CN111714396A - Composition containing ectoin and application thereof in laser beauty treatment - Google Patents
Composition containing ectoin and application thereof in laser beauty treatment Download PDFInfo
- Publication number
- CN111714396A CN111714396A CN202010711868.8A CN202010711868A CN111714396A CN 111714396 A CN111714396 A CN 111714396A CN 202010711868 A CN202010711868 A CN 202010711868A CN 111714396 A CN111714396 A CN 111714396A
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- sodium hyaluronate
- ectoin
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Classifications
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/49—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds
- A61K8/494—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds with more than one nitrogen as the only hetero atom
- A61K8/4953—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds with more than one nitrogen as the only hetero atom containing pyrimidine ring derivatives, e.g. minoxidil
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
- A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/715—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
- A61K31/726—Glycosaminoglycans, i.e. mucopolysaccharides
- A61K31/728—Hyaluronic acid
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/33—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing oxygen
- A61K8/34—Alcohols
- A61K8/345—Alcohols containing more than one hydroxy group
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- 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
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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
- A61K8/72—Cosmetics or similar toiletry preparations characterised by the composition containing organic macromolecular compounds
- A61K8/73—Polysaccharides
- A61K8/735—Mucopolysaccharides, e.g. hyaluronic acid; Derivatives thereof
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
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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
-
- 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
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/40—Chemical, physico-chemical or functional or structural properties of particular ingredients
- A61K2800/59—Mixtures
- A61K2800/592—Mixtures of compounds complementing their respective functions
Abstract
The invention relates to an application of a composition containing ectoin in laser beauty treatment, and the invention comprises the following components in percentage by weight: 2-3 parts of ectoin, 0.1-0.5 part of high-molecular sodium hyaluronate, 0.1-0.5 part of oligomeric sodium hyaluronate, 2-9 parts of sodium dermatan sulfate, 3-5 parts of butanediol and water for supplementing 100 parts by weight. The obtained composition can be used for synergistic effect, and preventing and repairing complications of laser therapy such as various inflammatory reactions. The composition obtained by the invention is moist and non-irritant, has no toxic or side effect, is simple in preparation method and easy to compound, and can be applied to pharmaceutical preparations, cosmetics and the like.
Description
Technical Field
The invention belongs to the technical field of cosmetics, and particularly relates to a composition containing ectoin and application of preventive and reparative anti-inflammatory effects of the composition in laser cosmetology.
Background
Nowadays, the pursuit of people for beauty is continuously promoted, the technology in the field of beauty is continuously improved, and laser beauty is one of the effects of beauty treatment. Laser therapy relies on the thermal action of high-energy light waves to selectively and effectively destroy target cells and avoid damage to surrounding normal tissues, and is currently used in the fields of pigment pathological changes, wrinkle removal, photon skin tendering and the like. Although laser beauty has the advantages of good curative effect, less side effect, high safety and the like compared with the prior beauty technology, the laser beauty still needs postoperative care and has the defects of postoperative complications.
Postoperative complications are mainly inflammatory reactions including edema, exudation, scabbing, redness, erythema, etc. Redness of the skin results from vasodilation, which tends to produce pigmentation, which generally resolves longer and is more difficult for yellow people to resolve than for white people. Erythema occurs mostly during the healing process of wounds, and if erythema is too hard or healed too long, hypertrophic scarring may occur. Most of the existing treatment methods are to repair laser beauty complications by applying or taking medicines after operation, such as (1) dexamethasone is orally taken after operation to relieve swelling; (2) biological synthetic semitransparent dressing or topical antibiotic ointment is used for absorbing wound surface exudate and moisturizing, and healing is accelerated; (3) after operation, the vitamin C synthetic cream or sunscreen cream, hydroquinone cream and moisturizing cream are applied for sunscreen and moisturizing. But few options exist for pre-operative prophylaxis.
According to literature reports (Simon et al 1995), Hsp70 overexpression can significantly reduce the release of IL-6 induced by UV-A, UV-B and oxidative stress, thereby performing cell protection and repair, and protecting cells from infection, inflammation or various injuries. In addition, ectoin has the effect of inducing the over-expression of Hsp70 gene (Buomino et al 2005), and after the addition of thermal stimulation to cells treated by ectoin, experimental results show that the synthesis of Hsp70 of human keratinocytes is increased, and proinflammatory cytokines IL-1a, IL-6, IL-8 and TNF-alpha are down-regulated, thereby showing that the ectoin has anti-inflammatory effect.
