China
- Prior art keywords
- parts
- stem cell
- skin care
- mesenchymal stem
- care product
- 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.)
- Granted
Links
Images
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
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/67—Vitamins
- A61K8/673—Vitamin B group
- A61K8/675—Vitamin B3 or vitamin B3 active, e.g. nicotinamide, nicotinic acid, nicotinyl aldehyde
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/67—Vitamins
- A61K8/676—Ascorbic acid, i.e. vitamin C
-
- 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
- A61K8/72—Cosmetics or similar toiletry preparations characterised by the composition containing organic macromolecular compounds
- A61K8/73—Polysaccharides
- A61K8/731—Cellulose; Quaternized cellulose derivatives
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/92—Oils, fats or waxes; Derivatives thereof, e.g. hydrogenation products thereof
- A61K8/922—Oils, fats or waxes; Derivatives thereof, e.g. hydrogenation products thereof of vegetable origin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
- A61K8/9783—Angiosperms [Magnoliophyta]
- A61K8/9789—Magnoliopsida [dicotyledons]
-
- 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
-
- 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
- A61Q19/02—Preparations for care of the skin for chemically bleaching or whitening the skin
-
- 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
-
- 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
- A61K2800/70—Biological properties of the composition as a whole
- A61K2800/72—Hypo-allergenic
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- General Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Epidemiology (AREA)
- Birds (AREA)
- Dermatology (AREA)
- Biotechnology (AREA)
- Engineering & Computer Science (AREA)
- Mycology (AREA)
- Microbiology (AREA)
- Botany (AREA)
- Tropical Medicine & Parasitology (AREA)
- Chemical & Material Sciences (AREA)
- Oil, Petroleum & Natural Gas (AREA)
- Gerontology & Geriatric Medicine (AREA)
- Cosmetics (AREA)
Abstract
The invention relates to a skin care product with a stretch mark removing effect and a preparation method and application thereof, belonging to the technical field of cosmetics. The skin care product provided by the invention comprises the following raw materials in parts by volume: 10-25 parts of a basjoo flower extracting solution, 20-45 parts of a stem cell extract, 5-12 parts of camellia oil, 1-5 parts of nicotinamide, 1-5 parts of vitamin C, 0.5-3 parts of panthenol, 1-4 parts of PEG-40 hydrogenated castor oil, 0.5-2 parts of a preservative, 0.1-0.8 part of hydroxyethyl cellulose and 5-60 parts of deionized water. The skin care product provided by the invention has the advantages that the stem cell exosomes act on subcutaneous fibroblasts, the fibroblast proliferation and repair and the peripheral blood vessel microenvironment improvement are emphasized, and the skin care product has good skin conditioning effects of anti-inflammation, moisture preservation, whitening and the like by adding natural plant components such as the Barbadia extract and vitamin nutrient substances, so that the striae gravidarum improvement effect is achieved.
Description
Technical Field
The invention relates to a skin care product with a stretch mark removing effect and a preparation method and application thereof, belonging to the technical field of cosmetics.
Background
The abdominal skin of women is gradually stretched during pregnancy, resulting in destructive damage to the connective tissue of the dermis (mainly collagen fibers and elastic fibers), weakening the extensibility and elasticity of the skin, and then causing striated skin lesions after delivery. Clinically, it is initially manifested as pale red or purple stripes, gradually turning into pale white with striae atrophicae and fine lines. In reality, stretch marks represent more of a common cosmetic problem of skin in women. Since the striations, once formed, are not effectively completely eliminated, but are improved as much as possible. At present, the solution is basically to apply olive oil, skin lotion and other preventive means during pregnancy, or to locally use all-trans vitamin A acid or tartaric acid compounds, or to eliminate and fade striae gravidarum by laser means, but the effect is not good. Based on the pathophysiology of striae gravidarum, the prevention and reduction of striae gravidarum related to pregnancy requires the use of targeting skin fibroblasts, and the problem of striae gravidarum can be solved only effectively by activating the proliferation of skin fibroblasts and stimulating the metabolism, in particular increasing the synthesis of subcutaneous elastic fibers and collagen structures.
Disclosure of Invention
The invention aims to overcome the defects of the prior art and provide a skin care product which is mild, moist, comfortable, non-irritant and capable of effectively fading stretch marks.
It is another object of the present invention to provide a method of making the skin care article.
The invention further aims to provide the application of the skin care product in preparing a striae gravidarum removing product.
In order to achieve the purpose, the invention adopts the technical scheme that: a skin care product with a stretch mark removing effect comprises the following raw materials in parts by volume: 10-25 parts of a basjoo flower extracting solution, 20-45 parts of a stem cell extract, 5-12 parts of camellia oil, 1-5 parts of nicotinamide, 1-5 parts of vitamin C, 0.5-3 parts of panthenol, 1-4 parts of PEG-40 hydrogenated castor oil, 0.5-2 parts of a preservative, 0.1-0.8 part of hydroxyethyl cellulose and 5-60 parts of deionized water.
Preferably, the skin care product comprises the following raw materials in parts by volume: 20 parts of basjoo flower extract, 40 parts of stem cell extract, 10 parts of camellia oil, 5 parts of nicotinamide, 5 parts of vitamin C, 2 parts of panthenol, 3 parts of PEG-40 hydrogenated castor oil, 2 parts of preservative, 0.7 part of hydroxyethyl cellulose and 15 parts of deionized water.
Preferably, the skin care article further comprises citric acid or sodium citrate.
Preferably, the stem cell extract comprises disrupted mesenchymal stem cell exosomes and a stem cell culture supernatant.
