Fibroblast compound liquid for skin care and preparation method of fibroblast compound liquid

Abstract

The invention discloses a fibroblast compound liquid for skin care and a preparation method thereof, and relates to the technical field of biological cosmetics. The fibroblast compound liquid disclosed by the invention is prepared from fibroblast liquid, cordyceps sobolifera spore oil, a celastrus orbiculatus extract, a hemp extract, oridonin, L-aspartic acid, hydroxyethyl cellulose and water. The components in the fibroblast compound liquid supplement each other, the fibroblast compound liquid is high in absorptivity, and skin elasticity and compactness are improved by stimulating metabolism of skin cells and generation of collagen, so that striae gravidarum is reduced or eliminated.

Description

Technical Field

[0001] The invention relates to the technical field of biological cosmetics, and in particular to a fibroblast compound liquid for skin care and a preparation method thereof. Background Art

[0002] The formation of stretch marks is mainly due to the influence of hormones during pregnancy, and the bulging abdomen damages or breaks the elastic fibers and collagen fibers of the skin, causing the abdominal skin to become thinner and thinner, and some pink or purple wavy patterns of different widths and lengths appear. After delivery, these patterns will gradually disappear, leaving behind white or silvery white shiny scar lines, i.e. stretch marks. Stretch marks mainly appear on the abdominal wall, and may also appear on the inner and outer thighs, buttocks, chest, lower back and arms. They are most obvious in primiparas. Once stretch marks appear, they are difficult to disappear, and are accompanied by sagging skin, sagging breasts, and abdominal fat accumulation, which seriously affects women's body shape and physical and mental health after childbirth. The current solution is basically to apply olive oil, lotion and other preventive measures during pregnancy, or to use all-trans retinoic acid or fruit acid compounds locally, or to use laser means to eliminate and dilute stretch marks, but the effect is not good. Skin care products made entirely of chemical raw materials are imperfect, manifested in unscientific formulations or containing substances that are harmful to the human body, such as containing a large amount of phosphorus, which will hinder the body's absorption of calcium, an important substance for the human body. For example, existing skin care products contain acrylamide, which is highly toxic to the nervous system and is a carcinogen; and mercury-containing compounds are commonly used as whitening agents and preservatives in skin care products, which may cause skin spots, red blood streaks, allergies and other problems, and their efficacy is not satisfactory. Products formulated with plant extracts are naturally uncertain in their efficacy due to the simple extraction method and the complex and uncertain ingredients in the crude extract. Moreover, most existing skin care products do not have biologically active substances and cannot promote skin metabolism or improve stretch marks.

[0003] Fibroblasts are a kind of matrix cells, which are widely present in the connective tissues of humans and animals. The bioactive components in the fibroblast fluid can be used to achieve good skin care effects, and its production method is separated from the chemical production process and has good safety. Application No. CN201310345593.0 discloses a fibroblast fluid for beauty care and a preparation method thereof, and the fibroblast fluid contains cell culture fluid, collagen, cell growth factor, amino acid and vitamin, but its effect data is unclear and the operation is complicated. Therefore, it is urgent to provide a simple and efficient fibroblast compound solution for skin care and a preparation method thereof. Summary of the invention

[0004] In view of the above-mentioned defects of the prior art, the present invention proposes a fibroblast compound solution for skin care and a preparation method thereof to solve the problems raised by the above-mentioned background technology.

[0005] In order to achieve the above object, the present invention provides the following technical solutions:

[0006] A fibroblast compound liquid for skin care comprises, by weight, 10 to 20 parts of fibroblast liquid, 10 to 15 parts of cicada fungus spore oil, 7 to 13 parts of orbiculatus extract, 6 to 12 parts of potentilla extract, 3 to 7 parts of oridonin, 2 to 6 parts of L-aspartic acid, 1 to 5 parts of hydroxyethyl cellulose and 40 to 60 parts of water.

[0007] Preferably, a fibroblast complex solution for skin care comprises, by weight, 15 parts of fibroblast solution, 12 parts of cicada fungus spore oil, 10 parts of celastrus orbiculatus extract, 10 parts of potentilla anserina extract, 6 parts of oridonin, 5 parts of L-aspartic acid, 4 parts of hydroxyethyl cellulose and 50 parts of water.

[0008] Preferably, the fibroblasts in the fibroblast fluid are HSF human skin fibroblasts, derived from IMMOCELL, item number IM-H032, fibroblast basal culture medium (self-use DMEM / F12+10% FBS+1% p / s+0.005mg / ml insulin+5ng / ml BfGF+1ug / ml hydrocortisone+50ug / ml ascorbic acid (vitamin C)+7.5mM L-Gln). After receiving the cells, wipe the surface of the cell bottle with 75% alcohol, place it in a 37-degree incubator and culture it for 2 to 4 hours to stabilize the cell state. When the cell density reaches 80% to 90%, subculture can be carried out.

