Plant oil compositions that promote cell proliferation and viability

Abstract

The present application provides a plant oil composition for promoting cell proliferation and viability, which comprises artemisia oil and safflower seed oil, wherein the volume ratio of artemisia oil to safflower seed oil is equal to or less than 1:100. The artemisia oil of the present application is extracted from Artemisia annua by supercritical extraction method. The safflower seed oil of the present application comprises 50-90 wt% of linoleic acid. The present application also relates to the use of the plant oil composition in the preparation of a drug or a skin external preparation for promoting cell proliferation and viability.

Description

Technical Field

[0001] This invention relates to the field of cosmetics, and more specifically to plant oil compositions containing artemisia annua oil and safflower seed oil, and their application in promoting cell proliferation and vitality. Background Technology

[0002] The skin barrier, broadly speaking, includes physical, neural, and immune barriers. In a narrower sense, it primarily refers to the physical barrier, which is what we usually refer to when we talk about the skin barrier. The physical barrier is composed of the sebum film, keratin in the stratum corneum, lipids, a "sandwich" structure, a brick-and-mortar structure, dermal mucopolysaccharides, and other components. It defends against harmful external substances, irritants, and sunlight, while also providing moisturizing and anti-inflammatory benefits. The basic function of sebum is moisturizing, and its slightly acidic nature also contributes to its antibacterial properties. Sebum forms the "sebum film" on the skin's surface, acting as the skin's natural moisturizer. A lack of sebum weakens the skin barrier function. Sebum contains triglycerides, free fatty acids, wax esters, squalene, and other components. Applying moisturizer can rebuild the sebum film on the skin's surface, effectively isolating external irritants and preventing moisture loss.

[0003] Based on their source and chemical composition, oils can be mainly classified into: plant oils, animal oils, mineral oils, and synthetic oils. Plant oils, due to their skin-friendly texture, similar structure to human sebum, and good permeability, deeply penetrate and nourish the skin, making them a favorite in many skincare products. Plant oils used in cosmetics mainly come from plant fruits, seeds, and germs, and some also come from other parts of the plant such as leaves, bark, roots, petals, and stamens.

[0004] Phyto Bionic Sebum (PBS) technology: It selects natural plant oils and uses scientific proportions to simulate the natural lipid components in the skin barrier structure, replenishing the skin's stratum corneum with essential unsaturated fatty acids, such as linolenic acid and linoleic acid, and replenishing the skin's missing lipid components. It is compatible with sebum, thereby quickly repairing the skin barrier and restoring the skin to a healthy, moisturized, and radiant state.

[0005] Artemisia annua oil is extracted from Artemisia annua, and its color ranges from pale yellow to brown. It is obtained by supercritical fluid extraction. Artemisia annua oil mainly contains artemisinic acid, α-linolenic acid, deoxyartemisinin, palmitic acid, etc., and has the effects of inhibiting inflammation such as IL-1a; increasing the expression of skin moisturizing-related genes, such as FLG, AQP3, and LOR.

[0006] Safflower seed oil contains 6% saturated fatty acids, 21% oleic acid, and 73% linoleic acid (by weight). Since its main component is linoleic acid, it has the highest linoleic acid content among edible oils, making it highly nutritious and attracting significant attention in medicine, food, and cosmetics. For example, the literature "Research Progress of Safflower Seed Oil," World Latest Medicine Information (Electronic Version) [J], 2021, 21(27): 146-148, points out that safflower seed oil has antioxidant properties, effectively scavenging free radicals and reducing free radicals caused by chemical factors, while also possessing anti-aging effects. The literature "Efficacy and Application Prospect Analysis of Safflower Seed Oil," Agricultural Product Processing [J], 2017, 6: 56-58, indicates that long-term consumption of safflower seed oil can effectively prevent atherosclerosis, reduce blood lipids and serum cholesterol levels, soften blood vessels, and indirectly restore the nervous system, among other pharmacological functions.

