CN117982375A

CN117982375A - Vegetable oil composition for promoting cell proliferation and activity

Info

Publication number: CN117982375A
Application number: CN202410132734.9A
Authority: CN
Inventors: 王倩; 陆润霜; 祝乐; 赵亚
Original assignee: Shanghai Jahwa United Co Ltd
Current assignee: Shanghai Jahwa United Co Ltd
Priority date: 2024-01-30
Filing date: 2024-01-30
Publication date: 2024-05-07

Abstract

The present invention provides a vegetable oil composition for promoting cell proliferation and viability, comprising sweet wormwood oil and safflower seed oil, wherein the volume ratio of sweet wormwood oil to safflower seed oil is equal to or less than 1:100. The sweet wormwood oil is obtained by extracting sweet wormwood by a supercritical extraction method. The safflower seed oil of the present invention comprises 50-90 wt% linoleic acid. The invention also relates to application of the vegetable oil composition in preparing medicines or external preparations for skin with the function of promoting cell proliferation and activity.

Description

Vegetable oil composition for promoting cell proliferation and activity

Technical Field

The invention relates to the field of cosmetics, in particular to a vegetable oil composition containing sweet wormwood oil and safflower seed oil and application thereof in promoting cell proliferation and activity.

Background

Skin barriers broadly include physical barriers, nerve barriers, immune barriers, and the like. Skin barrier is a narrow definition of a physical barrier, which we refer to at ordinary times. The physical barrier consists of sebum membrane, keratin, lipid, sandwich structure, brick wall structure, dermis mucopolysaccharide, etc., and has effects of resisting external harmful substances, irritants, sunlight, keeping moisture, and regulating antiinflammatory. The basic function of sebum is to moisten and keep moisture, and meanwhile, the weak acidity of sebum can play a certain antibacterial role. Sebum forms a "sebum film" on the skin surface, and is a natural moisturizing cream for skin. If sebum is lacking, the skin barrier function is weakened, the sebum contains triglyceride, free fatty acid, wax ester, squalene and other components, and the sebum film can be rebuilt on the skin surface by smearing the moisturizing cream, so that the external stimulus can be effectively isolated, and meanwhile, the moisture of the skin is prevented from being lost.

Depending on the source and chemical composition of the grease, the grease can be divided into: vegetable oil, animal oil, mineral oil and synthetic oil, the vegetable oil is close to the structure of human sebum membrane due to the skin-friendly texture, has good permeability, deeply permeates and nourishes the skin, and is favored by a plurality of skin care products. Vegetable oils and fats for cosmetic use are mainly derived from fruits, seeds and germs of plants, and also partially derived from leaves, barks, roots, petals, pistils and other parts of plants.

Plant bionic lipid technology (Phyto Bionic Sebum, abbreviated as PBS technology): natural vegetable oil is selected, natural lipid components in the skin barrier structure are simulated through scientific proportion, unsaturated fatty acids such as linolenic acid, linoleic acid and the like which are necessary for supplementing the skin cuticle are supplemented, and lipid components which are lack of skin are supplemented, so that the skin barrier is quickly repaired, and the skin is in a state of being healthy, moist and glossy.

The sweet wormwood oil is extracted from sweet wormwood, and is light yellow to brown in color, and is obtained by extracting sweet wormwood by supercritical extraction. Sweet wormwood crude oil mainly contains: arteannuic acid, alpha-linolenic acid, deoxyarteannuin, palmitic acid, etc., has effects in inhibiting inflammation such as IL-1a; increasing skin moisture related gene expression such as FLG, AQP3, LOR, etc.