[0009] Subculture: a. Discard the culture supernatant and rinse the cells 1-2 times with PBS without calcium and magnesium ions. b. Add 1mL of digestion solution (0.25% Trypsin-0.02% EDTA) to the culture bottle, so that the digestion solution infiltrates all cells, place the culture bottle in a 37℃ incubator for digestion for 1-2 minutes (depending on the condition of the cells), and then observe the cell digestion under a microscope. If most of the cells become round and fall off, quickly take it back to the operating table, tap the culture bottle a few times, and add 2-3ml of complete culture medium to terminate the digestion. After gently mixing, put it into a sterile centrifuge tube, centrifuge at 1000rpm for 5 minutes, discard the supernatant, add 1-2mL of culture solution and blow it evenly. c. Divide the cell suspension into a new dish or bottle containing 8mL of culture medium at a ratio of 1:2, and place it in an incubator for culture. d. Collect the cell supernatant in the culture dish that has been subcultured to the 4th to 6th generation to obtain fibroblast fluid.

[0010] Preferably, the preparation method of the Cicadae Convolvulus spore oil comprises the following steps:

[0011] S1. First, the spore powder of Cicadae Convolvulus is uniformly mixed with anhydrous ethanol at a ratio of 1:(12-20) g / mL, and ultrasonically treated to form a treatment solution;

[0012] S2, freezing the treated liquid with liquid nitrogen and then rewarming it to a temperature of 20-26° C., repeating this process 3-5 times, and then placing it in an extraction kettle, extracting it with subcritical propane to obtain an extract;

[0013] S3. The extract is desolventized and separated for 10 to 20 minutes at a temperature of 50 to 60° C. and a pressure of -0.5 to -0.3 MPa. After propane separation, the extract is returned to the solvent tank, the cicada fungus spore mixed oil is collected, and anhydrous ethanol is separated by reduced pressure distillation. The cicada fungus spore oil is obtained by further treatment.

[0014] Preferably, the frequency of the ultrasonic treatment in step S1 is 20 to 35 KHz, and the time is 3 to 5 hours.

[0015] Preferably, the extraction step in step S2 is as follows: the extraction temperature is 35-45°C, when the pressure of the extraction kettle rises to 0.3-0.5MPa, it is maintained for 5-8min, and then the pressure is released to 0.1MPa; thereafter, the pressure is continuously increased to 0.3-0.5MPa and maintained for 2-4h.

[0016] Preferably, the further processing in step S3 comprises adding an equal volume ratio of Antarctic krill oil to the oil.

[0017] Preferably, the method for preparing the Celastrus orbiculatus extract comprises the following steps:

[0018] ① Dry and crush the roots and stems of Celastrus orbiculatus to obtain Celastrus orbiculatus powder;

[0019] ② adding ursolic acid with a mass concentration of 15-25% to the Celastrus orbiculatus powder at a material-liquid ratio of 1:(5-10) g / mL to obtain a premix;

[0020] ③ Filter the premix to obtain a filtrate, and emulsify the filtrate at a speed of 3000-4000 r / min and a temperature of 25-30° C. for 40-50 minutes to obtain an emulsion;

[0021] ④ Add tannic acid aqueous solution with a mass concentration of 2-5% to the emulsion, let it stand for 100-200 minutes and then dry it at a temperature of 60-80° C. to obtain the Celastrus orbiculatus extract.

[0022] Preferably, in step ④, the volume ratio of the emulsion to the tannic acid aqueous solution is 1:(1.5-3).

[0023] Preferably, the preparation method of the Potentilla anserine extract comprises the following steps: taking the whole plant of Potentilla anserine, drying and then crushing it, mixing it with 70-80% ethanol by volume at a solid-liquid ratio of 1:(3-5) g / mL, refluxing extraction for 60-100 minutes, collecting the extract, and concentrating it by reduced pressure evaporation to obtain the Potentilla anserine extract.

[0024] Preferably, a method for preparing a fibroblast compound solution for skin care comprises the following steps:

[0025] (1) mixing fibroblast fluid, Rubescensine A and L-aspartic acid uniformly to prepare liquid A;

[0026] (2) heating water to 60-70° C., adding Celastrus orbiculatus extract and Potentilla anserina extract, stirring evenly, cooling to 30-40° C., and then adding Cicadae confusa spore oil to prepare Liquid B;

[0027] (3) Mix solution A and solution B, stir evenly, then add hydroxyethyl cellulose, stir and cool to 22-26° C. to obtain a fibroblast compound solution.

[0028] Compared with the prior art, the present invention has the following beneficial effects:

[0029] (1) The fibroblast complex solution for skin care provided by the present invention comprises fibroblast solution that can secrete collagen, elastin and hyaluronic acid. Collagen and elastin together give the skin firmness and elasticity, support the epidermis and subcutaneous tissue, and maintain the elasticity and firmness of the skin. Hyaluronic acid can control the cells to contain a large amount of water at all times, maintain the epidermis moisturizing, and also plays an important role in moisturizing and nourishing the skin, and can effectively prevent stretch marks or improve the symptoms of stretch marks.