In addition, the invention provides a preparation method of the skin care product, which comprises the following steps:
(1) weighing the raw materials according to the formula;
(2) putting nicotinamide and vitamin C into a container, adding deionized water to fully dissolve, adding the basjoa flower extract, and stirring and mixing uniformly;
(3) adding hydroxyethyl cellulose into the solution obtained in the step (2), fully dissolving and stirring until the mixture is homogeneous;
(4) adding camellia oil, panthenol and PEG-40 hydrogenated castor oil into the solution obtained in the step (3), and stirring to homogenize;
(5) adding the stem cell extract into the solution obtained in the step (4), stirring until the stem cell extract is uniformly mixed, then adding the preservative, and stirring and uniformly mixing;
(6) and (5) adding citric acid or sodium citrate into the solution obtained in the step (5) to adjust the pH value, thus obtaining the skin care product.
In a preferred embodiment of the preparation method of the skin care product, the preparation method of the basjoo flower extract comprises the following steps: extracting fresh flos Bombacis Cannabifolii under reflux for 3 times; extracting flos Bombacis Cannabinae and 65% ethanol at a mass ratio of 1:18 for 1.5h at the 1 st time; extracting flos Bombacis Cannabinae and 60% ethanol at a mass ratio of 1:10 for 1.5h for the second time; extracting flos Bombacis Cannabinae and 60% ethanol at a mass ratio of 1:8 for 1.5h at the 3 rd time; filtering the extractive solutions extracted for 3 times with three layers of gauze, mixing filtrates, and concentrating the filtrate under reduced pressure to 5 times of the weight of flos Bombacis Cannabinae to obtain the final product.
In a preferred embodiment of the method for preparing the skin care product of the present invention, the preservative is prepared by: weighing 6 parts of methyl p-hydroxybenzoate, adding a 17-time volume of 1, 3-butanediol solution, stirring until the solution is clear, and fully dissolving to obtain the preservative.
In a preferred embodiment of the method for preparing a skin care product according to the present invention, the pH in step (6) is 6.
In a preferred embodiment of the method for preparing a skin care preparation of the present invention, the method for preparing the stem cell extract comprises the following steps:
(1) carrying out in-vitro large-scale culture on stem cells: culturing according to GMP cell to obtain mesenchymal stem cell;
(2) carrying out ultrasonic crushing on the mesenchymal stem cells collected after the culture in the step (1) to obtain a crushed mesenchymal stem cell exosome;
(3) collecting the culture supernatant of the mesenchymal stem cells in the step (1) to obtain a stem cell culture supernatant exosome;
(4) and (3) respectively dissolving the crushed mesenchymal stem cell exosomes in the step (2) and the stem cell culture supernatant exosomes in the step (3) in normal saline, mixing in equal volume, and stirring uniformly to obtain the stem cell extract.
In a preferred embodiment of the method for preparing a skin care product according to the present invention, in the step (1), the mesenchymal stem cells are expanded by using a cell factory (ten layers of GMP grade) equipment according to a GMP cell culture process, and 3 × 10 cells can be harvested when the cell fusion rate reaches 95% or more8The mesenchymal stem cell of (a); in the step (2), the mesenchymal stem cells are subjected to ultrasonic disruption in a specific operation of 1 × 108Adding the cell number into 100ml of physiological saline, centrifuging at 4 ℃ for 10min at 300 Xg to remove cell debris, and collecting the supernatant; centrifuging at 4 deg.C 2500 Xg for 30min to remove apoptotic bodies, and collecting supernatant; centrifuging at 13000 Xg for 1 hr to remove platelet impurities, and collectingCollecting the supernatant; centrifuging at 4 deg.C 100000 × g for 70min to obtain the crushed exosome of mesenchymal stem cell; in the step (3), the specific operation of collecting the mesenchymal stem cell culture supernatant is as follows: centrifuging the mesenchymal stem cells in the step (1) at the temperature of 4 ℃ of 2000 Xg for 10min to remove cell debris, and collecting supernatant; centrifuging at 13000 Xg for 1h to remove platelet impurities, and collecting the supernatant; centrifuging at 4 deg.C 100000 × g for 70min to obtain exosome of stem cell culture supernatant; in the step (4), the protein concentration of the stem cell extract is 40 mg/ml.
In a preferred embodiment of the preparation method of the skin care product, the protein concentration of the disrupted mesenchymal stem cell exosomes of step (2) and the protein concentration of the stem cell culture supernatant exosomes of step (3) are respectively measured by BCA protein concentration measurement, wherein the protein concentration measured in step (2) is 9.5mg/ml, and the protein concentration measured in step (3) is 53.6 mg/ml.
In addition, the invention provides the application of the skin care product in preparing a striae gravidarum removing product.