Safflower seed oil contains 6% saturated fatty acids, 21% oleic acid, 73% linoleic acid (by weight). The main component of the food oil is linoleic acid, which is the oil with the highest linoleic acid in the food oil, so the food oil has high nutritive value and is of great interest in medicine, food, cosmetics and the like. For example, the literature, bole, research progress on safflower seed oil, world LATEST MEDICINE Information (ElectronicVersion) [ J ],2021, 21 (27): 146-148, it is pointed out that safflower seed oil has an antioxidant effect, an ability to effectively scavenge free radicals and reduce free radicals caused by chemical factors, and an anti-aging effect. The literature Jiang Li shows that the safflower seed oil can effectively prevent atherosclerosis, reduce the content of blood fat and serum cholesterol, soften blood vessels and indirectly recover various drug effect functions such as nervous system and the like after long-term consumption of the safflower seed oil by analyzing the efficacy and application prospect of the safflower seed oil and agricultural product processing [ J ],2017, 6:56-58.

Cell proliferation is a key mechanism for repair of skin lesions. The change of the proliferation activity of cells is detected based on keratinocytes, and the change is generally used for evaluating the effects of rejuvenating skin and repairing damage of an object to be tested.

With the research and development of biological cytology, there are many methods for researching cell viability in recent years, and special reagents are mainly used for measuring the metabolic viability of cells, including alamarblue, MTT and other tetrazolium salts. Alamar blue is a redox indicator that produces a change in brightness and a fluorescent signal based on cellular metabolic activity. The method is simple to operate and has no toxic effect, and is commonly used for detecting cell proliferation and cell viability in recent years. The MTT method mainly reflects the energy generation of cells, and utilizes the principle that mitochondrial dehydrogenase can decompose yellow MTT into blue-violet formazan energy in the growth and proliferation process of living cells to detect the change of the number and the activity of the cells, so that the MTT method is a simple and accurate common method for detecting the proliferation and the activity of the cells.

According to the invention, the vegetable oil composition of the sweet wormwood oil and the safflower seed oil is adopted for the first time, the proliferation and activity effects of the vegetable oil composition on skin epidermal cells are examined, and the unexpected finding that the effect of promoting cell proliferation and activity of the vegetable oil composition is obviously better than the effect of independently using the sweet wormwood oil and the safflower seed oil.

Disclosure of Invention

In one aspect, the present invention provides a vegetable oil composition for promoting cell proliferation and viability comprising sweet wormwood oil and safflower seed oil, wherein the volume ratio of sweet wormwood oil to safflower seed oil is equal to or less than 1:100, preferably the volume ratio is 1:100 to 1:800.

In a preferred embodiment, the sweet wormwood oil is obtained by extracting sweet wormwood by a supercritical extraction method.

In a preferred embodiment, the safflower seed oil comprises 50 to 90 wt% linoleic acid. More preferably, the safflower seed oil comprises 6 wt.% saturated fatty acids, 21 wt.% oleic acid, and 73 wt.% linoleic acid.

In another aspect, the present invention also relates to the use of a vegetable oil composition comprising sweet wormwood oil and safflower seed oil, wherein the volume ratio of sweet wormwood oil to safflower seed oil is equal to or less than 1:100, for the preparation of a medicament or a skin external preparation having an effect of promoting cell proliferation and viability.

In a preferred embodiment, the vegetable oil composition is contained in an amount of 0.001 to 20% by weight, preferably 0.01 to 5% by weight, in the pharmaceutical or skin external preparation.

In a preferred embodiment, the skin external agent is selected from the group consisting of: face cream, milky lotion, jelly, pack, face toilet, pack, aerosol cleansing foam, spray, body wash, or facial cleanser.

Brief description of the drawings

Figure 1 shows mass spectrum data of sweet wormwood oil. The sweet wormwood oil comprises 10-60% of sweet wormwood acid, 1-20% of alpha-linolenic acid and 1-20% of deoxyartemisinin by weight.

FIG. 2 shows the results of the plant oil compositions of comparative example 1 and examples 1-9 promoting cell viability (cell viability).

Detailed Description

The invention adopts the vegetable oil composition of the sweet wormwood oil and the safflower seed oil for the first time, and examines the proliferation and activity of the vegetable oil composition on skin epidermal cells. In addition, the vegetable oil composition containing sweet wormwood oil and safflower seed oil can be used as an efficacy combination additive to be applied to external skin preparations for promoting proliferation and activity of cells.