[0030] (2) The fibroblast compound liquid for skin care prepared by the present invention is added with Cicadae Constricta spore oil, Celastrus orbiculatus extract, Potentilla anserina extract and Rubescensine A, which are all natural plant ingredients. The main ingredients of Cicadae Constricta spores include triterpenoid ganoderic acid, unsaturated fatty acids and organic germanium. These ingredients give Cicadae Constricta spore oil unique biological activity and health care functions. Triterpenoid ganoderic acid is a kind of unsaturated fatty acid, which has multiple biological activities such as anti-oxidation, anti-inflammatory and anti-tumor, and plays an important role in maintaining human health. In addition, Cicadae Constricta spore oil also contains organic germanium, which is a trace element beneficial to the human body, can enhance the body's immunity, resist oxidative stress, and help maintain physical health; the active ingredients of Celastrus orbiculatus extract include celangulin, celangulin, lignon, triterpenoids, etc., which have obvious anti-inflammatory effects, can inhibit the migration and activation of inflammatory cells, reduce the production of inflammatory factors, and thus reduce the redness, swelling and itching of stretch marks. Potentilla extract contains a variety of active substances, including antioxidants, α-glucosidase inhibitors, tyrosinase inhibitors, triterpenoids, flavonoids, etc., which have immunomodulatory, anti-inflammatory and antibacterial activities; Rubescensine A belongs to the enantio-kaurene type tetracyclic diterpenoid compound, which shows significant anti-inflammatory effects. By adding a variety of natural plant ingredients, it can effectively improve the formation of stretch marks.

[0031] (3) The raw materials used in the fibroblast complex solution are all natural plant ingredients, do not contain heavy metals, antibiotics and hormones, and will not produce toxic side effects, ensuring the safety of the product. Through the reasonable combination of various ingredients, the effective ingredients contained can be quickly introduced into the dermis of the skin to accelerate skin repair, effectively repair the skin collagen fibers and elastic fibers that form stretch marks, and at the same time lighten the pigment.

[0032] (4) The preparation method of cicada fungus spore oil provided by the present invention has a high yield of spore oil. In the extraction process, the pressure of the extraction kettle is first increased to 0.3-0.5 MPa, maintained for 5-8 minutes, and then the pressure is released to 0.1 MPa. During the pressure increase and pressure release process, the solvent will immerse and swell in the cicada fungus spores, causing damage to their tissues, which helps to improve the mass transfer and dissolution of subsequent oil components. At the same time, in order to increase the stability of cicada fungus spore oil, Antarctic krill oil is added to the oil to prevent product stratification and improve stability. The acid extraction of the orbiculatus extract has high efficacy. The present invention provides new uses of cicada fungus spore oil, orbiculatus extract, pothopica extract and oridonin in preventing and eliminating stretch marks. DETAILED DESCRIPTION

[0033] The technical solution of the present invention is further described in detail below in conjunction with specific implementation methods.

[0034] Example 1

[0035] A fibroblast compound liquid for skin care comprises, by weight, 10 parts of fibroblast liquid, 10 parts of cicada fungus spore oil, 7 parts of orbiculatus extract, 6 parts of potentilla extract, 3 parts of oridonin, 2 parts of L-aspartic acid, 1 part of hydroxyethyl cellulose and 40 parts of water.

[0036] The preparation method of the cicada condy spore oil comprises the following steps:

[0037] S1, firstly, the spore powder of Cicadae Convolvulus and anhydrous ethanol were uniformly mixed at a solid-liquid ratio of 1:12 g / mL, and ultrasonic treatment was performed to form a treatment liquid; wherein the frequency of the ultrasonic treatment was 20 KHz, and the time was 3 h;

[0038] S2, freezing the treated liquid with liquid nitrogen and rewarming it to a temperature of 20°C, repeating this process three times, and then placing it in an extraction kettle, extracting it with subcritical propane to obtain an extract; wherein the extraction steps are: the extraction temperature is 35°C, when the pressure of the extraction kettle rises to 0.3MPa, it is maintained for 5 minutes, and then the pressure is released to 0.1MPa; then the pressure is continuously raised to 0.3MPa and maintained for 2 hours;

[0039] S3. The extract is desolventized and separated at a temperature of 50°C and a pressure of -0.5MPa for 10 minutes. After propane separation, it is returned to the solvent tank, and the cicada fungus spore mixed oil is collected. Anhydrous ethanol is separated by reduced pressure distillation at a pressure of 1.5kpa and a temperature of 40°C. Antarctic krill oil in an equal volume ratio is added to the oil to obtain cicada fungus spore oil.

[0040] The preparation method of the Celastrus orbiculatus extract comprises the following steps:

[0041] ① Dry and crush the roots and stems of Celastrus orbiculatus to obtain Celastrus orbiculatus powder;

[0042] ② adding ursolic acid with a mass concentration of 15% to the Celastrus orbiculatus powder at a material-liquid ratio of 1:5 g / mL to obtain a premix;

[0043] ③ Filter the premix to obtain a filtrate, and emulsify the filtrate at a speed of 3000 r / min and a temperature of 25° C. for 40 minutes to obtain an emulsion;

[0044] ④ Add a 2% mass concentration of tannic acid aqueous solution to the emulsion, let it stand for 100 minutes and then dry it at 60°C to obtain the Celastrus orbiculatus extract; wherein the volume ratio of the emulsion to the tannic acid aqueous solution is 1:1.5.

[0045] The preparation method of the Potentilla anserine extract comprises the following steps: taking the whole plant of Potentilla anserine, drying it and then crushing it, mixing it with 70% ethanol by volume at a solid-liquid ratio of 1:3 g / mL, refluxing and extracting it for 60 minutes, collecting the extract, and concentrating it by reduced pressure evaporation to obtain the Potentilla anserine extract.