Compared with the prior art, the invention has the beneficial effects that:
(1) the skin care product with the effect of removing the stretch marks, provided by the invention, has the advantages that the exosome protein in the component stem cell extract can effectively promote the proliferation of fibroblasts, synthesize and secrete collagen fibers, elastic fibers and reticular fibers, increase the content of the elastic fibers and the collagen structure in the skin, and effectively prevent the stretch marks or improve the symptoms of the stretch marks;
(2) the skincare product with the effect of removing striae gravidarum is added with the basjo flower extracting solution and camellia oil natural plant essence, the basjo flower extracting solution contains various aromatic alcohol compounds such as aromatic alcohol and geraniol, and has the effects of resisting inflammation, relieving itching, replenishing water and repairing, and camellia oil contains rich natural vitamin E, tea polyphenol, unsaturated fatty acid, squalene and the like and has the effects of beautifying and beautifying skin;
(3) the skin care product with the effect of removing stretch marks, provided by the invention, also contains vitamin C necessary for collagen synthesis, promotes collagen synthesis, and also contains vitamin B components such as nicotinamide and panthenol, wherein the nicotinamide whitens and improves skin texture, and the panthenol is a precursor of pantothenic acid (vitamin B5), can improve skin barrier, promotes fibroblast proliferation and re-epithelization of epidermis, and is also a small-molecule natural moisturizing component;
(4) the skin care product with the effect of removing the striae gravidarum provided by the invention has the advantages that the contained stem cell exosomes act on subcutaneous fibroblasts, the proliferation and repair of the fibroblasts and the improvement of the microenvironment of peripheral blood vessels are emphasized, and the skin conditioning effects of good inflammation resistance, moisture retention, whitening and the like are realized by adding natural plant components such as the flower extract and vitamin nutrient substances, so that the effect of improving the striae gravidarum is achieved;
(5) the skin care product with the effect of removing the striae gravidarum provided by the invention adopts food-grade raw materials, does not contain antibiotics and hormones, and does not generate toxic or side effect.
Drawings
FIG. 1 is a graph showing the results of the change in striae gravidarum in the same part of a subject before and after 12 weeks using the skin care preparation of example 1 of the present invention;
FIG. 2 is a graph showing the change in striae gravidarum between subjects before and after 12 weeks using the skin care preparation of example 1 of the present invention on the same site.
Detailed Description
To better illustrate the objects, aspects and advantages of the present invention, the present invention will be further described with reference to specific examples.
Example 1
The embodiment is a skin care product with the effect of removing stretch marks, which comprises the following raw materials in parts by volume: 20 parts of basjoo flower extract, 40 parts of stem cell extract, 10 parts of camellia oil, 5 parts of nicotinamide, 5 parts of vitamin C, 2 parts of panthenol, 3 parts of PEG-40 hydrogenated castor oil, 2 parts of preservative, 0.7 part of hydroxyethyl cellulose, 12 parts of deionized water and 0.3 part of sodium citrate.
The preparation method of the skin care product comprises the following steps:
(1) weighing the raw materials according to the formula;
(2) putting 5 parts of nicotinamide and 5 parts of vitamin C into a container, adding 12 parts of deionized water for full dissolution, adding 20 parts of the extract of the ballflower, and stirring and mixing uniformly;
(3) adding 0.7 part of hydroxyethyl cellulose into the solution in the step (2), fully dissolving and stirring to be homogeneous;
(4) adding 10 parts of camellia oil, 2 parts of panthenol and 3 parts of PEG-40 hydrogenated castor oil into the solution obtained in the step (3), and stirring until the mixture is homogeneous;
(5) adding 40 parts of the stem cell extract into the solution obtained in the step (4), stirring until the mixture is uniformly mixed, then adding 2 parts of the preservative, and stirring and uniformly mixing;
(6) adding 0.3 part of sodium citrate into the solution obtained in the step (5) to adjust the pH value to 6, thus obtaining the skin care product;
the preparation method of the basjoo flower extracting solution comprises the following steps: the method is operated by using a common reflux extraction method of traditional Chinese medicinal materials, namely extracting fresh flowers of the Chinese ballflower by refluxing for 3 times: extracting flos Bombacis Cannabinae and 65% ethanol at a mass ratio of 1:18 for 1.5h at the 1 st time; extracting flos Bombacis Cannabinae and 60% ethanol at a mass ratio of 1:10 for 1.5h for the second time; extracting flos Bombacis Cannabinae and 60% ethanol at a mass ratio of 1:8 for 1.5h at the 3 rd time; filtering the extractive solutions extracted for 3 times with three layers of gauze, mixing filtrates, and concentrating the filtrate under reduced pressure to 5 times of the weight of flos Bombacis chinensis;
the preparation method of the stem cell extract comprises the following steps:
(1) carrying out in-vitro large-scale culture on stem cells: according to GMP conventional cell culture operation flow, using cell factory (ten layers of GMP grade) equipment to expand mesenchymal stem cells, and obtaining 3 × 10 cells when cell fusion rate reaches above 95%8The mesenchymal stem cell of (a);
(2) carrying out ultrasonic disruption on the mesenchymal stem cells collected after the culture in the step (1): by 1 × 108Adding cell amount into 100ml physiological saline (0.9% sodium chloride aqueous solution), setting cell disruptor parameters to (600W; 4 deg.C; 2s intermittent 2s ultrasonic 10min), centrifuging at 300 Xg for 10min, removing cell debris at 4 deg.C, and collecting supernatant; centrifuging at 4 deg.C 2500 Xg for 30min to remove apoptotic bodies, and collecting supernatant; centrifuging at 13000 Xg for 1 hr to remove platelet impurities, collecting supernatant, and centrifuging at 100000 Xg at 4 deg.CObtaining an exosome precipitate after 70min, and carrying out BCA protein concentration determination on the obtained crushed mesenchymal stem cell exosome, wherein the concentration of the exosome precipitate is 9.5 mg/ml;
(3) collecting the culture supernatant of the mesenchymal stem cells in the step (1), centrifuging at 2000 Xg for 10min at 4 ℃ to remove cell debris, and collecting the supernatant; centrifuging at 13000 Xg for 1h to remove platelet impurities, and collecting the supernatant; centrifuging at 100000 Xg for 70min at 4 deg.C to obtain exosome precipitate, and measuring BCA protein concentration of the exosome in the cell culture supernatant to 53.6 mg/ml;
(4) and (3) respectively dissolving the crushed mesenchymal stem cell exosomes in the step (2) and the stem cell culture supernatant exosomes in the step (3) in normal saline, mixing in equal volume, and uniformly stirring to obtain a stem cell extract (the protein concentration is 40 mg/ml).