In order to provide a more concise description, some quantitative representations presented herein are not modified by the term "about". It will be understood that each quantity given herein is intended to refer to an actual given value, whether or not the term "about" is explicitly used, and is also intended to refer to approximations of such given values, including approximations of such given values resulting from experimental and/or measurement conditions, as reasonably deduced by one of ordinary skill in the art.

To provide a more concise description, some quantitative expressions herein are recited as a range from about X to about Y. It should be understood that when a range is recited, the range is not limited to the recited upper and lower limits, but rather, includes the entire range of about X to about Y amounts or any amount therebetween.

Sweet wormwood oil

In an embodiment of the present invention, sweet wormwood oil purchased from Shanghai household Biotechnology Co., ltd.

In a preferred embodiment, the sweet wormwood oil is extracted by supercritical fluid extraction. In some embodiments, the extraction process of sweet wormwood oil comprises the following steps: (a) Extracting herba Artemisiae Annuae with supercritical fluid to obtain crude extract of oleum Artemisiae Annuae; (b) re-dissolving the crude sweet wormwood oil extract and centrifuging; (c) And taking supernatant, decoloring, filtering and concentrating to obtain sweet wormwood oil.

In some embodiments, step (a) employs sweet wormwood having a particle size of 10-30 mesh. In a specific embodiment, step (a) employs sweet wormwood having a particle size of 20 mesh. In some embodiments, step (a) is performed at a pressure of 10 to 40 MPa. In a preferred embodiment, step (a) is carried out at a pressure of 20 to 40MPa or 30 to 40 MPa. In some embodiments, step (a) is performed at a temperature of 40-60 ℃. In a specific embodiment, step (a) is performed at a temperature of 50 ℃. In some embodiments, step (b) employs ethanol reconstitution, preferably 95% ethanol reconstitution. In some embodiments, the centrifugation conditions of step (b) are 3500rpm. In some embodiments, step (c) is decolorized with activated carbon powder. In a specific embodiment, supercritical extraction is performed using a south-Tong intelligent supercritical device RZSCF, 230-50-10L.

Fig. 1 shows the results of the composition analysis of sweet wormwood oil. As shown in FIG. 1, the sweet wormwood oil component preferably comprises 10-60% of sweet wormwood acid, 1-20% of alpha-linolenic acid and 1-20% of deoxyarteannuin by relative mass.

In some embodiments, the sweet wormwood oil is used at a concentration of at least 0.0001% by volume. In some embodiments, the sweet wormwood oil is used at a concentration of at least 0.000125% by volume.

Safflower seed oil

The main component of the safflower seed oil is linoleic acid, which is the highest linoleic acid in the food oil, so the safflower seed oil has high nutritive value and is of great interest in medicine, food, cosmetics and the like. Safflower seed oil contains 50-90 wt% linoleic acid. In a specific embodiment, the safflower seed oil comprises 6% saturated fatty acids, 21% oleic acid, 73% linoleic acid by weight.

In an embodiment of the present invention, safflower seed oil from the company natural vegetable oil, north british (Northstar Lipids UK ltd.) was used.

In some embodiments, safflower seed oil is used at a concentration of at least 0.01% by volume. In some embodiments, safflower seed oil is used at a concentration of at least 0.0125% by volume.

In some embodiments, safflower seed oil is used at a concentration of 0.0125% to 0.1% by volume.

In some embodiments, the volume ratio of sweet wormwood oil to safflower seed oil is equal to or less than 1:100. In some embodiments, the volume ratio of sweet wormwood oil to safflower seed oil is 1:100 to 1:800.

External preparation for skin containing vegetable oil composition

The vegetable oil composition of the present invention can be applied as an efficacy additive to skin external preparations for promoting cell proliferation and viability, thereby improving skin conditions.