[0046] A method for preparing a fibroblast compound solution for skin care, comprising the following steps:

[0047] (1) mixing fibroblast fluid, Rubescensine A and L-aspartic acid uniformly to prepare liquid A;

[0048] (2) heating water to 60° C., adding Celastrus orbiculatus extract and Pteridium anserine extract, stirring evenly, cooling to 30° C., and then adding Cicadae confusa spore oil to prepare liquid B;

[0049] (3) Mix solution A and solution B, stir evenly, then add hydroxyethyl cellulose, stir and cool to 22° C. to obtain a fibroblast compound solution.

[0050] Example 2

[0051] A fibroblast compound liquid for skin care comprises, by weight, 20 parts of fibroblast liquid, 15 parts of cicada fungus spore oil, 13 parts of orbiculatus extract, 12 parts of potentilla extract, 7 parts of oridonin, 6 parts of L-aspartic acid, 5 parts of hydroxyethyl cellulose and 60 parts of water.

[0052] The preparation method of the cicada condy spore oil comprises the following steps:

[0053] S1, first cicada conidia spore powder and anhydrous ethanol are uniformly mixed at a solid-liquid ratio of 1:20g / mL, and ultrasonic treatment is performed to form a treatment liquid; wherein the frequency of the ultrasonic treatment is 35KHz, and the time is 5h;

[0054] S2, freezing and reheating the treated liquid to 26°C with liquid nitrogen, repeating 5 times, and then placing it in an extraction kettle, extracting it with subcritical propane to obtain an extract; wherein the extraction steps are: the extraction temperature is 45°C, when the pressure of the extraction kettle rises to 0.5MPa, it is maintained for 8 minutes, and then the pressure is released to 0.1MPa; then the pressure is continuously increased to 0.5MPa and maintained for 4 hours;

[0055] S3. The extract is desolventized and separated at a temperature of 60°C and a pressure of -0.3MPa for 20 minutes. After propane separation, it returns to the solvent tank, collects the cicada fungus spore mixed oil, and separates the anhydrous ethanol by reduced pressure distillation at a pressure of 1.5kpa and a temperature of 40°C. Antarctic krill oil in an equal volume ratio is added to the oil to obtain cicada fungus spore oil.

[0056] The preparation method of the Celastrus orbiculatus extract comprises the following steps:

[0057] ① Dry and crush the roots and stems of Celastrus orbiculatus to obtain Celastrus orbiculatus powder;

[0058] ② adding ursolic acid with a mass concentration of 25% to the Celastrus orbiculatus powder at a material-liquid ratio of 1:10 g / mL to obtain a premix;

[0059] ③ Filter the premix to obtain a filtrate, and emulsify the filtrate at a speed of 4000 r / min and a temperature of 30° C. for 50 minutes to obtain an emulsion;

[0060] ④ Add a 5% mass concentration of tannic acid aqueous solution to the emulsion, let it stand for 200 minutes and then dry it at 80°C to obtain the Celastrus orbiculatus extract; wherein the volume ratio of the emulsion to the tannic acid aqueous solution is 1:3.

[0061] The preparation method of the Potentilla anserine extract comprises the following steps: taking the whole plant of Potentilla anserine, drying and then crushing it, mixing it with 80% ethanol by volume at a solid-liquid ratio of 1:5 g / mL, refluxing extraction for 100 minutes, collecting the extract, and concentrating it under reduced pressure evaporation at a pressure of 1.5 kpa and a temperature of 40° C. to obtain the Potentilla anserine extract.

[0062] A method for preparing a fibroblast compound solution for skin care, comprising the following steps:

[0063] (1) mixing fibroblast fluid, Rubescensine A and L-aspartic acid uniformly to prepare liquid A;

[0064] (2) heating water to 70° C., adding Celastrus orbiculatus extract and Potentilla anserina extract, stirring evenly, cooling to 40° C., and then adding Cicadae confusa spore oil to prepare Solution B;

[0065] (3) Mix solution A and solution B, stir evenly, then add hydroxyethyl cellulose, stir and cool to 26° C. to obtain a fibroblast compound solution.

[0066] Example 3

[0067] A fibroblast compound liquid for skin care comprises, by weight, 15 parts of fibroblast liquid, 12 parts of cicada fungus spore oil, 10 parts of orbiculatus extract, 10 parts of potentilla extract, 6 parts of oridonin, 5 parts of L-aspartic acid, 4 parts of hydroxyethyl cellulose and 50 parts of water.

[0068] The preparation method of the cicada condy spore oil comprises the following steps:

[0069] S1, first cicada conidia spore powder and anhydrous ethanol are uniformly mixed at a solid-liquid ratio of 1:18g / mL, and ultrasonic treatment is performed to form a treatment liquid; wherein the frequency of the ultrasonic treatment is 30KHz, and the time is 4h;

[0070] S2, freezing and reheating the treated liquid to 22°C with liquid nitrogen, repeating 4 times, and then placing it in an extraction kettle, extracting it with subcritical propane to obtain an extract; wherein the extraction steps are: the extraction temperature is 40°C, when the pressure of the extraction kettle rises to 0.4MPa, it is maintained for 7min, and then the pressure is released to 0.1MPa; then the pressure is continuously increased to 0.4MPa and maintained for 3h;

[0071] S3. The extract is desolventized and separated for 15 minutes at a temperature of 55°C and a pressure of -0.4MPa. After propane separation, it is returned to the solvent tank, and the cicada fungus spore mixed oil is collected. Anhydrous ethanol is separated by reduced pressure distillation at a pressure of 1.5kpa and a temperature of 40°C. Antarctic krill oil in an equal volume ratio is added to the oil to obtain cicada fungus spore oil.