[0007] Cell proliferation is a key mechanism for skin damage repair. Detecting changes in the proliferative activity of keratinocytes is commonly used to evaluate the skin-revitalizing and damage-repairing effects of test products.

[0008] With the advancement of cell biology research, numerous methods for studying cell viability have emerged in recent years. These primarily utilize specialized reagents to measure cellular metabolic activity, including alamar blue, MTT, and other tetrazolium salts. Alamar blue is a redox indicator that produces brightness changes and fluorescence signals based on cellular metabolic activity. It is simple to use, non-toxic, and has become increasingly common for detecting cell proliferation and viability. The MTT assay primarily reflects cellular energy production. It utilizes the principle that mitochondrial dehydrogenases break down yellow MTT into blue-purple formazan during cell growth and proliferation to detect changes in cell number and viability. This is a simple and accurate method for detecting cell proliferation and viability.

[0009] This invention is the first to use a plant oil composition of artemisia annua oil and safflower seed oil to investigate its effects on the proliferation and vitality of skin epidermal cells. Unexpectedly, it was found that the plant oil composition has a significantly better effect on promoting cell proliferation and vitality than the two oils used alone. Summary of the Invention

[0010] On one hand, the present invention provides a plant oil composition that promotes cell proliferation and vitality, the plant oil composition comprising artemisia annua oil and safflower seed oil, wherein the volume ratio of artemisia annua oil to safflower seed oil is equal to or less than 1:100, preferably the volume ratio is 1:100 to 1:800.

[0011] In a preferred embodiment, the artemisia oil is obtained by supercritical extraction of Artemisia annua.

[0012] In a preferred embodiment, the safflower seed oil contains 50-90% by weight of linoleic acid. More preferably, the safflower seed oil contains 6% by weight of saturated fatty acids, 21% by weight of oleic acid, and 73% by weight of linoleic acid.

[0013] On the other hand, the present invention also relates to the use of a plant oil composition in the preparation of a pharmaceutical or topical skin agent that promotes cell proliferation and vitality, said plant oil composition comprising artemisia annua oil and safflower seed oil, wherein the volume ratio of artemisia annua oil to safflower seed oil is equal to or less than 1:100.

[0014] In a preferred embodiment, the content of the vegetable oil composition in the pharmaceutical or topical skin preparation is 0.001-20% by weight, preferably 0.01-5% by weight.

[0015] In a preferred embodiment, the topical skin agent is selected from: face cream, lotion, gel, toner, serum, face mask, eye cream, aerosol cleansing foam, spray, shower gel, or facial cleanser. Brief description of the attached figures

[0016] Figure 1 Mass spectrometry data for artemisia annua oil are shown. By weight, artemisia annua oil contains 10-60% artemisinic acid, 1-20% α-linolenic acid, and 1-20% deoxyartemisinin.

[0017] Figure 2 The results of the vegetable oil compositions of Comparative Example 1 and Examples 1-9 promoting cell viability are shown. Detailed Implementation

[0018] This invention is the first to utilize a plant oil composition of artemisia annua oil and safflower seed oil to investigate its effects on the proliferation and vitality of skin epidermal cells. Furthermore, this plant oil composition containing artemisia annua oil and safflower seed oil can be used as a functional additive in topical skin preparations to promote cell proliferation and vitality.

[0019] To provide a more concise description, some of the quantitative expressions given herein are not modified by the term "approximately". It should be understood that, whether or not the term "approximately" is explicitly used, each quantity given herein is intended to refer to an actual given value, and also to an approximation of such given values ​​that can be reasonably inferred by one of ordinary skill in the art, including approximations of such given values ​​caused by experimental and / or measurement conditions.

[0020] To provide a more concise description, some quantities in this document are described as a range from approximately X to approximately Y. It should be understood that when describing a range, the range is not limited to the upper and lower limits stated, but should include the entire range from approximately X to approximately Y, or any quantities in between.