Aiming at the characteristics that the ectoin promotes the overexpression of Hsp70 so as to obviously reduce inflammation and resist heat action and the like, the invention prepares a mixture containing the ectoin, which can be used before and after laser treatment and furthest reduce complication symptoms in the aspects of diminishing inflammation, removing swelling, easing pain and preserving moisture. Publication No. CN110215461A discloses a medical cosmetic formula, which mainly comprises hyaluronic acid, polylysine and vitamin C; publication No. CN104474577B also discloses a medical beauty formula, which mainly comprises hyaluronic acid, yeast essence and bright peptide antibacterial liquid, but the medical beauty formula is only used for skin repair after operation and carries out repair and nursing on inflammation and wounds which have already occurred. Publication No. CN106420399A discloses a mixed reagent, wherein a nursing agent 1 is used in preoperative operation, a nursing agent 2 is used in postoperative operation, most of the components of the nursing agent 1 with a prevention function are humectants, an anti-inflammatory effect substance is glycyrrhetinic acid, and prevention effect data are not given in the examples. Publication No. CN105477342A discloses a traditional Chinese medicine repair cream for post-operative whitening repair, which contains various traditional Chinese medicine extracts such as centella asiatica and ligusticum wallichii, but the natural plant extracts contain unknown side effects or adverse reactions under synergistic effect. The publication No. CN110141627A discloses a formula for wound care, which comprises aloe gel, propolis and the like, but has the effects of sterilization and pain relief, has no obvious anti-inflammatory effect and is also used for postoperative care. Thus, there is a need to develop a formulation that can be used to prevent laser cosmetic complications.
Disclosure of Invention
In order to achieve the above object, the present invention provides a composition containing ectoin, which can preventively and restoratively alleviate skin inflammation, calm down and remove heat, and can be applied to laser beauty treatment. The following technical scheme is adopted.
1. A composition comprising ectoin, polymeric sodium hyaluronate, oligomeric sodium hyaluronate, sodium dermatan sulfate, butylene glycol, and purified water.
2. The composition according to item 1, wherein the ectoin is 2 to 3 parts by weight, preferably 2.5 parts by weight, based on 100 parts by weight of the total composition.
3. The composition according to item 1 or 2, wherein the amount of the polymeric sodium hyaluronate is 0.1 to 0.5 parts by weight, preferably 0.3 parts by weight, based on 100 parts by weight of the total composition.
4. The composition as claimed in any one of claims 1 to 3, wherein the molecular weight of the polymeric sodium hyaluronate is 100 to 180 ten thousand daltons.
5. The composition according to any one of items 1 to 4, wherein the oligomeric sodium hyaluronate is 0.1 to 0.5 parts by weight, preferably 0.5 parts by weight, based on 100 parts by weight of the total composition.
6. The composition as claimed in any one of claims 1 to 5, wherein the oligomeric sodium hyaluronate has a molecular weight of less than 10000 Dalton.
7. The composition according to any one of items 1 to 6, wherein the dermatan sulfate is 2 to 9 parts by weight, preferably 5 parts by weight, based on 100 parts by weight of the total composition.
8. A composition according to any one of claims 1 to 7, wherein the dermatan sulfate sodium has a molecular weight of 30000 to 60000 daltons.
9. The composition according to any one of items 1 to 8, wherein the butanediol is 3 to 5 parts by weight, preferably 5 parts by weight, based on 100 parts by weight of the total composition.
10. The composition according to any one of items 1 to 9, further comprising a cosmetically and medically acceptable carrier and auxiliary materials.
11. A cosmetic composition comprising the composition according to any one of items 1 to 10.
12. The cosmetic according to claim 11, wherein the cosmetic comprises a toner, a spray, a lotion, a cream, an eye cream, essence, massage oil, body lotion, or the like.
13. Use of the cosmetic according to item 11 or 12 as a component for preventing laser cosmetic complications and repairing.
14. A pharmaceutical preparation comprising the composition according to any one of claims 1 to 10.
15. The pharmaceutical formulation according to claim 14, wherein the pharmaceutical formulation comprises an ointment, liniment, gel, aerosol, patch, etc. for skin.
16. Use of the pharmaceutical preparation according to item 14 or 15 as a component for the prevention of laser cosmetic complications and for the repair.