[0072] The preparation method of the Celastrus orbiculatus extract comprises the following steps:

[0073] ① Dry and crush the roots and stems of Celastrus orbiculatus to obtain Celastrus orbiculatus powder;

[0074] ② Adding ursolic acid with a mass concentration of 20% to the Celastrus orbiculatus powder at a material-liquid ratio of 1:7 g / mL to obtain a premix;

[0075] ③ Filter the premix to obtain a filtrate, and emulsify the filtrate at a speed of 3500 r / min and a temperature of 27° C. for 45 minutes to obtain an emulsion;

[0076] ④ Add a 3% mass concentration of tannic acid aqueous solution to the emulsion, let it stand for 150 minutes and then dry it at 70°C to obtain the Celastrus orbiculatus extract; wherein the volume ratio of the emulsion to the tannic acid aqueous solution is 1:2.

[0077] The preparation method of the Potentilla anserine extract comprises the following steps: taking the whole plant of Potentilla anserine, drying and then crushing it, mixing it with 75% ethanol by volume at a solid-liquid ratio of 1:4 g / mL, refluxing extraction for 80 minutes, collecting the extract, and concentrating it by reduced pressure evaporation at a pressure of 1.5 kpa and a temperature of 40° C. to obtain the Potentilla anserine extract.

[0078] A method for preparing a fibroblast compound solution for skin care, comprising the following steps:

[0079] (1) mixing fibroblast fluid, Rubescensine A and L-aspartic acid uniformly to prepare liquid A;

[0080] (2) heating water to 65° C., adding Celastrus orbiculatus extract and Potentilla anserina extract, stirring evenly, cooling to 35° C., and then adding Cicadae confusa spore oil to prepare Solution B;

[0081] (3) Mix solution A and solution B, stir evenly, then add hydroxyethyl cellulose, stir and cool to 25° C. to obtain a fibroblast compound solution.

[0082] Comparative Example 1

[0083] The difference between this comparative example and Example 3 is that the cicada fungus spore oil is replaced by Ganoderma lucidum spore oil, and the other ingredients and steps remain unchanged.

[0084] A fibroblast compound liquid for skin care comprises, by weight, 15 parts of fibroblast liquid, 12 parts of ganoderma lucidum spore oil, 10 parts of orbiculatus extract, 10 parts of potentilla extract, 6 parts of oridonin, 5 parts of L-aspartic acid, 4 parts of hydroxyethyl cellulose and 50 parts of water.

[0085] The preparation method of the ganoderma lucidum spore oil comprises the following steps:

[0086] S1. First, the Ganoderma lucidum spore powder and anhydrous ethanol were uniformly mixed at a solid-liquid ratio of 1:18 g / mL, and ultrasonic treatment was performed to form a treatment liquid; wherein the frequency of the ultrasonic treatment was 30 KHz and the time was 4 h;

[0087] S2, freezing the treated liquid with liquid nitrogen and then rewarming it to 22°C, repeating this process 4 times, and then placing it in an extraction kettle, extracting it with subcritical propane to obtain an extract; wherein the extraction steps are: the extraction temperature is 40°C, when the pressure of the extraction kettle rises to 0.4MPa, it is maintained for 7 minutes, and then the pressure is released to 0.1MPa; then the pressure is continuously raised to 0.4MPa and maintained for 3 hours;

[0088] S3. The extract is desolventized and separated for 15 minutes at a temperature of 55°C and a pressure of -0.4MPa. After propane separation, it is returned to the solvent tank, and the Ganoderma lucidum spore mixed oil is collected. Anhydrous ethanol is separated by reduced pressure distillation at a pressure of 1.5kpa and a temperature of 40°C. Antarctic krill oil in an equal volume ratio is added to the oil to obtain Ganoderma lucidum spore oil.

[0089] A method for preparing a fibroblast compound solution for skin care, comprising the following steps:

[0090] (1) mixing fibroblast fluid, Rubescensine A and L-aspartic acid uniformly to prepare liquid A;

[0091] (2) heating water to 65° C., adding Celastrus orbiculatus extract and Potentilla anserina extract, stirring evenly, cooling to 35° C., and then adding Ganoderma lucidum spore oil to prepare liquid B;

[0092] (3) Mix solution A and solution B, stir evenly, then add hydroxyethyl cellulose, stir and cool to 25° C. to obtain a fibroblast compound solution.

[0093] Comparative Example 2

[0094] The difference between this comparative example and Example 3 is that the preparation method of Cicadae Convolvulus spore oil does not use repeated liquid nitrogen freezing steps, and the other components and steps remain unchanged.