The preparation method of the preservative comprises the following steps: weighing 6 parts of methyl p-hydroxybenzoate, adding a 17-time volume of 1, 3-butanediol solution, stirring until the solution is clear, and fully dissolving to obtain the preservative.
Example 2
The embodiment is a skin care product with the effect of removing stretch marks, which comprises the following raw materials in parts by volume: 10 parts of basjoo flower extract, 20 parts of stem cell extract, 5 parts of camellia oil, 1 part of nicotinamide, 1 part of vitamin C, 0.5 part of panthenol, 1 part of PEG-40 hydrogenated castor oil, 0.5 part of preservative, 0.1 part of hydroxyethyl cellulose, 60 parts of deionized water and 0.9 part of sodium citrate.
The preparation method of the skin care product comprises the following steps:
(1) weighing the raw materials according to the formula;
(2) putting 1 part of nicotinamide and 1 part of vitamin C into a container, adding 60 parts of deionized water for full dissolution, adding 10 parts of basjoo flower extract, and stirring and mixing uniformly;
(3) adding 0.1 part of hydroxyethyl cellulose into the solution obtained in the step (2), fully dissolving and stirring until the mixture is homogeneous;
(4) adding 5 parts of camellia oil, 0.5 part of panthenol and 1 part of PEG-40 hydrogenated castor oil into the solution obtained in the step (3), and stirring until the mixture is homogeneous;
(5) adding 20 parts of the stem cell extract into the solution obtained in the step (4), stirring until the mixture is uniformly mixed, then adding 0.5 part of preservative, and stirring and uniformly mixing;
(6) and (5) adding 0.9 part of sodium citrate into the solution obtained in the step (5) to adjust the pH value to 6, thus obtaining the skin care product.
Wherein the preparation method of the above basjoo flower extract, the preparation method of the above stem cell extract and the preparation method of the above preservative are the same as in example 1.
Example 3
The embodiment is a skin care product with the effect of removing stretch marks, which comprises the following raw materials in parts by volume: 25 parts of basjoo flower extract, 45 parts of stem cell extract, 12 parts of camellia oil, 2 parts of nicotinamide, 2 parts of vitamin C, 3 parts of panthenol, 4 parts of PEG-40 hydrogenated castor oil, 2 parts of preservative, 0.8 part of hydroxyethyl cellulose, 2.2 parts of deionized water and 0.3 part of citric acid.
The preparation method of the skin care product comprises the following steps:
(1) weighing the raw materials according to the formula;
(2) putting 5 parts of nicotinamide and 2 parts of vitamin C into a container, adding 2.2 parts of deionized water for full dissolution, adding 25 parts of Bombay flower extract, stirring and mixing uniformly
(3) Adding 0.8 part of hydroxyethyl cellulose into the solution obtained in the step (2), fully dissolving and stirring until the mixture is homogeneous;
(4) adding 12 parts of camellia oil, 3 parts of panthenol and 4 parts of PEG-40 hydrogenated castor oil into the solution obtained in the step (3), and stirring until the mixture is homogeneous;
(5) adding 45 parts of the stem cell extract into the solution obtained in the step (4), stirring until the mixture is uniformly mixed, then adding 2 parts of the preservative, and stirring and uniformly mixing;
(6) adding 0.3 part of citric acid into the solution obtained in the step (5) to adjust the pH value to 6, thus obtaining the skin care product;
wherein the preparation method of the above basjoo flower extract, the preparation method of the above stem cell extract and the preparation method of the above preservative are the same as in example 1.
Comparative example 1
The comparative example is a skin care product, which comprises the following raw materials in parts by volume: 10 parts of camellia oil, 5 parts of nicotinamide, 5 parts of vitamin C, 2 parts of panthenol, 3 parts of PEG-40 hydrogenated castor oil, 2 parts of preservative, 0.7 part of hydroxyethyl cellulose, 72 parts of deionized water and 0.3 part of sodium citrate.
(1) Weighing the raw materials according to the formula;
(2) putting 5 parts of nicotinamide and 5 parts of vitamin C into a container, adding 72 parts of deionized water to fully dissolve, and stirring and mixing uniformly;
(3) adding 0.7 part of hydroxyethyl cellulose into the solution in the step (2), fully dissolving and stirring to be homogeneous;
(4) adding 10 parts of camellia oil, 2 parts of panthenol and 3 parts of PEG-40 hydrogenated castor oil into the solution obtained in the step (3), and stirring until the mixture is homogeneous;
(5) adding 2 parts of preservative into the step (4), and stirring and mixing uniformly;
(6) adding 0.3 part of sodium citrate into the solution obtained in the step (5) to adjust the pH value to 6,
the preparation method of the preservative comprises the following steps: accurately weighing 6 parts of methyl p-hydroxybenzoate, adding a 17-time volume of 1, 3-butanediol solution, stirring until the solution is clear, and fully dissolving the solution.
Experimental example 1
Experimental example 1 shows the physical and chemical indexes of the skin care product of the invention example 1
And (3) quality detection: extruding the solution in the skin care product by adopting a random sampling method, and detecting according to a general cosmetic detection method:
(1) the skin care preparation of example 1 was subjected to pH measurement, and pH was measured to be 6-7.
(2) The skin care preparation of example 1 was refrigerated in a refrigerator at 4 ℃ and left in a thermostat at 37 ℃ for about one week, and was observed to have no delamination. Standing for 3 months, no layering phenomenon is observed, no change is caused after the use, no change is caused in smell, and no deterioration phenomenon is caused.