The composition disclosed by the invention is a combination of the ectoin, the hyaluronic acid substance and the dermatan sulfate sodium. The ectoin has multiple effects, can repair laser beauty injury or potential injury parts from the aspects of diminishing inflammation, moisturizing, sun protection and the like, can perform synergistic interaction by combining with high-molecular sodium hyaluronate, oligomeric sodium hyaluronate and dermatan sulfate sodium, can exert respective functions to the maximum extent, and can preventively eliminate complications such as inflammatory pain and the like before laser beauty operation; after operation, the composition still has the effects of anti-inflammation repair and the like, and can continuously cure complications. Meanwhile, the composition also has the effects of moisturizing and whitening the skin and the like, can be continuously used after operation, and helps to protect and maintain the skin after the concurrent symptoms are relieved.
Detailed description of the invention
The application provides a composition for preventing laser cosmetic complications and repairing, which comprises ectoin, high-molecular sodium hyaluronate, oligomeric sodium hyaluronate, sodium dermatan sulfate and butanediol.
Ectoin (Ectoin) is a natural protective component which is produced by desert halophilic bacteria under the environment of high temperature, dryness, strong ultraviolet irradiation and high salinity, is positioned on the outer layer of cells, can maintain the osmotic pressure balance inside and outside the cells, and can prevent the cells from dehydrating. The ectoine is a compatible substance which is most widely distributed in the bacterial community and is compatible with metabolism in cells, and does not influence the biomacromolecule function or physiological process of the cells. Ectoin also has other important effects: the surface charge of the molecule is dense, and water molecules are easy to combine, so that the moisturizing effect is achieved; can inhibit melanin synthesis, and has skin whitening effect.
In a specific embodiment of the present application, the ectoin is 2 to 3 parts by weight, for example, 2, 2.5, 3 parts by weight, and more preferably 2.5 parts by weight, based on 100 parts by weight of the total composition.
Hyaluronic acid is widely present in intercellular substance in a living body, has no species difference and immunogenicity, has good biodegradability and biocompatibility, and is an ideal natural moisturizing factor.
The high molecular sodium hyaluronate has excellent film forming property and lubricating property, forms a hydrated film on the surface of skin, effectively keeps moisture in stratum corneum, prevents internal moisture from evaporating and isolates external bacterial dust and the like from invading.
In a specific embodiment of the present application, the molecular weight of the polymeric sodium hyaluronate is 100-180 ten thousand daltons. For example, the polymeric sodium hyaluronate of the present application may have a molecular weight of 100, 110, 120, 130, 140, 150, 160, 170, 180 kilodaltons.
In a specific embodiment of the present application, the amount of the polymeric sodium hyaluronate is 0.1 to 0.5 parts by weight, for example, 0.1, 0.2, 0.3, 0.4, 0.5 parts by weight, based on 100 parts by weight of the total composition.
The oligomeric sodium hyaluronate is sodium hyaluronate with higher bioactivity and molecular weight lower than 10kDa, can be absorbed through skin, can remove free radicals, and has the effects of repairing cells, promoting growth, deeply preserving moisture and repairing after drying in the sun.
In a specific embodiment of the present application, the oligomeric sodium hyaluronate is present in an amount of 0.1 to 0.5 parts by weight, for example 0.1, 0.2, 0.3, 0.4, 0.5 parts by weight, and more preferably 0.5 parts by weight, based on 100 parts by weight of the total composition. The molecular weight is less than 10000 Dalton.
In a specific embodiment of the present application, dermatan sulfate sodium and butylene glycol are further included. The dermatan sulfate sodium is glycosaminoglycan widely existing in mammalian bodies, has an inhibitory effect on cell degranulation, can resist inflammation, and has a certain analgesic effect. Butanediol is also a cosmetic moisturizer. In one embodiment, the dermatan sulfate sodium is 2 to 9 parts by weight, preferably 5 parts by weight, relative to 100 parts by weight of the total composition, and has a molecular weight of 30000 to 60000 daltons; the butanediol accounts for 3-5 parts by weight, and preferably 5 parts by weight relative to 100 parts by weight of the total weight of the composition.
In a particular embodiment of the present application, the composition further comprises a cosmetically or medically acceptable carrier or adjuvant. The acceptable excipients include, but are not limited to, solvents, solubilizers, preservatives, antioxidants, pH adjusting agents, penetration enhancers, liposomes, humectants, thickeners, chelating agents, skin feel modifiers, surfactants, emulsifiers, propellants/propellants, fragrances, and pigments.
The present application also provides a cosmetic having the composition. Such cosmetics include, but are not limited to, soaps, facial cleansers, toners, sprays, lotions, creams, eye creams, essences, shower gels, massage oils, body lotions, facial masks, and the like.