[0095] A fibroblast compound liquid for skin care comprises, by weight, 15 parts of fibroblast liquid, 12 parts of cicada fungus spore oil, 10 parts of orbiculatus extract, 10 parts of potentilla extract, 6 parts of oridonin, 5 parts of L-aspartic acid, 4 parts of hydroxyethyl cellulose and 50 parts of water.

[0096] The preparation method of the cicada condy spore oil comprises the following steps:

[0097] S1, first cicada conidia spore powder and anhydrous ethanol are uniformly mixed at a solid-liquid ratio of 1:18g / mL, and ultrasonic treatment is performed to form a treatment liquid; wherein the frequency of the ultrasonic treatment is 30KHz, and the time is 4h;

[0098] S2, placing the treated liquid in an extraction kettle, extracting with subcritical propane to obtain an extract; wherein the extraction steps are: the extraction temperature is 40°C, when the pressure of the extraction kettle rises to 0.4MPa, it is maintained for 7min, and then the pressure is released to 0.1MPa; then the pressure is continuously raised to 0.4MPa and maintained for 3h;

[0099] S3. The extract is desolventized and separated for 15 minutes at a temperature of 55°C and a pressure of -0.4MPa. After propane separation, it is returned to the solvent tank, and the cicada fungus spore mixed oil is collected. Anhydrous ethanol is separated by reduced pressure distillation at a pressure of 1.5kpa and a temperature of 40°C. Antarctic krill oil in an equal volume ratio is added to the oil to obtain cicada fungus spore oil.

[0100] Comparative Example 3

[0101] The difference between this comparative example and Example 3 is that the preparation method of cicada condyle spore oil does not add Antarctic krill oil, and the other ingredients and steps remain unchanged.

[0102] A fibroblast compound liquid for skin care comprises, by weight, 15 parts of fibroblast liquid, 12 parts of cicada fungus spore oil, 10 parts of orbiculatus extract, 10 parts of potentilla extract, 6 parts of oridonin, 5 parts of L-aspartic acid, 4 parts of hydroxyethyl cellulose and 50 parts of water.

[0103] The preparation method of the cicada condy spore oil comprises the following steps:

[0104] S1, first cicada conidia spore powder and anhydrous ethanol are uniformly mixed at a solid-liquid ratio of 1:18g / mL, and ultrasonic treatment is performed to form a treatment liquid; wherein the frequency of the ultrasonic treatment is 30KHz, and the time is 4h;

[0105] S2, freezing the treated liquid with liquid nitrogen and then rewarming it to 22°C, repeating this process 4 times, and then placing it in an extraction kettle, extracting it with subcritical propane to obtain an extract; wherein the extraction steps are: the extraction temperature is 40°C, when the pressure of the extraction kettle rises to 0.4MPa, it is maintained for 7 minutes, and then the pressure is released to 0.1MPa; then the pressure is continuously raised to 0.4MPa and maintained for 3 hours;

[0106] S3. The extract is desolventized and separated for 15 minutes at a temperature of 55°C and a pressure of -0.4MPa. After propane separation, it is returned to the solvent tank, and the cicada fungus spore mixed oil is collected. Anhydrous ethanol is separated by reduced pressure distillation at a pressure of 1.5kpa and a temperature of 40°C to obtain cicada fungus spore oil.

[0107] Comparative Example 4

[0108] The difference between this comparative example and Example 3 is that in the preparation method of Celastrus orbiculatus extract, ursolic acid is replaced with anhydrous ethanol, and the other components and steps remain unchanged.

[0109] A fibroblast compound liquid for skin care comprises, by weight, 15 parts of fibroblast liquid, 12 parts of cicada fungus spore oil, 10 parts of orbiculatus extract, 10 parts of potentilla extract, 6 parts of oridonin, 5 parts of L-aspartic acid, 4 parts of hydroxyethyl cellulose and 50 parts of water.

[0110] The preparation method of the Celastrus orbiculatus extract comprises the following steps:

[0111] ① Dry and crush the roots and stems of Celastrus orbiculatus to obtain Celastrus orbiculatus powder;

[0112] ② Adding anhydrous ethanol to the Celastrus orbiculatus powder at a material-liquid ratio of 1:7 g / mL to obtain a premix;

[0113] ③ Filter the premix to obtain a filtrate, and emulsify the filtrate at a speed of 3500 r / min and a temperature of 27° C. for 45 minutes to obtain an emulsion;

[0114] ④ Add a 3% mass concentration of tannic acid aqueous solution to the emulsion, let it stand for 150 minutes and then dry it at 70°C to obtain the Celastrus orbiculatus extract; wherein the volume ratio of the emulsion to the tannic acid aqueous solution is 1:2.

[0115] Comparative Example 5

[0116] The difference between this comparative example and Example 3 is that in the preparation method of Celastrus orbiculatus extract, the tannic acid aqueous solution is replaced by a citric acid aqueous solution, and the other components and steps remain unchanged.

[0117] A fibroblast compound liquid for skin care comprises, by weight, 15 parts of fibroblast liquid, 12 parts of cicada fungus spore oil, 10 parts of orbiculatus extract, 10 parts of potentilla extract, 6 parts of oridonin, 5 parts of L-aspartic acid, 4 parts of hydroxyethyl cellulose and 50 parts of water.