In some embodiments, the vegetable oil composition is present in the skin external agent in an amount of 0.001 to 20% by weight, preferably 0.01 to 5% by weight.

In some embodiments, the skin external agent is selected from the group consisting of: face creams, lotions, gels, lotions, essences, face masks, eye creams, aerosols (cleansing bubbles), sprays, body washes and facial washes. Different amounts are added according to the different types of formulations.

The external skin preparation is a general concept of all ingredients commonly used outside the skin, and may be, for example, a cosmetic composition. The cosmetic composition may be basic cosmetic, facial makeup cosmetic, body cosmetic, hair care cosmetic, etc., and its dosage form is not particularly limited and may be reasonably selected according to different purposes. The cosmetic composition also contains various cosmetically acceptable medium or matrix excipients depending on dosage form and purpose.

The external preparation for skin contains a dermatologically acceptable carrier or vehicle (e.g., a lotion, a cream, an ointment, a cleanser, etc.). The person of ordinary skill in the art will be able to select a carrier that will dissolve or disperse these components at the concentrations described above, in accordance with common general knowledge in the art. When a carrier is used, the carrier should not cause inactivation of the vegetable oil composition and should not cause any adverse effect on the skin upon use.

The person of ordinary skill in the art will be able to select suitable carriers, including, for example, water, alcohols, oils, etc., based on common general knowledge and their ability to dissolve or disperse in the active ingredient at a concentration most suitable for treatment.

The skin external preparation of the present invention may be in the form of a topical application product which can be externally applied to the skin and can be prepared by those ordinary techniques well known in the art. The carrier may take a variety of practical forms, such as creams, dressings, gels, lotions, ointments or liquids, including compositions for application and cleansing, and incorporating them into a carrier material such as a dry or wet spread, hydrogel matrix, or adhesive (or non-adhesive) patch by methods well known in the art. Preferably, the carrier is a gel or a moisturizing lotion, or an application in dry or wet form.

Typical carriers include emulsions comprising water and/or an alcohol and an emollient, wherein the emollient is, for example, a hydrocarbon oil and wax, silicone oil, hyaluronic acid, a vegetable, animal or marine fat or oil, a glyceride derivative, a fatty acid, or fatty acid ester or alcohol ether, lanolin and its derivatives, a polyol or ester, a wax ester, a sterol, a phospholipid, and the like, and typically also an emulsifier (nonionic, cationic or anionic), although some emollients themselves have emulsifying properties. In addition, these same components may be formulated into creams, gels, or solid sticks using different proportions of their components and/or by incorporating thickeners such as gums or other forms of hydrocolloids.

The skin external agent of the present invention may contain additional components commonly found in skin care compositions, such as emollients, skin conditioning agents, emulsifiers, preservatives, antioxidants, fragrances, chelating agents, and the like, as long as they are physically and chemically compatible with the other components of the skin external agent and do not affect the effects of the vegetable oil composition of the present invention.

In some embodiments of the skin external preparation of the present invention, one or more preservatives may be used. Suitable preservatives include p-hydroxyacetophenone, alkyl C1-C4 p-hydroxybenzoates and phenoxyethanol. The preservative is used in an amount of about 0.5 to about 2 wt%, preferably about 0.5 to 1 wt%, based on the total weight of the composition.

In one example of the skin external agent of the present invention, one or more antioxidants may be used. Suitable antioxidants include Butylated Hydroxytoluene (BHT), ascorbyl palmitate (BHA), butylated hydroxyanisole, phenyl-alpha-naphthylamine, hydroquinone, propyl gallate, nordihydroguaiaretic acid, vitamin E or derivatives of vitamin E, vitamin C and its derivatives, calcium pantothenate, green tea extracts and mixed polyphenols, and mixtures of the foregoing. The antioxidants are used in an amount ranging from about 0.02 to 0.5 weight percent, more preferably from about 0.002 to 0.1 weight percent, based on the total weight of the composition.