[0021] Artemisia oil

[0022] Artemisia annua oil is extracted from Artemisia annua, and its color ranges from pale yellow to brown. It is obtained by supercritical fluid extraction. Artemisia annua oil mainly contains artemisinic acid, α-linolenic acid, deoxyartemisinin, palmitic acid, etc., and has the effects of inhibiting inflammation such as IL-1a; increasing the expression of skin moisturizing-related genes, such as FLG, AQP3, and LOR.

[0023] In an embodiment of the present invention, artemisia oil purchased from Shanghai Jahwa Biotechnology Co., Ltd. was used.

[0024] In a preferred embodiment, supercritical fluid extraction is used to extract artemisia oil. In some embodiments, the extraction process of artemisia oil includes the following steps: (a) taking artemisia and subjecting it to supercritical fluid extraction to obtain crude artemisia oil extract; (b) redissolving the obtained crude artemisia oil extract and centrifuging; (c) taking the supernatant, decolorizing, filtering, and concentrating to obtain artemisia oil.

[0025] In some embodiments, step (a) uses Artemisia annua with a particle size of 10-30 mesh. In one specific embodiment, step (a) uses Artemisia annua with a particle size of 20 mesh. In some embodiments, step (a) is carried out at a pressure of 10-40 MPa. In a preferred embodiment, step (a) is carried out at a pressure of 20-40 MPa or 30-40 MPa. In some embodiments, step (a) is carried out at a temperature of 40-60°C. In one specific embodiment, step (a) is carried out at a temperature of 50°C. In some embodiments, step (b) involves redissolving the sample in ethanol, preferably 95% ethanol. In some embodiments, step (b) is centrifuged at 3500 rpm. In some embodiments, step (c) involves decolorization using activated carbon powder. In a specific embodiment, supercritical extraction is performed using a supercritical fluid analyzer RZSCF230-50-10L from Nantong Ruizhi.

[0026] Figure 1 The results of the component analysis of artemisia annua oil are shown. For example... Figure 1 As shown, the preferred components of artemisia oil include artemisinin, α-linolenic acid, and deoxyartemisinin, with a relative mass content of 10-60%.

[0027] In some embodiments, the concentration of artemisia oil used is at least 0.0001% by volume. In some embodiments, the concentration of artemisia oil used is at least 0.000125% by volume.

[0028] Safflower seed oil

[0029] Safflower seed oil's main component is linoleic acid, making it the oil with the highest linoleic acid content among edible oils. Therefore, it has high nutritional value and is attracting attention in medicine, food, and cosmetics. Safflower seed oil contains 50-90% by weight of linoleic acid. In one specific embodiment, safflower seed oil contains, by weight, 6% saturated fatty acids, 21% oleic acid, and 73% linoleic acid.

[0030] In an embodiment of the present invention, safflower seed oil purchased from NorthstarLipids UK Ltd. is used.

[0031] In some embodiments, the concentration of safflower seed oil used is at least 0.01% by volume. In some embodiments, the concentration of safflower seed oil used is at least 0.0125% by volume.

[0032] In some embodiments, the concentration of safflower seed oil used is 0.0125% to 0.1% by volume.

[0033] In some embodiments, the volume ratio of artemisia annua oil to safflower seed oil is equal to or less than 1:100. In some embodiments, the volume ratio of artemisia annua oil to safflower seed oil is from 1:100 to 1:800.

[0034] Topical skin preparations containing plant oil compositions

[0035] The plant oil composition of the present invention can be used as an efficacy additive in topical skin preparations to promote cell proliferation and vitality, thereby improving skin condition.

[0036] In some embodiments, the vegetable oil composition is present in the topical skin preparation at a concentration of 0.001-20% by weight, preferably 0.01-5% by weight.