The present application also provides pharmaceutical formulations having the compositions. The pharmaceutical preparation includes but is not limited to ointment, liniment, gel, aerosol, patch and other moisturizing and anti-inflammatory drugs for skin, ophthalmology or nasal cavity.
The ectoin in the composition, the cosmetic and the pharmaceutical preparation provided by the application has multiple effects, and can repair laser beauty damage or potential damage parts from the aspects of diminishing inflammation, moisturizing, preventing sun and the like. In addition, the ectoin is compounded with the high-molecular sodium hyaluronate and the oligomeric sodium hyaluronate, the three substances are synergistic, respective functions can be exerted to a greater extent, the prevention effect of the composition on laser injury is improved, the effects of diminishing inflammation and preserving moisture can be exerted before laser cosmetic surgery, and compared with the traditional method that only postoperative repair is adopted, a protective barrier can be formed in advance, and the skin can be protected more effectively and perfectly.
The following examples of the present application are intended only to illustrate specific embodiments for carrying out the present application and these embodiments are not to be construed as limiting the present application. Other changes, modifications, substitutions, combinations, and simplifications which may be made without departing from the spirit and principles of the present application are intended to be equivalent substitutions and are within the scope of the present application.
Examples
The experimental methods used in the following examples are all conventional methods, unless otherwise specified.
The ectoin used in the following examples was purchased from sigma, polymeric sodium hyaluronate from Huaxi Biotech Co., Ltd, oligomeric sodium hyaluronate from Huaxi Biotech Co., Ltd, sodium dermatan sulfate from China pharmaceutical and biological products institute, and butanediol from China pharmaceutical group.
Experimental example 1: composition for preventing and repairing light injury
The damage caused to the skin by ultraviolet laser light is mainly photodamage. In the experiment, before ultraviolet irradiation, the immortalized human keratinocyte cell line is treated by a sample, and the cell proliferation condition is observed, so that whether the composition has a certain effect of preventing photodamage or not is determined. The experiment also carries out sample treatment on the cells after illumination, and observes whether the cells have certain repairing and anti-inflammatory effects.
1. Experimental materials and instruments including ectoin (sigma, tetrahydropyrimidine) and high molecular sodium hyaluronate (molecular weight 10 × 10)5-18×105Da, Huaxi Biotech Ltd.), oligomeric sodium hyaluronate (molecular weight < 10 × 103Da, Huaxi Biotech Co., Ltd.), dermatan sulfate sodium (3 × 104-6×104Da, institute for drug and biological products, china), butanediol (national drug group), immortalized human keratinocyte cell line (HaCaT cell, shanghai enzyme-linked biotechnology limited), mtt (sigma), DMEM medium (GibcoBRL, usa)) Fetal bovine serum (GibcoBRL, usa), EDTA (national group), DMSO (national group), pancreatin (GibcoBRL, usa), PBS (GibcoBRL, usa), 96-well cell culture plates (serela fly); an ultra clean bench (Beijing Dongherhaar instruments manufacturing Co., Ltd., SCB-1520), an enzyme linked immunoassay analyzer (Bio-Rad, USA), a UV-8 type ultraviolet light box (Beijing electric light source research institute), a ST-513 type ultraviolet measuring instrument (Taiwan Chichi), a carbon dioxide incubator (SANYO, MCO-18AIC), a microscope (OLYMPUS, CKX31), and a constant temperature water bath (Daorkun instruments factory in Jintani city).
2. The experimental method comprises the following steps:
2.1 solution preparation:
sample group composition solution preparation: according to the content of each component shown in table 1, corresponding ectoin, high-molecular sodium hyaluronate, oligomeric sodium hyaluronate and dermatan sulfate sodium are added into butanediol to be dissolved and uniformly mixed until the mixture is clear, water is added to make up 100%, and composition solutions of each sample group are prepared, wherein the composition solutions comprise comparative examples 1-7 and examples 1-3. The composition is shown in table 1:
TABLE 1
Preparing a culture medium solution: taking 0.01g of each sample group composition solution in the table 1 respectively, dissolving in 1mL of culture medium respectively, filtering, sterilizing, and storing at 4 ℃;
preparing an MTT solution: MTT solution (10mL PBS dissolved 50mg MTT dry powder) with concentration of 5mg/mL is prepared by PBS, filtered and sterilized, and stored in dark at 4 ℃.