[0118] The preparation method of the Celastrus orbiculatus extract comprises the following steps:

[0119] ① Dry and crush the roots and stems of Celastrus orbiculatus to obtain Celastrus orbiculatus powder;

[0120] ② Adding ursolic acid with a mass concentration of 20% to the Celastrus orbiculatus powder at a material-liquid ratio of 1:7 g / mL to obtain a premix;

[0121] ③ Filter the premix to obtain a filtrate, and emulsify the filtrate at a speed of 3500 r / min and a temperature of 27° C. for 45 minutes to obtain an emulsion;

[0122] ④ Add 3% citric acid aqueous solution to the emulsion, let it stand for 150 minutes and then dry it at 70°C to obtain the Celastrus orbiculatus extract; wherein the volume ratio of the emulsion to the citric acid aqueous solution is 1:2.

[0123] Comparative Example 6

[0124] The difference between this comparative example and Example 3 is that no Potentilla anserina extract is added to the fibroblast compound solution, and the other ingredients and steps remain unchanged.

[0125] A fibroblast compound liquid for skin care comprises, by weight, 15 parts of fibroblast liquid, 12 parts of cicada fungus spore oil, 10 parts of orbiculatus extract, 6 parts of oridonin, 5 parts of L-aspartic acid, 4 parts of hydroxyethyl cellulose and 50 parts of water.

[0126] A method for preparing a fibroblast compound solution for skin care, comprising the following steps:

[0127] (1) mixing fibroblast fluid, Rubescensine A and L-aspartic acid uniformly to prepare liquid A;

[0128] (2) heating water to 65° C., adding Celastrus orbiculatus extract, stirring evenly, cooling to 35° C., and then adding Cicadae Convolvulus spore oil to prepare Solution B;

[0129] (3) Mix solution A and solution B, stir evenly, then add hydroxyethyl cellulose, stir and cool to 25° C. to obtain a fibroblast compound solution.

[0130] Comparative Example 7

[0131] The difference between this comparative example and Example 3 is that the preparation method of the fibroblast compound solution is changed, and the other components and steps remain unchanged.

[0132] A fibroblast compound liquid for skin care comprises, by weight, 15 parts of fibroblast liquid, 12 parts of cicada fungus spore oil, 10 parts of orbiculatus extract, 10 parts of potentilla extract, 6 parts of oridonin, 5 parts of L-aspartic acid, 4 parts of hydroxyethyl cellulose and 50 parts of water.

[0133] A method for preparing a fibroblast compound solution for skin care, comprising the following steps:

[0134] The fibroblast liquid, Rubescensine A, L-aspartic acid, Celastrus orbiculatus extract, Potentilla anserina extract, Cicadae confusa spore oil and water at a temperature of 65° C. are uniformly mixed, and hydroxyethyl cellulose is added to obtain a fibroblast compound liquid;

[0135] Experimental example: Stretch mark removal test

[0136] Experimental subjects: Healthy women who have given birth and have new stretch marks were selected as subjects. 110 volunteers aged 26 to 35 were divided into 11 groups. The fibroblast compound solutions prepared in Examples 1 to 3 and Comparative Examples 1 to 7, as well as commercially available stretch mark removal products (main ingredients: retinoic acid, bitter almond oil, olive oil, cocoa butter, vitamin E and glycerin) were used respectively.

[0137] Inclusion criteria: ① More than 3 months postpartum, stretch marks appeared 0-3 years ago, and there were obvious stretch marks around the abdomen; ② The patients agreed to this study and signed the informed consent form; ③ Those who had not received other stretch mark treatments in the past year and were willing to accept follow-up.

[0138] Exclusion criteria: Those with any of the following conditions were not included in this study: ① Known allergies to cosmetics, skin care products, topical drugs and their ingredients; ② Women who are still breastfeeding; ③ Abdominal inflammation, eczema and other skin abnormalities, and systemic diseases that will affect the research results;

[0139] Experimental method: The subjects divided the abdomen into two areas with the midline of the abdomen as the boundary. One side was randomly selected as the test product application site, and the other side was selected as the control site. A stretch mark of similar severity was selected on each side as a mark for clinical evaluation of the target skin lesions. Before use, scrub the abdomen with warm water, apply 10mL of the test product to the test site, and gently massage the skin for 20 minutes. Each subject used it once a day at 19:00, and the test period was 2 months. Clinical evaluation criteria: Evaluate the overall repair of stretch marks, stretch mark area, stretch mark color, and stretch mark relaxation. At the same time, a 10-point method was used for scoring (0-10 points), 10 points represent a perfect state, 0 points represent a severe state, and the average value is taken. The test results are shown in Table 1.