In one example of the skin external agent of the present invention, one or more emollients may be used which act as lubricants to reduce flaking and improve the appearance of the skin by their ability to remain on the skin surface or in the stratum corneum. Typical emollients include fatty esters, fatty alcohols, mineral oils, polyether siloxane copolymers, and the like. Examples of suitable emollients include, without limitation, polypropylene glycol ("PPG") -15 stearyl ether, PPG-10 cetyl ether, steareth-10, oleth-8, PPG-4 lauryl ether, vitamin E acetate, lanolin, cetyl alcohol, cetostearyl alcohol ethyl hexanoate, cetostearyl alcohol, glyceryl stearate, octyl hydroxystearate, dimethylpolysiloxane, and combinations thereof. Cetyl alcohol, cetostearyl alcohol ethyl hexanoate, cetostearyl alcohol, glycerol stearate, and combinations thereof are preferred. When used, the emollient is in an amount ranging from about 0.1 to about 30 weight percent, preferably from about 1 to about 30 weight percent, based on the total weight of the composition.

In one example of the skin external agent of the present invention, one or more moisturizers may be used. Humectants, also known as humectants, help to enhance the effectiveness of emollients, reduce flaking, stimulate removal of constituent scales and enhance skin feel. Polyols may be used as humectants including, but not limited to, glycerin, polyalkylene glycols, alkylene polyols and derivatives thereof, including butylene glycol, propylene glycol, dipropylene glycol, polyglycerol, polyethylene glycol and derivatives thereof, sorbitol, hydroxypropyl sorbitol, hexylene glycol, 1, 3-dibutylene glycol, 1,2, 6-hexanetriol, ethoxylated glycerin, propoxylated glycerin and combinations thereof. When used, the humectant is present in an amount of about 0.1 to about 20 weight percent, preferably about 1 to about 15 weight percent, based on the total weight of the composition.

In one example of the skin external agent of the present invention, one or more emulsifying agents may be used. The emulsifier may be used in an effective stabilizing amount. Preferably, the emulsifier is used in an amount of about 1.0 to about 10.0 wt%, more preferably about 3.0 to about 6.0 wt%, based on the total weight of the composition. Any emulsifier that is compatible with the components of the composition may be used. Suitable emulsifiers include stearic acid, cetyl alcohol, glyceryl stearate, lecithin, stearyl alcohol, steareth-2, steareth-20, acrylic/C10-30 alkanol acrylate cross-linked polymers, and combinations thereof.

In one example of the skin external agent of the present invention, one or more pH adjusting agents may be used. The pH adjuster useful in the skin external preparation of the present invention includes tromethamine. When used, the pH adjustor is used in an amount of about 0.1 to about 2 weight percent, preferably about 0.1 to about 1 weight percent, based on the total weight of the composition.

In one embodiment of the present invention, the skin external preparation comprises acrylic/C10-30 alkanol acrylate cross-linked polymer, glycerol, p-hydroxyacetophenone, glycerol stearate and lecithin, cetyl/stearyl alcohol, cetostearyl alcohol ethyl hexanoate, tromethamine or combinations thereof.

In some embodiments of the present invention, the vegetable oil composition is used in an amount of 0.001% -20% (w/w), preferably 0.01% -20% (w/w), more preferably 0.01% -10% (w/w), and most preferably 0.1% -5% (w/w) of the skin external preparation.

Examples

The invention will be further illustrated by the following examples. It should be noted that the scope of the present invention is not limited to these embodiments, but is intended to illustrate the technical solution of the present invention and not to limit the scope of the present invention. The experimental procedures, which do not address the specific conditions in the examples below, are generally performed under conventional conditions or under conditions recommended by the manufacturer. The percentages of sweet wormwood oil and safflower seed oil in the following comparative examples and examples are calculated by volume.

1. Experimental materials

Safflower (CARTHAMUS TINCTORIUS) seed oil, trade name safflower seed oil, purchased from Northstar Lipids UK ltd;

sweet wormwood oil is purchased from Shanghai household Biotechnology Co.