Preparing a digestive juice: 0.02% EDTA solution: 20mg of EDTA (disodium edetate) is weighed and dissolved in 100ml of D-Hanks solution, filtered and sterilized by a 0.22 mu m disposable filter, and stored at 4 ℃. 0.25% pancreatin solution: weighing 250mg of trypsin, dissolving in 100ml of D-Hanks solution, filtering and sterilizing with a 0.22 μm disposable filter, and storing at 4 ℃.
2.2 cell culture: HaCaT cells were cultured at 37 ℃ with 5% CO2HaCaT cells were cultured in medium containing 10% fetal bovine serum in a conditioned cell culture chamber. 0.02% EDTA and 0.25% trypsin were digested for passage and plated in petri dishes or plates for further experiments.
2.3 prevention of photodamage experiment with composition:
firstly, 0.02% EDTA and 0.25% pancreatin are used for carrying out enzymolysis digestion on HaCaT cells in a logarithmic growth phase in a sub-fusion state for 5-10 min, 100 mu L of HaCaT cells are inoculated to a 96-hole cell culture plate, and the concentration of suspension cells is adjusted to be 4 × 104/mL,37℃、5%CO2The culture was routinely carried out overnight. After overnight incubation, the culture medium was aspirated, washed once with PBS, and 100. mu.L of each prepared composition solution of each sample group of Table 1 was added. Control group 1 and control group 2 were separately prepared, and 100. mu.L of fresh culture medium was added. All are put into a cell culture box to be incubated for 24 h. The post-treatment method comprises the following steps:
the control group 1 was not irradiated with ultraviolet light, and the fresh culture solution was replaced;
irradiating the control group 2 with ultraviolet light, and then replacing the fresh culture solution;
each sample set was irradiated with uv light, after which the fresh culture medium was replaced.
Ultraviolet light conditions: 30J/cm2UVA and 90mJ/cm2UVB. (long-wave and short-wave ultraviolet light waves are mostly used in laser cosmetology), after irradiation/non-irradiation, 20 μ L of MTT with a concentration of 5mg/ml is added to each 100 μ L of culture medium, incubation is performed for 4 hours, supernatant is discarded, an equal amount of dimethyl sulfoxide (DMSO) is added, shaking is performed at room temperature for 15 minutes, optical density value (OD value) of each well is measured at 490nm wavelength by a microplate reader, damage rate is (non-irradiated group OD)/non-irradiated group OD × 100%
2.4 Effect test of the composition on repairing photodamage:
first, HaCaT cells in the logarithmic growth phase of the sub-confluent state were subjected to enzymatic digestion with 0.02% EDTA and 0.25% pancreatin, and 100. mu.L of the cells were inoculated into a 96-well cell culture plate, and the suspension cell concentration was adjusted to 4 × 104/mL,37℃、5%CO2The culture was routinely carried out overnight. After overnight incubation, the culture medium was aspirated and washed with PBSOne pass. The post-treatment method comprises the following steps:
the control group 1 was replaced with 100. mu.L of fresh culture medium without irradiation of ultraviolet light;
control group 2 was irradiated with ultraviolet light, and then 100. mu.L of fresh culture medium was replaced;
each sample set was irradiated with ultraviolet light, after which 100 μ L of each sample set solution was replaced.
And finally, putting the mixture into a cell culture box to incubate for 24 h. The ultraviolet condition and the MTT cell activity test method are the same as above.
3. The experimental results are as follows:
the experimental results of the composition for preventing and repairing the light damage are shown in table 2.