[0037] In some embodiments, the topical skin agent is selected from: face creams, lotions, gels, toners, serums, masks, eye creams, aerosols (cleansing foams), sprays, shower gels, and facial cleansers. Different dosages are added depending on the type of formulation.

[0038] The term "topical skin agent" is a general term encompassing all ingredients typically used on the external surface of the skin, such as cosmetic compositions. These cosmetic compositions can include basic cosmetics, facial makeup cosmetics, body cosmetics, hair care cosmetics, etc., with no specific restrictions on their dosage forms; they can be rationally selected according to different purposes. Depending on the dosage form and purpose, these cosmetic compositions may also contain different cosmetically permissible media or matrix excipients.

[0039] Topical skin preparations contain dermatologically acceptable carriers or mediators (e.g., lotions, creams, ointments, cleansers, etc.). Those skilled in the art can select carriers capable of dissolving or dispersing these components at the concentrations described above, based on common knowledge in the art. When using a carrier, it should not cause inactivation of the vegetable oil composition and should not produce any adverse effects on the skin during application.

[0040] Those skilled in the art can select suitable carriers based on common knowledge and their ability to dissolve or disperse in the active component at the concentration most suitable for processing, such as water, alcohols, oils, etc.

[0041] The topical skin agents of the present invention can be in the form of topical application products that can be applied externally to the skin and can be prepared using common techniques known in the art. The carrier can have various practical forms, such as creams, dressings, gels, lotions, ointments, or liquids, including compositions that are applied and washed off, and materials incorporated into them using methods known in the art, such as dry or wet applicators, hydrogel matrices, or adhesive (or non-adhesive) patches. Preferably, the carrier is a gel or a moisture-enhancing lotion, or an applicator in dry or wet form.

[0042] Typical carriers include emulsions containing water and / or alcohols and emollients, where the emollients are, for example, oils and waxes of hydrocarbons, silicone oils, hyaluronic acid, fats or oils from plants, animals, or marine organisms, glyceryl ester derivatives, fatty acids, or fatty acid esters or alcohols or alcohol ethers, lanolin and its derivatives, polyols or esters, wax esters, sterols, phospholipids, etc., and generally include emulsifiers (nonionic, cationic, or anionic), although some emollients themselves have emulsifying properties. Additionally, these same components can be formulated into creams, gels, or solid bars by utilizing different proportions of their components and / or by incorporating thickeners such as gums or other forms of hydrophilic colloids.

[0043] The topical skin preparations of the present invention may include additional components commonly found in skin care compositions, such as emollients, skin conditioners, emulsifiers, preservatives, antioxidants, fragrances, chelating agents, etc., as long as they are physically and chemically compatible with other components in the topical skin preparations and do not affect the efficacy of the plant oil compositions of the present invention.

[0044] In some embodiments of the topical skin formulation of the present invention, one or more preservatives may be used. Suitable preservatives include p-hydroxyacetophenone, C1-C4 alkyl p-hydroxybenzoate, and phenoxyethanol. Based on the total weight of the composition, the amount of preservative used is from about 0.5 to about 2% by weight, preferably from about 0.5 to 1% by weight.

[0045] In one example of the topical skin preparation of the present invention, one or more antioxidants may be used. Suitable antioxidants include butylated hydroxytoluene (BHT), ascorbyl palmitate (BHA), butylated hydroxyanisole, phenyl-α-naphthylamine, hydroquinone, propyl gallate, nordihydroguaiac acid, vitamin E or a derivative of vitamin E, vitamin C and its derivatives, calcium pantothenate, green tea extract, and mixed polyphenols, as well as mixtures of the substances described above. The antioxidant used is approximately 0.02 to 0.5% by weight of the total weight of the composition, more preferably in the range of approximately 0.002 to 0.1% by weight.