2. Instrument and reagent

Instrument: bench top centrifuges, CO ₂ incubator HERAcell i, microplate reader Multiskan Mk3 (Thermo), biosafety cabinet 1300 services A2, and the like.

Reagent: MTT assay kit (Sigma), DMEM high-glucose medium (Gibco 11995065), FBS, PBS (Soxhibao), dimethyl sulfoxide (DMSO, sigma).

3. Reagent configuration:

DMEM complete medium: adding FBS into DMEM high sugar culture medium to make its content 10%, and storing at 4deg.C.

Comparative example 1 cell culture medium:

According to the calculation, the DMSO addition amount in each example is not more than 0.1%, and the culture medium in comparative example 1 is selected from DMEM complete culture medium with DMSO content of 0.1% because the culture medium has no cytotoxicity in combination with previous data that the DMSO addition amount is less than 0.1%.

Comparative example 1 cell culture medium: 1ml of comparative example 1 cell culture medium = 1 μl DMSO +999 μ lDMEM complete medium, a culture medium of comparative example 1 was obtained with DMSO content of 0.01%.

Example 1 cell culture medium:

Preparing mother solution: sweet wormwood oil and DMSO are prepared according to a volume ratio of 1:499.

Example 1 cell culture medium: 1ml of example 1 cell culture medium = 0.625 μl of mother liquor +999.375 μl of DMEM complete medium, resulting in the culture medium of example 1 having an oil content of 0.000125% in artemisia annua.

Example 2 cell culture medium:

preparing mother solution: safflower seed oil and DMSO are prepared according to a volume ratio of 1:1.

Example 2 cell culture medium: 1ml of cell culture medium of example 2 = 0.25 μl of mother liquor +999.75 μ lDMEM complete medium, resulting in the culture medium of example 2 with safflower seed oil content of 0.0125%.

Example 3 cell culture medium:

Example 3 cell culture medium: 1ml of cell culture medium of example 3 = 0.5 μl of mother liquor +999.5 μ lDMEM complete medium, resulting in the culture medium of example 3 having a safflower seed oil content of 0.025%.

Example 4 cell culture medium:

Example 4 cell culture medium: 1ml of cell culture medium of example 4 = 1 μl of mother liquor +999 μl of DMEM complete medium, resulting in the culture medium of example 4 with safflower seed oil content of 0.05%.

Example 5 cell culture medium:

Example 5 cell culture medium: 1ml of cell culture medium of example 5 = 2 μl mother liquor +998 μl DMEM complete medium, resulting in the culture medium of example 5 with safflower seed oil content of 0.1%.

Example 6 cell culture medium:

preparing mother solution: sweet wormwood oil: safflower seed oil: DMSO was formulated at a volume ratio of 1:100:99.

Example 6 cell culture medium: 1ml of cell culture medium of example 1 = 0.25 μl of mother liquor +999.75 μ lDMEM complete medium, resulting in the medium of example 6 having an sweet wormwood oil content of 0.000125% and a safflower seed oil content of 0.125%.

Example 7 cell culture medium:

preparing mother solution: sweet wormwood oil: safflower seed oil: DMSO was formulated at a volume ratio of 1:200:199.

Example 7 cell culture medium: 1ml of cell culture medium of example 1 = 0.5 μl of mother liquor +999.5 μ lDMEM complete medium, resulting in the medium of example 7 having an sweet wormwood oil content of 0.000125% and a safflower seed oil content of 0.025%.

Example 8 cell culture medium:

Preparing mother solution: sweet wormwood oil: safflower seed oil: DMSO was configured at a volume ratio of 0.5:200:199.5.

Example 8 cell culture medium: 1ml of example 1 cell culture medium = 1 μl of mother liquor +999 μl of DMEM complete medium, resulting in the culture medium of example 8 having an sweet wormwood oil content of 0.000125% and a safflower seed oil content of 0.05%.

Example 9 cell culture medium:

Preparing mother solution: sweet wormwood oil: safflower seed oil: DMSO was formulated at a volume ratio of 0.5:400:399.5.

Example 9 cell culture medium: 1ml of cell culture medium of example 1 = 2 μl mother liquor +998 μl DMEM complete medium, resulting in the culture medium of example 9 having an sweet wormwood oil content of 0.000125% and a safflower seed oil content of 0.1%.

Table 1 summarizes the amounts and proportions of sweet wormwood oil and safflower seed oil per ml of medium in the examples.

TABLE 1

Test example 1: comparative example 1 and examples 1, 2, 6 keratinocyte viability test

The invention adopts a method for testing the cell viability of human immortalized keratinocytes Hacat, and adds a proper amount of samples prepared in the examples into keratinocytes and incubates for 24 hours. After 24 hours, the samples were washed away and tested for cell viability using the MTT method. The ability of the samples of the examples to promote cell proliferation and cell viability can be evaluated based on the increase in viability.

Hacat cell seed plate

The medium in the T175 flask was discarded, washed once with PBS, digested with 0.25% EDTA trypsin for 1min, then fresh medium was added to stop the digestion, the cell suspension was transferred to a 15mL centrifuge tube, centrifuged at 800rpm for 5min, and the supernatant was discarded. Cell counts were performed after resuspension with DMEM complete medium. The cell suspension was diluted to a density of 8000 cells/100. Mu.l and inoculated in 96-well plates at 100. Mu.l/well, and after the completion of the inoculation, cultured in a 5% CO2 incubator at 37℃for 24 hours.

Cell processing and sample addition

Taking out the 96-well plate, discarding the supernatant, adding the culture medium corresponding to comparative example 1 and examples 2, 3 and 6, 100 mu L/well, and culturing in a 5% CO2 incubator at 37 ℃ for 24 hours;

Cell proliferation and viability assays

Preparing MTT mother liquor: 25mg of MTT was dissolved in 5ml of MTT solvent to prepare a 5mg/ml MTT solution.

MTT solution configuration: the preparation was carried out in a ratio of 10. Mu. lMTT solution to 90. Mu.l of DMEM complete medium.

Sample adding: the original medium was removed, 100. Mu.l of the MTT solution described above was added to each well, and incubation was continued for 4 hours in the cell incubator.

Dissolving: the MTT solution was carefully removed, 100. Mu.l DMSO was added to each well and mixed well and incubation was continued in the cell incubator. Until the crystalline dark purple product formazan was found to be totally dissolved by observation under a common light microscope.

Absorbance measurement: the absorbance was measured at 570nm by a microplate reader.

Result judgment

Comparative example 1 is an unslotted group with relative viability of 100%; the examples containing sweet wormwood oil and safflower seed oil were compared with comparative example 1 and examples containing equal amounts of sweet wormwood oil and safflower seed oil alone. If the cell proliferation rate and the cell activity are obviously higher than those of other groups, the composition is considered to have better effects of promoting the cell proliferation and the cell activity, and has better effects of reviving skin and repairing injury.

Experimental results

As shown in Table 2, compared with the sweet wormwood oil and the safflower seed oil which are singly used in the same concentration, the sweet wormwood oil and the safflower seed oil are compounded according to the volume ratio of 1:100, and the effect of obviously enhancing the cell proliferation and the activity is achieved.

TABLE 2

The above p-value calculations are all compared with example 6.

Test example 2: comparative example 1 and examples 1, 3, 7 keratinocyte viability test

The test was performed using the same experimental procedure as in test example 1.

Experimental results:

As shown in Table 3, compared with the sweet wormwood oil and the safflower seed oil which are singly used in the same concentration, the sweet wormwood oil and the safflower seed oil are compounded according to the volume ratio of 1:200, and the effect of obviously enhancing the cell proliferation and the activity is achieved.

TABLE 3 Table 3

The above p-value calculations are all compared with example 7.

Test example 3: comparative example 1 and examples 1, 4, 8 keratinocyte viability test

Experimental results:

as shown in Table 4, compared with the sweet wormwood oil and the safflower seed oil which are singly used in the same concentration, the sweet wormwood oil and the safflower seed oil are compounded according to the volume ratio of 1:400, and the effect of obviously enhancing the cell proliferation and the activity is achieved.

TABLE 4 Table 4

The above p-value calculations are all compared with example 8.

Test example 4: comparative example 1 and examples 1, 5, 9 keratinocyte viability test

Experimental results:

As shown in Table 5, compared with the sweet wormwood oil and the safflower seed oil which are singly used in the same concentration, the sweet wormwood oil and the safflower seed oil are compounded according to the volume ratio of 1:800, and the effect of obviously enhancing the cell proliferation and the activity is achieved.

TABLE 5

The above p-value calculations are all compared with example 9.

The vegetable oil composition can be used as an efficacy additive and applied to skin external preparations, and the skin external preparations are preferably cosmetic compositions, including but not limited to preparation of products in forms of face cream, emulsion, gel, toning lotion, essence, facial mask, eye cream, aerosol (cleaning foam), spray, bath lotion, facial cleanser and the like. For example, the vegetable oil compositions of examples 6,7, 8 and 9 were used in an amount of 0.0001% to 20% (w/w) by weight in the skin external preparation. Preferably 0.001% to 10% (w/w). More preferably 0.001% to 5% (w/w). Most preferably 0.01% -5% (w/w) by weight.

The following are specific examples of application of the vegetable oil composition of the examples in skin external preparations, and formulations and preparation methods of these dosage forms. Specific applications are as follows:

Application example 1: preparation of face cream

Application example 2: preparation of the emulsion

Application example 3: preparation of eye cream

Application example 4: preparation of facial mask

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Application example 5: preparation of essence

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Claims

1. A vegetable oil composition that promotes cell proliferation and viability, the vegetable oil composition comprising sweet wormwood oil and safflower seed oil, wherein the volume ratio of sweet wormwood oil to safflower seed oil is equal to or less than 1:100.

2. The vegetable oil composition of claim 1, wherein the volume ratio of sweet wormwood oil to safflower seed oil is 1:100 to 1:800.

3. The vegetable oil composition according to claim 1, wherein the sweet wormwood oil is obtained by extracting sweet wormwood herb by a supercritical extraction method.

4. The vegetable oil composition of claim 1, wherein the safflower seed oil comprises 50-90 wt.% linoleic acid.

5. The vegetable oil composition of claim 1, wherein the safflower seed oil comprises 6 wt.% saturated fatty acids, 21 wt.% oleic acid, and 73 wt.% linoleic acid.

6. Use of a vegetable oil composition comprising sweet wormwood oil and safflower seed oil in a volume ratio of sweet wormwood oil to safflower seed oil of 1:100 or less for the preparation of a medicament or a skin external preparation having an effect of promoting cell proliferation and viability.

7. The use according to claim 6, wherein the sweet wormwood oil is obtained by extracting sweet wormwood by supercritical extraction.

8. The use according to claim 6, wherein the safflower seed oil comprises 50-90% by weight linoleic acid.

9. The use according to claim 6, wherein the safflower seed oil comprises 6 wt.% saturated fatty acids, 21 wt.% oleic acid and 73 wt.% linoleic acid.

10. The use as claimed in claim 6, wherein the vegetable oil composition is contained in an amount of 0.001 to 20% by weight in the pharmaceutical or external skin preparation.

11. The use as claimed in claim 10, wherein the vegetable oil composition is contained in an amount of 0.01 to 5% by weight in the pharmaceutical or dermatological external preparation.

12. The use according to claim 6, wherein the external skin preparation is selected from the group consisting of: face cream, milky lotion, jelly, pack, face toilet, pack, aerosol cleansing foam, spray, body wash, or facial cleanser.