TABLE 2
Grouping | Prevention of light absorption in experiments | The damage rate% | Light absorption value of repair experiment | The damage rate% |
Control group 1 | 0.99±0.03 | 0.00 | 1.04±0.02 | 0.00 |
Control group 2 | 0.30±0.04 | 69.70 | 0.32±0.02 | 69.23 |
Blank control | 0.31±0.03 | 68.69 | 0.31±0.00 | 70.19 |
Example 1 | 0.88±0.04 | 11.11 | 0.86±0.02 | 17.12 |
Example 2 | 0.94±0.04 | 5.05 | 0.91±0.01 | 12.50 |
Example 3 | 0.97±0.03 | 2.02 | 0.97±0.01 | 6.92 |
Comparative example 1 | 0.64±0.01 | 35.35 | 0.63±0.01 | 39.81 |
Comparative example 2 | 0.66±0.04 | 33.33 | 0.67±0.01 | 35.96 |
Comparative example 3 | 0.77±0.00 | 22.22 | 0.68±0.05 | 34.59 |
Comparative example 4 | 0.85±0.02 | 14.14 | 0.76±0.03 | 26.77 |
Comparative example 5 | 0.55±0.06 | 44.44 | 0.48±0.04 | 53.85 |
Comparative example 6 | 0.81±0.01 | 18.18 | 0.72±0.01 | 31.19 |
Comparative example 7 | 0.31±0.05 | 68.69 | 0.38±0.00 | 63.45 |
The photodamage pretreatment experiment shows that the main factor influencing the proliferation activity of human keratinocytes is ectoine, oligomeric sodium hyaluronate is inferior, and the macromolecular sodium hyaluronate has a certain effect, but has a poor effect compared with the oligomeric sodium hyaluronate and a tiny effect of sodium dermatan sulfate; the synergistic effect of the first three substances is greater than the effect of compounding the ectoin with one hyaluronic acid, and the synergistic effect of the dermatan sulfate is small. When the concentration of ectoin is 2.5 percent and the concentration of oligomeric sodium hyaluronate is 0.5 percent, the effect is best, the damage effect is greatly reduced, and the composition has a good effect of preventing laser light damage. In a repairability experiment, the four components have good effects and synergistic effects, and the composition has good anti-inflammatory and repairing effects on laser photodamage.
Experimental example 2 prevention and repair effects of the composition on thermal injury
The damage to the skin by the infrared laser is mainly thermal damage. In this experiment, the immortalized human keratinocyte cell line was treated with a sample before heating, and the cell proliferation was observed, thereby determining whether the composition had a certain effect of preventing thermal injury. The experiment also carries out sample treatment on the heated cells, and observes whether the cells have certain repairing and anti-inflammatory effects.
1. Experimental materials and instruments including ectoin (sigma, tetrahydropyrimidine) and high molecular sodium hyaluronate (molecular weight 10 × 10)5-18×105Da, Huaxi Biotech Ltd.), oligomeric sodium hyaluronate (molecular weight < 10 × 103Da, Huaxi Biotech Co., Ltd.), dermatan sulfate sodium (3 × 104-6×104Da, chinese institute for drug and biological products), butylene glycol (national group of pharmaceuticals), immortalized human keratinocyte cell line (HaCaT cells, shanghai enzyme-linked biotechnology limited), DMEM medium (GibcoBRL, usa), fetal bovine serum (GibcoBRL, usa), EDTA (national group of pharmaceuticals), pancreatin (GibcoBRL, usa), PBS (GibcoBRL, usa), 96-well cell culture plate (semer fly), human IL-6ELISA kit (usa R)&D Systems corporation); an ultra clean bench (Beijing Donghong Harr instruments manufacturing Co., Ltd., SCB-1520), an enzyme linked immunoassay analyzer (Bio-Rad, USA), a carbon dioxide incubator (SANYO, MCO-18AIC), a microscope (OLYMPUS, CKX31), and a constant temperature water bath (Tan Dai instruments factory).
2. The experimental method comprises the following steps:
2.1 solution preparation:
sample group composition solutions, medium solutions, and the like were prepared in the method of 2.1 in experimental example 1.
2.2 cell culture: cell culture was performed in the manner of 2.2 in example 1.
2.3 preventive effect of composition on thermal injury test:
first, HaCaT cells in the logarithmic growth phase of the sub-confluent state were subjected to enzymatic digestion with 0.02% EDTA and 0.25% pancreatin, and 100. mu.L of the cells were inoculated into a 96-well cell culture plate, and the suspension cell concentration was adjusted to 4 × 104/mL,37℃、5%CO2The culture was routinely carried out overnight.
To the cell fluid, 100. mu.L of each prepared composition solution of the sample group shown in Table 1 was added, the cell culture plate was divided into a temperature control group and a heat shock group, the heat shock group was placed in a 47 ℃ water bath for 1 hour, the temperature control group was placed in a 37 ℃ water bath for 1 hour, and fresh culture solution was immediately replaced for all cell suspensions after heating. Then all return 5% CO2And continuously culturing in a constant-temperature incubator at 37 ℃. Collecting the culture solution of the cell supernatant of each group for 24 hours; sucking out the culture solution with a sterile pipette, determining that the cell activity is normal by a microscope, centrifuging at 4000r/min for 10min, subpackaging the supernatant, and storing at-70 ℃ to be tested.