[0046] In one example of the topical skin preparation of the present invention, one or more emollients may be used, which, by virtue of their ability to remain on the skin surface or in the stratum corneum, act as lubricants to reduce exfoliation and improve the appearance of the skin. Typical emollients include fatty esters, fatty alcohols, mineral oils, polyether silicone copolymers, and the like. Examples of suitable emollients, without limitation, include polypropylene glycol (“PPG”)-15 octadecyl ether, PPG-10 hexadecyl ether, Steareth-10, Oleth-8, PPG-4 dodecyl ether, vitamin E acetate, lanolin, cetyl alcohol, cetearyl alcohol ethylhexanoate, cetearyl alcohol, glyceryl stearate, octyl hydroxystearate, dimethyl polysiloxane, and combinations thereof. Cetyl alcohol, cetearyl alcohol ethylhexanoate, cetearyl alcohol, glyceryl stearate, and combinations thereof are preferred. When used, the emollient is applied in an amount ranging from about 0.1% to about 30% by weight, preferably from about 1% to about 30% by weight, based on the total weight of the composition.

[0047] In one example of the topical skin preparation of the present invention, one or more moisturizers may be used. Moisturizers, also known as humectants, help enhance the effectiveness of emollients, reduce exfoliation, stimulate the removal of scaly skin, and improve skin feel. Polyols may be used as moisturizers, including, but not limited to, glycerin, polyalkylene glycols, alkylene polyols and their derivatives, including butylene glycol, propylene glycol, dipropylene glycol, glycerol, polyethylene glycol and their derivatives, sorbitol, hydroxypropyl sorbitol, hexanediol, 1,3-dibutylene glycol, 1,2,6-hexanetriol, ethoxylated glycerin, propoxylated glycerin, and combinations thereof. When used, the amount of moisturizer is from about 0.1% to about 20% by weight, preferably from about 1% to about 15% by weight, based on the total weight of the composition.

[0048] In one example of the topical skin formulation of the present invention, one or more emulsifiers may be used. The emulsifier may be used within a range of effective stable amounts. Preferably, the emulsifier is used in an amount of about 1.0 to about 10.0% by weight, more preferably about 3.0 to about 6.0% by weight, based on the total weight of the composition. Any emulsifier compatible with the components in the composition may be used. Suitable emulsifiers include stearic acid, cetyl alcohol, glyceryl stearate, lecithin, octadecyl alcohol, Steareth-2, Steareth-20, acrylate / C10-30 alkanol acrylate crosspolymers, and combinations thereof.

[0049] In one example of the topical skin formulation of the present invention, one or more pH adjusters may be used. Beneficial pH adjusters in the topical skin formulation of the present invention include tromethamine. When used, the amount of pH adjuster is approximately 0.1 to approximately 2% by weight, preferably approximately 0.1 to approximately 1% by weight, based on the total weight of the composition.

[0050] In one specific embodiment of the invention, the topical skin agent comprises an acrylate / C10-30 alkanol acrylate crosspolymer, glycerin, p-hydroxyacetophenone, glyceryl stearate and lecithin, cetearyl alcohol, cetearyl ethylhexanoate, tromethamine or a combination thereof.

[0051] In some embodiments of the present invention, the amount of the plant oil composition in the topical skin preparation is 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).

[0052] Example

[0053] The present invention will be further illustrated below with reference to specific embodiments. It is necessary to point out that the scope of the present invention is not limited to these embodiments, which are intended to illustrate the technical solutions of the present invention and do not constitute a limitation on the scope of the present invention. Experimental methods in the following embodiments that do not specify specific conditions are generally performed under conventional conditions or as recommended by the manufacturer. The percentages of artemisia oil and safflower seed oil in the following comparative examples and embodiments are calculated by volume.

[0054] 1. Experimental Materials

[0055] Safflower (CARTHAMUS TINCTORIUS) seed oil, trade name: Safflower Seed Oil, purchased from Northstar Lipids UK Ltd.;

[0056] Artemisia annua oil was purchased from Shanghai Jahwa Biotechnology Co., Ltd.