(3) The skin care article of example 1 was tested for microorganisms: and (4) refrigerating at room temperature and 4 ℃ for one week, 3 months and half a year respectively, and detecting no microorganism in the detection result.
Experimental example 2
The experimental example is a test of the striae gravidarum removing effect of the skin care product of example 1 and the skin care product of comparative example 1
Experimental data: 32 healthy female subjects treated with the skin care product prepared in example 1 and comparative example 1 of the present invention were studied, aged 26-40 years, and averaged (34.6 ± 3.0) years, from 2018 for 1 month to 2018 for 12 months. Inclusion criteria were: firstly, more than 3 months after delivery, striae gravidarum appears for 1-3 years, and striae gravidarum is obviously seen around the abdomen; the patient agrees to the study and signs an informed consent; and thirdly, no people who have not received other stretch mark treatment and are willing to receive follow-up visitors in the last year. Exclusion criteria: none of the following were included in the study: firstly, allergy to cosmetics, skin care products, topical medicaments and components thereof is known; ② women in lactation; the abdominal part has the skin abnormality such as inflammatory eczema, and systemic diseases can influence the research result;
the experimental method comprises the following steps: 32 subjects were selected, the abdomen was scrubbed with warm water before use, the peritoneum (skin care article of example 1 of the present invention) was attached to the subject site, the membrane paper was removed after 20 minutes, and the skin was lightly massaged for 10 minutes, 1 time per day for each subject. The evaluation was carried out 5 times before and after the test at 2 nd, 4 th, 8 th and 12 th weeks, respectively. The subjects divide the abdomen into two areas with the midline of the abdomen as a boundary, and select one side as the application part of the product according to the random principle, and use the skin care product of the comparative example 1 as the control group on the other side, and the application method is the same as that of the skin care product of the example 1. And selecting a stretch mark with similar severity degree as a marker on each side, and performing clinical judgment and instrument detection on the stretch marks.
Subjective evaluation indexes of the experiment are as follows: scores were given for striae gravidarum color, depth, and roughness, respectively, according to the SCAR scale, with the scoring criteria shown in Table 1.
TABLE 1 score criteria for striae gravidarum efficacy
Objective evaluation index of experiment:
measuring the area of the stretch mark: a commercially available preservative film is attached to the abdomen, a marker pen is used for tracing the striae gravidarum, then a scanner is used for scanning the film into pictures, and the traced striae gravidarum area is calculated by using Photoshop. Photoshop calculated traced target lesion area: and (3) correcting white balance by using a gray card in software, identifying and selecting an image of the striae gravidarum part, and measuring the area of the striae gravidarum by taking scales in a steel ruler as a scale.
Relevant parameters of the gestational skin texture: test procedure the skin total elasticity index (R2), the skin plastic recovery index (R5) values were measured at each time point using a skin elasticity tester. The images taken by the skin scanner show microscopic changes in the skin surface, the images are analyzed, and changes in the skin texture variation parameter (VAR), surface parameter (surface), and skin roughness parameter (C-R1) are measured.
Analysis of total effective rate of experiment: all data were analyzed with statistical software. The striae gravidarum area and the score of each item are expressed by the mean + -standard deviation. The self comparison before and after treatment adopts paired t test; the measurement data of the experimental side and the comparison side are tested by using independent samples t; p <0.05 indicates that the experimental result has significance statistics.
The experimental results are as follows: after the testee uses the product for 12 weeks, the clinical efficacy scores of the color, roughness and depth of the striae gravidarum before and after treatment have statistical significance (P is less than 0.05); correspondingly, the comparison of the parameters such as skin elasticity, surface texture and the like before and after treatment shows that the difference also has statistical significance (P is less than 0.05), the total effective rate of a test group is obviously higher than that of a control group, and the specific result (P is less than 0.05) is shown in tables 2-10.
TABLE 2 striae gravidarum Overall color score comparison
| Test group | Control group | |
| Base value | 3.19±0.90 | 3.31±0.93 |
| 2 weeks | 3.16±0.92 | 3.25±0.92 |
| 4 weeks | 3.03±0.92 | 3.13±0.83 |
| 8 weeks | 2.85±1.00 | 3.09±0.93 |
| For 12 weeks | 2.01±1.16* | 2.91±0.93 |
The overall severity of the striae gravidarum is scored through clinical judgment, and the result shows that after 12 weeks of treatment, the striae gravidarum condition of the test side part is significantly different from the basic value measured before the test and the striae gravidarum condition of the control side part, and after 12 weeks of treatment, the overall color score of the striae gravidarum of the test group is 2.01 +/-1.16 and is significantly lower than that of the control group, namely 2.91 +/-0.93, so that the skin care product of the example 1 containing the extract of the basjoram flower and the dry cell extract can effectively lighten the overall color of the striae gravidarum, and the lightening effect is more significant compared with the skin care product of the comparative example 1 containing no extract of the basjoram flower and the dry cell extract.
TABLE 3 striae gravidarum depth score comparison
| Test group | Control group | |
| Base line | 2.97±0.93 | 3.13±1.13 |
| 2 weeks | 2.88±0.86 | 3.09±1.06 |
| 4 weeks | 2.81±0.90 | 2.97±1.03 |
| 8 weeks | 2.73±0.98 | 3.00±1.08 |
| For 12 weeks | 2.52±1.08* | 2.98±1.18 |
TABLE 4 striae gravidarum area comparison
As can be seen from the results in tables 3 and 4, compared with the control group, the striae gravidarum area on the side of the test is gradually decreased by selecting the marked striae gravidarum area, the measured value after 8 weeks has significant difference from the basic value, and the control side has no significant difference at each time point, which indicates that the striae gravidarum on the skin is reduced and shrunk after the product is used. Thus, the skin care product of example 1 containing the extract solution of basjoo flower and the stem cell extract of the present invention can effectively lighten the depth of the striae gravidarum, reduce the area of the striae gravidarum, and has more remarkable lightening and shrinking effects compared with the skin care product of comparative example 1 containing no extract solution of basjoo flower and the stem cell extract.