The supernatant IL-6 was assayed using an ELISA test kit, following strict instructions, with fresh culture medium as negative control. The increase in IL-6 concentration (IL-6 concentration at 47 ℃ C. -IL-6 concentration at 37 ℃ C.)/IL-6 concentration at 37 ℃ C. times.100%
2.4 Effect of the composition on the repair of thermal injury:
first, HaCaT cells in the logarithmic growth phase of the sub-confluent state were subjected to enzymatic digestion with 0.02% EDTA and 0.25% pancreatin, and 100. mu.L of the cells were inoculated into a 96-well cell culture plate, and the suspension cell concentration was adjusted to 4 × 104/mL,37℃、5%CO2The culture was routinely carried out overnight.
Replacing fresh culture solution for all cell suspensions, dividing the cell culture plate into a temperature control group and a heat shock group, placing the heat shock group in a 47 deg.C water bath tank for 1h, placing the temperature control group in a 37 deg.C water bath tankFor 1 hour, 100. mu.L of each prepared composition solution of the sample group shown in Table 1 was added to the cell sap immediately after heating. Then all return 5% CO2And continuously culturing in a constant-temperature incubator at 37 ℃. Collecting the culture solution of the cell supernatant of each group for 24 hours; sucking out the culture solution with a sterile pipette, determining that the cell activity is normal by a microscope, centrifuging at 4000r/min for 10min, subpackaging the supernatant, and storing at-70 ℃ to be tested.
The supernatant IL-6 was assayed using an ELISA test kit, following strict instructions, with fresh culture medium as negative control.
3. The experimental results are as follows:
the concentration of IL-6 protein (pg/mL) in the supernatant after heat damage pretreatment of HaCaT cells is shown in Table 3; the concentration of IL-6 protein (pg/mL) in the supernatant after heat damage post-treatment of HaCaT cells is shown in Table 4.
TABLE 3
Grouping | 37℃ | 47℃ | The growth rate% |
Blank control | 202.73±20.10 | 516.39±60.47 | 154.72 |
Example 1 | 200.56±22.39 | 371.78±50.99 | 85.37 |
Example 2 | 195.94±9.18 | 353.77±28.03 | 80.55 |
Example 3 | 196.56±24.01 | 340.42±45.72 | 73.19 |
Comparative example 1 | 196.78±11.61 | 396.81±30.36 | 101.65 |
Comparative example 2 | 195.21±13.40 | 386.85±66.51 | 98.17 |
Comparative example 3 | 194.74±11.62 | 384.77±77.05 | 97.58 |
Comparative example 4 | 195.41±24.94 | 365.69±33.22 | 87.14 |
Comparative example 5 | 202.92±32.97 | 411.28±47.38 | 102.68 |
Comparative example 6 | 204.16±7.64 | 395.52±36.09 | 93.73 |
Comparative example 7 | 203.11±8.88 | 496.06±40.64 | 144.23 |
TABLE 4
Grouping | 37℃ | 47℃ | The growth rate% |
Blank control | 203.65±20.10 | 520.14±56.27 | 155.41 |
Example 1 | 200.64±13.34 | 311.91±33.11 | 55.46 |
Example 2 | 197.60±10.79 | 294.74±52.41 | 49.16 |
Example 3 | 198.00±17.63 | 290.66±52.95 | 46.80 |
Comparative example 1 | 198.91±12.44 | 419.20±55.22 | 110.75 |
Comparative example 2 | 201.45±9.70 | 396.90±60.44 | 97.02 |
Comparative example 3 | 201.73±12.12 | 397.02±50.86 | 96.81 |
Comparative example 4 | 198.28±18.98 | 330.67±51.50 | 66.77 |
Comparative example 5 | 197.43±4.31 | 376.14±46.68 | 90.52 |
Comparative example 6 | 199.58±16.77 | 351.34±42.55 | 76.04 |
Comparative example 7 | 203.90±12.05 | 501.02±47.22 | 145.72 |
Both experiments show that the secretion amount of the proinflammatory factor IL-6 at 47 ℃ is larger than that at 37 ℃, and the IL-6 concentration at 37 ℃ is basically unchanged in each sample group due to the fact that the temperature at 37 ℃ is the normal temperature of a human body. In the thermal injury pretreatment experiment, at 47 ℃, the sample groups all produce resisting effect, the ectoin effect is most obvious, the high-molecular sodium hyaluronate and the oligomeric sodium hyaluronate have certain preventing effect, the dermatan sulfate sodium effect is small, the four substances have synergistic effect and mainly take the ectoin. When the concentration of the ectoine is 2.5%, the IL-6 concentration is the minimum, the damage effect is greatly reduced, and the ectoine has a good effect of preventing thermal damage. In a repairability experiment, the four components have good effects and synergistic effects, and the composition has an anti-inflammatory repairing effect on laser thermal injury.