[0057] 2. Instruments and Reagents

[0058] Instruments: benchtop centrifuge, HERAcell 240i CO2 incubator, Multiskan Mk3 (Thermo) microplate reader, 1300SERIES A2 biosafety cabinet, etc.

[0059] Reagents: MTT assay kit (Sigma), DMEM high glucose medium (Gibco 11995065), FBS, PBS (Solepro), dimethyl sulfoxide (DMSO, Sigma).

[0060] 3. Reagent preparation:

[0061] DMEM complete medium: Add FBS to DMEM high sugar medium to make the content 10%, and store at 4℃.

[0062] Comparative Example 1: Cell culture medium

[0063] According to calculations, the amount of DMSO added in each embodiment does not exceed 0.1%. Combined with past data, DMSO added at less than 0.1% does not have cytotoxicity. Therefore, for Comparative Example 1, DMEM complete medium with a DMSO content of 0.1% was selected for the experiment.

[0064] Comparative Example 1 cell culture medium: 1 ml of Comparative Example 1 cell culture medium = 1 μl DMSO + 999 μl DMEM complete culture medium, to obtain the culture medium of Comparative Example 1 with DMSO content of 0.01%.

[0065] Example 1: Cell culture medium

[0066] Mother liquor preparation: Artemisia annua oil and DMSO are prepared at a volume ratio of 1:499.

[0067] Example 1 Cell Culture Medium: 1 ml of Example 1 cell culture medium = 0.625 μl stock solution + 999.375 μl DMEM complete culture medium, to obtain the culture medium of Example 1 with an artemisia oil content of 0.000125%.

[0068] Example 2: Cell culture medium

[0069] Mother liquor preparation: Safflower seed oil and DMSO are prepared at a volume ratio of 1:1.

[0070] Example 2 Cell Culture Medium: 1 ml of Example 2 cell culture medium = 0.25 μl stock solution + 999.75 μl DMEM complete culture medium, to obtain the culture medium of Example 2 with a safflower seed oil content of 0.0125%.

[0071] Example 3: Cell culture medium

[0072] Mother liquor preparation: Safflower seed oil and DMSO are prepared at a volume ratio of 1:1.

[0073] Example 3 Cell Culture Medium: 1 ml of Example 3 cell culture medium = 0.5 μl stock solution + 999.5 μl DMEM complete culture medium, to obtain the culture medium of Example 3 with a safflower seed oil content of 0.025%.

[0074] Example 4: Cell culture medium

[0075] Mother liquor preparation: Safflower seed oil and DMSO are prepared at a volume ratio of 1:1.

[0076] Example 4 Cell Culture Medium: 1 ml of Example 4 cell culture medium = 1 μl stock solution + 999 μl DMEM complete culture medium, to obtain the culture medium of Example 4 with a safflower seed oil content of 0.05%.

[0077] Example 5: Cell culture medium

[0078] Mother liquor preparation: Safflower seed oil and DMSO are prepared at a volume ratio of 1:1.

[0079] Example 5 Cell Culture Medium: 1 ml of Example 5 cell culture medium = 2 μl stock solution + 998 μl DMEM complete culture medium, to obtain the culture medium of Example 5 with a safflower seed oil content of 0.1%.

[0080] Example 6: Cell culture medium

[0081] Mother liquor preparation: Artemisia annua oil: safflower seed oil: DMSO are prepared in a volume ratio of 1:100:99.

[0082] Example 6 Cell culture medium: 1 ml of Example 1 cell culture medium = 0.25 μl stock solution + 999.75 μl DMEM complete culture medium, to obtain the culture medium of Example 6 with an artemisia annua oil content of 0.000125% and safflower seed oil content of 0.125%.

[0083] Example 7 Cell Culture Medium:

[0084] Mother liquor preparation: Artemisia annua oil: safflower seed oil: DMSO are prepared in a volume ratio of 1:200:199.