TABLE 5 comparison of values of Total skin elasticity index (R2)
| Test group | Control group | |
| Radix | 0.80±0.10 | 0.79±0.08 |
| 2 weeks | 0.81±0.10 | 0.83±0.08 |
| 4 weeks | 0.85±0.10 | 0.83±0.09 |
| 8 weeks | 0.88±0.08* | 0.82±0.09 |
| For 12 weeks | 0.94±0.08* | 0.82±0.08 |
TABLE 6 comparison of values of skin plasticity recovery index (R5)
| Test group | Control group | |
| Base value | 0.83±0.15 | 0.84±0.13 |
| 2 weeks | 0.86±0.16 | 0.84±0.15 |
| 4 weeks | 0.88±0.16 | 0.83±0.13 |
| 8 weeks | 0.90±0.13* | 0.84±0.16 |
| For 12 weeks | 0.95±0.15* | 0.83±0.15 |
TABLE 7 comparison of skin texture variation values
TABLE 8 comparison of skin surface parameter variation values
| surface | Test group | Control group |
| Base line value | 7.21±1.54 | 7.12±1.83 |
| 2 weeks | 6.93±1.68 | 7.08±1.51 |
| 4 weeks | 6.41±1.48 | 7.10±1.76 |
| 8 weeks | 6.20±1.53* | 7.06±1.55 |
| For 12 weeks | 5.83±1.91* | 7.02±1.41 |
TABLE 9 comparison of skin roughness parameter variation values
| Test group | Control group | |
| Base line value | 73.50±4.37 | 74.00±4.18 |
| 2 weeks | 71.20±3.68 | 73.07±4.06 |
| 4 weeks | 70.10±3.46 | 74.06±4.02 |
| 8 weeks | 64.04±3.66* | 70.01±3.95 |
| For 12 weeks | 58.00±3.54* | 70.80±3.11 |
From the results of tables 5 to 9, it can be seen that the skin scanner takes images showing microscopic changes in the surface of the skin. The analysis finds that the baseline value of the control part and the baseline value of the test part have no obvious difference, and each parameter of the skin detector has obvious difference with the same group of basic value and the control part from the 8 th week. Each test time point data shows that the values of the total elasticity index (R2) and the skin plasticity recovery index (R5) of the skin elasticity detection of the test part are in rising trend; VAR, surface parameter value, C-R1 all decreased.
Wherein: VAR is a texture parameter, representing the local mean difference, the variation increases with wrinkle increase, surface is a surface parameter, reflecting the smoothness, the more wrinkles, the larger the surface value. C-R1 is a roughness parameter, reflecting a peripheral depth index, with values increasing the deeper the skin wrinkles.
As can be seen from fig. 1 and 2, two subjects were selected to observe the striae gravidarum lightening effect before and 12 weeks after the skin care product of example 1 of the present invention, and both fig. 1 and 2 show that the striae gravidarum removing effect is significant after 12 weeks of the skin care product of example 1 of the present invention.
TABLE 10 comparison of Total effective rates of striae gravidarum
| Invalidation | Is effective | Show effect | Basic cure | The total effective rate% | |
| Test group | 5 | 15 | 9 | 3 | 84.37 |
| Control group | 26 | 6 | 0 | 0 | 18.75 |
As can be seen from table 10 above, the skin care product of example 1 containing the extract solution of basca flower and the stem cell extract of the present invention has a total effective rate of reducing striae gravidarum of 84.37%, and the skin care product of comparative example 1 containing no extract solution of basca flower and the stem cell extract has a total effective rate of reducing striae gravidarum of 18.75%. Therefore, the striae gravidarum removing effect provided by the invention is remarkable.
Finally, it should be noted that the above embodiments are only used for illustrating the technical solutions of the present invention and not for limiting the protection scope of the present invention, and although the present invention is described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that modifications or equivalent substitutions can be made on the technical solutions of the present invention without departing from the spirit and scope of the technical solutions of the present invention.
Claims (10)
1. The skin care product with the effect of removing stretch marks is characterized by comprising the following raw materials in parts by volume: 10-25 parts of a basjoo flower extracting solution, 20-45 parts of a stem cell extract, 5-12 parts of camellia oil, 1-5 parts of nicotinamide, 1-5 parts of vitamin C, 0.5-3 parts of panthenol, 1-4 parts of PEG-40 hydrogenated castor oil, 0.5-2 parts of a preservative, 0.1-0.8 part of hydroxyethyl cellulose and 5-60 parts of deionized water.
2. The skin care article of claim 1, wherein the skin care article comprises the following parts by volume of raw materials: 20 parts of basjoo flower extract, 40 parts of stem cell extract, 10 parts of camellia oil, 5 parts of nicotinamide, 5 parts of vitamin C, 2 parts of panthenol, 3 parts of PEG-40 hydrogenated castor oil, 2 parts of preservative, 0.7 part of hydroxyethyl cellulose and 12 parts of deionized water.
3. The skin care article of claim 1 or 2, wherein the skin care article further comprises citric acid or sodium citrate; the stem cell extract comprises the mesenchymal stem cell exosomes and stem cell culture supernatant after being crushed.