[0140] Table 1 Stretch mark removal test

[0141]

[0142]

[0143] As shown in Table 1, the fibroblast compound solution prepared in Examples 1 to 3 has the best effect in removing stretch marks, especially Example 3. As shown in Example 3 compared with Comparative Examples 1, 4 and 5, when the cicada condyle spore oil, ursolic acid and tannic acid aqueous solution are replaced with Ganoderma lucidum spore oil, anhydrous ethanol and citric acid aqueous solution, the overall repair effect of stretch marks is reduced, indicating that the technical solution has a certain selectivity for each component. Even though cicada condyle spore oil and Ganoderma lucidum spore oil have similarities in ingredients and efficacy, the specific effects may vary due to factors such as product manufacturing process and ingredient content. Ursolic acid can increase the efficacy of Celastrus orbiculatus, promote skin cell regeneration, and have a certain improvement effect on skin scars. Tannic acid aqueous solution can further increase the effect of removing stretch marks; as shown in Example 3 compared with Comparative Examples 2, 3 and 6, in the process of preparing cicada condyle spore oil, adding liquid nitrogen freezing and rewarming process can preserve The invention discloses a novel ...

[0144] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. A fibroblast compound solution for skin care, characterized in that: The invention comprises, by weight, 10 to 20 parts of fibroblast fluid, 10 to 15 parts of cicada fungus spore oil, 7 to 13 parts of orbiculatus scoparia extract, 6 to 12 parts of pothopilla extract, 3 to 7 parts of oridonin, 2 to 6 parts of L-aspartic acid, 1 to 5 parts of hydroxyethyl cellulose and 40 to 60 parts of water.

2. The fibroblast compound solution for skin care according to claim 1, characterized in that: The invention comprises, by weight, 15 parts of fibroblast fluid, 12 parts of cicada fungus spore oil, 10 parts of orbiculatus extract, 10 parts of pothopilla extract, 6 parts of oridonin, 5 parts of L-aspartic acid, 4 parts of hydroxyethyl cellulose and 50 parts of water.

3. The fibroblast complex solution for skin care according to claim 1, characterized in that: The preparation method of the cicada condy spore oil comprises the following steps: S1. First, the spore powder of Cicadae Convolvulus is uniformly mixed with anhydrous ethanol at a ratio of 1:(12-20) g / mL, and ultrasonically treated to form a treatment solution; S2, freezing the treated liquid with liquid nitrogen and then rewarming it to a temperature of 20-26° C., repeating this process 3-5 times, and then placing it in an extraction kettle, extracting it with subcritical propane to obtain an extract; S3. The extract is desolventized and separated for 10 to 20 minutes at a temperature of 50 to 60° C. and a pressure of -0.5 to -0.3 MPa. After propane separation, the extract is returned to the solvent tank, the cicada fungus spore mixed oil is collected, and anhydrous ethanol is separated by reduced pressure distillation. The cicada fungus spore oil is obtained by further treatment.

4. The fibroblast compound solution for skin care according to claim 3, characterized in that: The frequency of the ultrasonic treatment in step S1 is 20 to 35 KHz, and the time is 3 to 5 hours.

5. The fibroblast compound solution for skin care according to claim 3, characterized in that: The extraction step in step S2 is as follows: the extraction temperature is 35-45° C., when the pressure of the extraction kettle rises to 0.3-0.5 MPa, it is maintained for 5-8 minutes, and then the pressure is released to 0.1 MPa; thereafter, the pressure is continuously increased to 0.3-0.5 MPa and maintained for 2-4 hours.

6. The fibroblast compound solution for skin care according to claim 3, characterized in that: The further processing in step S3 includes adding an equal volume ratio of Antarctic krill oil to the oil.

7. The fibroblast complex solution for skin care according to claim 1, characterized in that: The preparation method of the Celastrus orbiculatus extract comprises the following steps: ① Dry the roots and stems of Celastrus orbiculatus and crush them to obtain Celastrus orbiculatus powder; ② adding ursolic acid with a mass concentration of 15-25% to the Celastrus orbiculatus powder at a material-liquid ratio of 1:(5-10) g / mL to obtain a premix; ③ Filter the premix to obtain a filtrate, and emulsify the filtrate at a speed of 3000-4000 r / min and a temperature of 25-30° C. for 40-50 minutes to obtain an emulsion; ④ Add tannic acid aqueous solution with a mass concentration of 2-5% to the emulsion, let it stand for 100-200 minutes and then dry it at a temperature of 60-80° C. to obtain the Celastrus orbiculatus extract.

8. The fibroblast complex solution for skin care according to claim 7, characterized in that: In step ④, the volume ratio of the emulsion to the tannic acid aqueous solution is 1:(1.5-3).

9. The fibroblast complex solution for skin care according to claim 1, characterized in that: The preparation method of the Potentilla anserine extract comprises the following steps: taking the whole plant of Potentilla anserine, drying it and then crushing it, mixing it with ethanol with a volume concentration of 70-80% at a solid-liquid ratio of 1:(3-5) g / mL, refluxing and extracting it for 60-100 minutes, collecting the extract, and concentrating it by reduced pressure evaporation to obtain the Potentilla anserine extract.

10. A method for preparing the fibroblast complex solution for skin care according to any one of claims 1 to 9, characterized in that: The steps include: (1) mixing fibroblast fluid, Rubescensine A and L-aspartic acid uniformly to prepare liquid A; (2) heating water to 60-70° C., adding Celastrus orbiculatus extract and Potentilla anserina extract, stirring evenly, cooling to 30-40° C., and then adding Cicadae confusa spore oil to prepare Liquid B; (3) Mix solution A and solution B, stir evenly, then add hydroxyethyl cellulose, stir and cool to 22-26° C. to obtain a fibroblast compound solution.

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