[0085] Example 7 Cell Culture Medium: 1 ml of Example 1 cell culture medium = 0.5 μl stock solution + 999.5 μl DMEM complete culture medium, to obtain the culture medium of Example 7 with an artemisia annua oil content of 0.000125% and safflower seed oil content of 0.025%.

[0086] Example 8: Cell culture medium

[0087] Mother liquor preparation: Artemisia annua oil: safflower seed oil: DMSO were prepared in a volume ratio of 0.5:200:199.5.

[0088] Example 8 Cell culture medium: 1 ml of Example 1 cell culture medium = 1 μl stock solution + 999 μl DMEM complete culture medium, to obtain the culture medium of Example 8 with an artemisia annua oil content of 0.000125% and safflower seed oil content of 0.05%.

[0089] Example 9 Cell Culture Medium:

[0090] Mother liquor preparation: Artemisia annua oil: safflower seed oil: DMSO were prepared in a volume ratio of 0.5:400:399.5.

[0091] Example 9 Cell culture medium: 1 ml of Example 1 cell culture medium = 2 μl stock solution + 998 μl DMEM complete culture medium, to obtain the culture medium of Example 9 with an artemisia annua oil content of 0.000125% and safflower seed oil content of 0.1%.

[0092] Table 1 summarizes the content and ratio of artemisia annua oil and safflower seed oil per milliliter of culture medium in the examples.

[0093] Table 1

[0094]

[0095] Test Example 1: Keratinocyte Viability Test of Comparative Example 1 and Examples 1, 2, and 6

[0096] This invention employs a Hacat assay method for testing cell viability in immortalized human keratinocytes. An appropriate amount of the sample prepared in the examples is added to the keratinocytes and incubated for 24 hours. After 24 hours, the sample is washed away, and cell viability is tested using the MTT assay. Based on the increase in viability, the ability of the sample in promoting cell proliferation and cell viability can be evaluated.

[0097] Hacat cell seeding plate

[0098] Discard the culture medium in the T175 culture flask, wash once with PBS, add trypsin containing 0.25% EDTA for 1 min, then add fresh culture medium to stop the digestion. Transfer the cell suspension to a 15 mL centrifuge tube, centrifuge at 800 rpm for 5 min, and discard the supernatant. Resuspend in DMEM complete medium and count the cells. Dilute the cell suspension to a density of 8000 cells / 100 μL and seed 100 μL / well in 96-well plates. After seeding, incubate at 37°C, 5% CO2 for 24 h.

[0099] Cell processing and sample addition

[0100] Remove the 96-well plate, discard the supernatant, add 100 μL of the culture medium corresponding to Comparative Example 1 and Examples 2, 3 and 6, and incubate at 37°C and 5% CO2 for 24 h.

[0101] Cell proliferation and viability test

[0102] MTT stock solution preparation: Dissolve 25mg MTT in 5ml of MTT solvent to prepare a 5mg / ml MTT solution.

[0103] MTT solution preparation: Prepare according to the ratio of 10 μl MTT solution to 90 μl DMEM complete culture medium.

[0104] Add sample: Remove the original culture medium, add 100 μl of the above MTT solution to each well, and continue incubation in the cell culture incubator for 4 hours.

[0105] Dissolution: Carefully remove the MTT solution, add 100 μL of DMSO to each well and mix well. Continue incubation in a cell culture incubator until the crystalline, deep purple formazan product is completely dissolved and observed under a regular optical microscope.

[0106] Absorbance measurement: The absorbance was measured at 570 nm using an ELISA reader.

[0107]

[0108] Result judgment

[0109] Comparative Example 1 was the untreated group, with a relative activity of 100%. Examples containing artemisia annua oil and safflower seed oil were compared with Comparative Example 1 and examples containing only equal amounts of artemisia annua oil and safflower seed oil. If the cell proliferation rate and cell activity were significantly higher than other groups, it was considered to have a better effect on promoting cell proliferation and activity, and also a better effect on revitalizing the skin and repairing damage.