4. A method of making a skin care article according to any of claims 1 to 3 comprising the steps of:
(1) weighing the raw materials according to the formula;
(2) putting nicotinamide and vitamin C into a container, adding deionized water to fully dissolve, adding the basjoa flower extract, and stirring and mixing uniformly;
(3) adding hydroxyethyl cellulose into the solution obtained in the step (2), fully dissolving and stirring until the mixture is homogeneous;
(4) adding camellia oil, panthenol and PEG-40 hydrogenated castor oil into the solution obtained in the step (3), and stirring to homogenize;
(5) adding the stem cell extract into the solution obtained in the step (4), stirring until the stem cell extract is uniformly mixed, then adding the preservative, and stirring and uniformly mixing;
(6) and (5) adding citric acid or sodium citrate into the solution obtained in the step (5) to adjust the pH value, thus obtaining the skin care product.
5. The method of claim 4, wherein the basho extract is prepared by a method comprising: extracting fresh flos Bombacis Cannabifolii under reflux for 3 times; extracting flos Bombacis Cannabinae and 65% ethanol at a mass ratio of 1:18 for 1.5h at the 1 st time; extracting flos Bombacis Cannabinae and 60% ethanol at a mass ratio of 1:10 for 1.5h for the second time; extracting flos Bombacis Cannabinae and 60% ethanol at a mass ratio of 1:8 for 1.5h at the 3 rd time; filtering the extractive solutions extracted for 3 times with three layers of gauze, mixing filtrates, and concentrating the filtrate under reduced pressure to 5 times of the weight of flos Bombacis Cannabinae to obtain the final product.
6. The method of claim 4, wherein the preservative is prepared by: weighing 6 parts of methyl p-hydroxybenzoate, adding a 17-time volume of 1, 3-butanediol solution, stirring until the solution is clear, and fully dissolving to obtain the preservative; the pH value in the step (6) is 6.
7. The method of claim 4, wherein the stem cell extract is prepared by the method comprising the steps of:
(1) carrying out in-vitro large-scale culture on stem cells: culturing according to GMP cell to obtain mesenchymal stem cell;
(2) carrying out ultrasonic crushing on the mesenchymal stem cells collected after the culture in the step (1) to obtain a crushed mesenchymal stem cell exosome;
(3) collecting the culture supernatant of the mesenchymal stem cells in the step (1) to obtain a stem cell culture supernatant exosome;
(4) and (3) respectively dissolving the crushed mesenchymal stem cell exosomes in the step (2) and the stem cell culture supernatant exosomes in the step (3) in normal saline, mixing in equal volume, and stirring uniformly to obtain the stem cell extract.
8. The method according to claim 7, wherein in the step (1), the mesenchymal stem cells are expanded by using ten layers of GMP-grade equipment in a cell factory, and the mesenchymal stem cells are harvested to obtain 3 x 10 cells when the cell fusion rate is greater than 95%8The mesenchymal stem cell of (a);
in the step (2), the mesenchymal stem cells are subjected to ultrasonic disruption in a specific operation of 1 × 108Adding the cell number into 100ml of physiological saline, centrifuging at 4 ℃ for 10min at 300 Xg to remove cell debris, and collecting the supernatant; however, the device is not suitable for use in a kitchenCentrifuging at 4 deg.C 2500 Xg for 30min to remove apoptotic bodies, and collecting supernatant; centrifuging at 13000 Xg for 1h to remove platelet impurities, and collecting the supernatant; centrifuging at 4 deg.C 100000 × g for 70min to obtain the crushed exosome of mesenchymal stem cell;
in the step (3), the specific operation of collecting the mesenchymal stem cell culture supernatant is as follows: centrifuging the mesenchymal stem cells in the step (1) at the temperature of 4 ℃ of 2000 Xg for 10min to remove cell debris, and collecting supernatant; centrifuging at 13000 Xg for 1h to remove platelet impurities, and collecting the supernatant; centrifuging at 4 deg.C 100000 × g for 70min to obtain exosome of stem cell culture supernatant;
in the step (4), the protein concentration of the stem cell extract is 40 mg/ml.
9. The method for preparing a recombinant protein of claim 7 or 8, wherein the disrupted mesenchymal stem cell exosomes of step (2) and the stem cell culture supernatant exosomes of step (3) are subjected to BCA protein concentration determination, respectively, wherein the protein concentration determined in step (2) is 9.5mg/ml, and the protein concentration determined in step (3) is 53.6 mg/ml.