Experimental example 3 preparation of spray provided herein
1. The formula is as follows: 2.5% of ectoin, 0.3% of high-molecular sodium hyaluronate, 0.5% of oligomeric sodium hyaluronate, 5% of sodium dermatan sulfate, 5% of butanediol, 3% of glycerol, 3% of propylene glycol, 2% of betaine, 0.1% of allantoin and the balance of water.
2. The method comprises the following steps: dissolving the above materials in purified water according to the proportion of the formula, and mixing well for dissolving (heating allantoin to 90 deg.C and then adding). Filtering the mixed solution, sterilizing, and adding into a spray bottle to obtain the final product.
3. The effect is as follows: the spray has good effects of preventing and repairing laser cosmetic complications.
Experimental example 4 preparation of the cream provided herein
1. Formula (relative to 100 parts by weight of face cream):
phase A: 3 parts by weight of cetearyl glucoside, 1.5 parts by weight of PEG-100 stearate, 1.5 parts by weight of cetearyl alcohol, 2.5 parts by weight of shea butter, 1.5 parts by weight of jojoba seed oil, 3 parts by weight of squalane, 2 parts by weight of caprylic/capric triglyceride, 2 parts by weight of dioctyl carbonate and 0.5 part by weight of tocopheryl acetate
Phase B: 1 part by weight of polydimethylsiloxane
And C phase: 3 parts of glycerin, 3 parts of butanediol, 0.2 part of allantoin, 0.4 part of acryloyl dimethyl ammonium taurate and 55.8 parts of water
Phase D: 2.5 parts of ectoin, 0.3 part of high-molecular sodium hyaluronate, 0.5 part of oligomeric sodium hyaluronate, 5 parts of sodium dermatan sulfate, 0.8 part of phenoxyethanol and 10 parts of water.
2. The method comprises the following steps: first, phase A was heated to 85 ℃. And fully mixing and dispersing the phase C, stirring and heating to 90 ℃. Mixing the phase A and the phase C, adding the phase B, homogenizing for a certain time, and cooling to 40 ℃. And finally, adding the phase D, uniformly stirring, and cooling to room temperature to obtain the face cream.
3. The effect is as follows: the cream has good effects of preventing and repairing laser beauty complications.
The above description is only a preferred embodiment of the present application, and is not intended to limit the present application in any way, and any simple modification, equivalent change and modification made to the above embodiment according to the technical spirit of the present application should be included in the protection scope of the present application.
Claims (10)
1. A composition comprising ectoin, polymeric sodium hyaluronate, oligomeric sodium hyaluronate, sodium dermatan sulfate, butylene glycol, and water.
2. The composition according to claim 1, wherein the ectoine is 2 to 3 parts by weight, preferably 2.5 parts by weight, based on 100 parts by weight of the total composition.
3. The composition according to claim 1 or 2, wherein the amount of the polymeric sodium hyaluronate is 0.1 to 0.5 parts by weight, preferably 0.3 parts by weight, based on 100 parts by weight of the total composition.
4. The composition as claimed in any one of claims 1 to 3, wherein the molecular weight of the polymeric sodium hyaluronate is 100-180 ten thousand daltons.
5. The composition according to any one of claims 1 to 4, wherein the oligomeric sodium hyaluronate is 0.1 to 0.5 parts by weight, preferably 0.5 parts by weight, relative to 100 parts by weight of the total composition.
6. The composition as claimed in any one of claims 1 to 5, wherein the oligomeric sodium hyaluronate has a molecular weight of less than 10000 Dalton.
7. Composition according to any one of claims 1 to 6, characterized in that the dermatan sulfate sodium is present in an amount of 2 to 9 parts by weight, preferably 5 parts by weight, relative to 100 parts by weight of the total composition.
8. A composition according to any one of claims 1 to 7, wherein the dermatan sulfate sodium has a molecular weight of 30000 to 60000 daltons.
9. Composition according to any one of claims 1 to 8, characterized in that the butanediol is present in a quantity of from 3 to 5 parts by weight, preferably 5 parts by weight, relative to 100 parts by weight of the total composition.
10. The composition according to any one of claims 1 to 9, further comprising cosmetically and medically acceptable carriers and adjuvants.
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