[0110] Experimental results

[0111] As shown in Table 2, compared with using artemisia annua oil and safflower seed oil of the same concentration alone, the combination of artemisia annua oil and safflower seed oil at a volume ratio of 1:100 has a significant effect on enhancing cell proliferation and vitality.

[0112] Table 2

[0113]

[0114]

[0115] All the above p-value calculations are compared with Example 6.

[0116] Test Example 2: Keratinocyte Viability Test of Comparative Example 1 and Examples 1, 3, and 7

[0117] The same experimental procedures as in Test Example 1 were used for the test.

[0118] Experimental results:

[0119] As shown in Table 3, compared with using artemisia annua oil and safflower seed oil of the same concentration alone, the combination of artemisia annua oil and safflower seed oil at a volume ratio of 1:200 has a significant effect on enhancing cell proliferation and vitality.

[0120] Table 3

[0121]

[0122] All the above p-value calculations are compared with Example 7.

[0123] Test Example 3: Keratinocyte Viability Test of Comparative Example 1 and Examples 1, 4, and 8

[0124] The same experimental procedures as in Test Example 1 were used for the test.

[0125] Experimental results:

[0126] As shown in Table 4, compared with using artemisia annua oil and safflower seed oil alone at the same concentration, the combination of artemisia annua oil and safflower seed oil at a volume ratio of 1:400 has a significant effect on enhancing cell proliferation and vitality.

[0127] Table 4

[0128]

[0129] All the above p-value calculations are compared with Example 8.

[0130] Test Example 4: Keratinocyte Viability Test of Comparative Example 1 and Examples 1, 5, and 9

[0131] The same experimental procedures as in Test Example 1 were used for the test.

[0132] Experimental results:

[0133] As shown in Table 5, compared with using artemisia annua oil and safflower seed oil of the same concentration alone, the combination of artemisia annua oil and safflower seed oil at a volume ratio of 1:800 has a significant effect on enhancing cell proliferation and vitality.

[0134] Table 5

[0135]

[0136]

[0137] All the above p-value calculations are compared with Example 9.

[0138] The plant oil compositions described in this invention can be used as functional additives in topical skin agents, preferably cosmetic compositions, including but not limited to the preparation of products in dosage forms such as creams, lotions, gels, toners, serums, masks, eye creams, aerosols (cleansing foams), sprays, shower gels, and facial cleansers. For example, the plant oil compositions of Examples 6, 7, 8, and 9 are used in topical skin agents at a weight percentage of 0.0001%-20% (w / w). A preferred weight percentage is 0.001%-10% (w / w). A more preferred weight percentage is 0.001%-5% (w / w). The most preferred weight percentage is 0.01%-5% (w / w).

[0139] The following are specific examples of the application of the plant oil compositions in topical skin preparations, as well as the formulations and preparation methods of these dosage forms. Specific applications are as follows:

[0140] Application Example 1: Preparation of Face Cream

[0141]

[0142]

[0143] Application Example 2: Emulsion Preparation

[0144]

[0145] Application Example 3: Preparation of Eye Cream

[0146]

[0147] Application Example 4: Preparation of Facial Masks

[0148]

[0149]

[0150] Application Example 5: Preparation of Serum

[0151]

[0152]

Claims

1. A plant oil composition for promoting cell proliferation and vitality, the plant oil composition comprising artemisia annua oil and safflower seed oil, wherein the volume ratio of artemisia annua oil to safflower seed oil is 1:100 to 1:800, wherein the artemisia annua oil is obtained by supercritical extraction of Artemisia annua, and wherein the safflower seed oil contains 50-90% by weight of linoleic acid.

2. The vegetable oil composition according to claim 1, wherein, The safflower seed oil contains 6% by weight of saturated fatty acids, 21% by weight of oleic acid and 73% by weight of linoleic acid.

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