10. Use of a skin care product according to any one of claims 1 to 3 in the manufacture of a stretch mark removal product.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910938897.5A CN110652491B (en) | 2019-09-30 | 2019-09-30 | Skin care product with effect of removing striae gravidarum and preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910938897.5A CN110652491B (en) | 2019-09-30 | 2019-09-30 | Skin care product with effect of removing striae gravidarum and preparation method and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN110652491A true CN110652491A (en) | 2020-01-07 |
| CN110652491B CN110652491B (en) | 2021-09-21 |
Family
ID=69040174
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910938897.5A Active CN110652491B (en) | 2019-09-30 | 2019-09-30 | Skin care product with effect of removing striae gravidarum and preparation method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110652491B (en) |
Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103405377A (en) * | 2013-08-15 | 2013-11-27 | 江南大学 | Natural skin whitening and protecting composition and application thereof in cosmetics |
| CN105267243A (en) * | 2014-07-23 | 2016-01-27 | 深圳天元干细胞科技有限公司 | Stem cell extract product for eliminating skin striae gravidarum |
| CN105662949A (en) * | 2016-01-19 | 2016-06-15 | 佛山市芊茹化妆品有限公司 | Cosmetic composition with whitening function, method for preparing cosmetic composition and application thereof |
| CN107753410A (en) * | 2017-12-06 | 2018-03-06 | 广州赛莱拉干细胞科技股份有限公司 | Striae gravidarum repair paste containing epidermal stem cell extract |
| CN107802588A (en) * | 2017-12-04 | 2018-03-16 | 广州赛莱拉干细胞科技股份有限公司 | A kind of skin care compositions of the anti-aging of cell extract containing raisins and application |
| CN109745618A (en) * | 2017-11-07 | 2019-05-14 | 陕西博鸿生物科技集团有限公司 | A kind of restorative procedure of striae of pregnancy |
| CN110075146A (en) * | 2019-06-13 | 2019-08-02 | 广州赛琅生物技术有限公司 | A kind of canker sore spray and preparation method thereof |
| CN110075050A (en) * | 2019-03-14 | 2019-08-02 | 江苏全能干细胞生物工程有限公司 | The preparation method of cell factor freeze-dried powder in a kind of mescenchymal stem cell culture supernatant for eliminating striae of pregnancy |
-
2019
- 2019-09-30 CN CN201910938897.5A patent/CN110652491B/en active Active
Patent Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103405377A (en) * | 2013-08-15 | 2013-11-27 | 江南大学 | Natural skin whitening and protecting composition and application thereof in cosmetics |
| CN105267243A (en) * | 2014-07-23 | 2016-01-27 | 深圳天元干细胞科技有限公司 | Stem cell extract product for eliminating skin striae gravidarum |
| CN105662949A (en) * | 2016-01-19 | 2016-06-15 | 佛山市芊茹化妆品有限公司 | Cosmetic composition with whitening function, method for preparing cosmetic composition and application thereof |
| CN109745618A (en) * | 2017-11-07 | 2019-05-14 | 陕西博鸿生物科技集团有限公司 | A kind of restorative procedure of striae of pregnancy |
| CN107802588A (en) * | 2017-12-04 | 2018-03-16 | 广州赛莱拉干细胞科技股份有限公司 | A kind of skin care compositions of the anti-aging of cell extract containing raisins and application |
| CN107753410A (en) * | 2017-12-06 | 2018-03-06 | 广州赛莱拉干细胞科技股份有限公司 | Striae gravidarum repair paste containing epidermal stem cell extract |
| CN110075050A (en) * | 2019-03-14 | 2019-08-02 | 江苏全能干细胞生物工程有限公司 | The preparation method of cell factor freeze-dried powder in a kind of mescenchymal stem cell culture supernatant for eliminating striae of pregnancy |
| CN110075146A (en) * | 2019-06-13 | 2019-08-02 | 广州赛琅生物技术有限公司 | A kind of canker sore spray and preparation method thereof |
Non-Patent Citations (2)
| Title |
|---|
| 徐洁等: "《芳香植物研究与应用》", 31 October 2017, 云南科技出版社 * |
| 胡小毅: "《佛道教与养生》", 30 April 2005, 中国物资出版社 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN110652491B (en) | 2021-09-21 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN110731924B (en) | Enhanced anti-aging cosmetic compositions | |
| CN109157451B (en) | Acne-removing facial cleanser | |
| CN110664644B (en) | Skin care composition containing folic acid, essence and preparation method thereof | |
| CN111388346A (en) | Freeze-dried powder live mask with vegetable protein and polypeptide formula and preparation method thereof | |
| CN106420459B (en) | A kind of ginseng water Essence living and preparation method thereof and the facial mask comprising it | |
| CN112402347A (en) | Acne-removing repair composition containing fibronectin and preparation method thereof | |
| CN106420491A (en) | Multifunctional skin care product with effects of whitening skin, removing freckles, preserving moisture and removing wrinkle and preparation method of multifunctional skin care product | |
| CN110559230A (en) | Anti-aging and anti-wrinkle essence lotion and preparation method thereof | |
| CN113384482A (en) | Moisturizing and skin-tendering essence and preparation method thereof | |
| CN110652491B (en) | Skin care product with effect of removing striae gravidarum and preparation method and application thereof | |
| TW202106323A (en) | Use ofrosmarinus officinaliscallus extract for manufacturing composition inhibiting skin aging and a culture medium for inducingrosmarinus officinaliscallus | |
| TW201536343A (en) | Composition for skin anti-ageing treatment | |
| KR20200132661A (en) | Composition for improving atopy dermatitis, Atopic dermatitis remedy and Manufacturing method thereof | |
| CN110101643A (en) | A kind of moisturizing remediation composition and the cosmetics containing the composition | |
| CN109568230A (en) | A kind of Chinese medicine cosmetic of the anti-aging of moisturizing whitening | |
| CN107028830B (en) | Medicine for preventing scar hyperplasia and preparation process thereof | |
| CN113171316B (en) | Cosmetic for repairing and relieving skin irritation and preparation method thereof | |
| CN115337233A (en) | Gel mask with mild acne removing effect and preparation method thereof | |
| CN114159341A (en) | Freeze-dried eye patch and preparation method thereof | |
| CN114469811A (en) | Collagen anti-wrinkle repair mask and production process | |
| CN114404349A (en) | Calf blood and centella asiatica derived exosome composition and application thereof in skin repair | |
| CN113274339A (en) | Silicone oil cream with itching relieving and moisture keeping effects and preparation method thereof | |
| CN109010105B (en) | Multi-effect composition and preparation method and application thereof | |
| CN110946770A (en) | Acne-removing cream with liquid crystal structure and preparation method thereof | |
| CN113384483A (en) | Composition for improving skin elasticity and delaying aging and preparation method thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |