CN112675068A - Preparation method of face cream of autologous adipose-derived stem cell cytokine extracting solution - Google Patents
Preparation method of face cream of autologous adipose-derived stem cell cytokine extracting solution Download PDFInfo
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- 239000006071 cream Substances 0.000 title claims abstract description 43
- 210000000130 stem cell Anatomy 0.000 title claims abstract description 42
- 102000004127 Cytokines Human genes 0.000 title claims abstract description 30
- 108090000695 Cytokines Proteins 0.000 title claims abstract description 30
- 238000002360 preparation method Methods 0.000 title claims description 17
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims abstract description 32
- 239000008367 deionised water Substances 0.000 claims abstract description 23
- 229910021641 deionized water Inorganic materials 0.000 claims abstract description 23
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 23
- BXWNKGSJHAJOGX-UHFFFAOYSA-N hexadecan-1-ol Chemical compound CCCCCCCCCCCCCCCCO BXWNKGSJHAJOGX-UHFFFAOYSA-N 0.000 claims abstract description 22
- PRAKJMSDJKAYCZ-UHFFFAOYSA-N squalane Chemical compound CC(C)CCCC(C)CCCC(C)CCCCC(C)CCCC(C)CCCC(C)C PRAKJMSDJKAYCZ-UHFFFAOYSA-N 0.000 claims abstract description 22
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 claims abstract description 20
- 230000003020 moisturizing effect Effects 0.000 claims abstract description 15
- 239000000284 extract Substances 0.000 claims abstract description 13
- NIXOWILDQLNWCW-UHFFFAOYSA-N Acrylic acid Chemical compound OC(=O)C=C NIXOWILDQLNWCW-UHFFFAOYSA-N 0.000 claims abstract description 11
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 11
- 229920002385 Sodium hyaluronate Polymers 0.000 claims abstract description 11
- 229920002125 Sokalan® Polymers 0.000 claims abstract description 11
- CBTVGIZVANVGBH-UHFFFAOYSA-N aminomethyl propanol Chemical compound CC(C)(N)CO CBTVGIZVANVGBH-UHFFFAOYSA-N 0.000 claims abstract description 11
- 229960001631 carbomer Drugs 0.000 claims abstract description 11
- 229960000541 cetyl alcohol Drugs 0.000 claims abstract description 11
- 239000004200 microcrystalline wax Substances 0.000 claims abstract description 11
- 235000019808 microcrystalline wax Nutrition 0.000 claims abstract description 11
- JXTPJDDICSTXJX-UHFFFAOYSA-N n-Triacontane Natural products CCCCCCCCCCCCCCCCCCCCCCCCCCCCCC JXTPJDDICSTXJX-UHFFFAOYSA-N 0.000 claims abstract description 11
- 239000004006 olive oil Substances 0.000 claims abstract description 11
- 235000008390 olive oil Nutrition 0.000 claims abstract description 11
- 229940010747 sodium hyaluronate Drugs 0.000 claims abstract description 11
- YWIVKILSMZOHHF-QJZPQSOGSA-N sodium;(2s,3s,4s,5r,6r)-6-[(2s,3r,4r,5s,6r)-3-acetamido-2-[(2s,3s,4r,5r,6r)-6-[(2r,3r,4r,5s,6r)-3-acetamido-2,5-dihydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-2-carboxy-4,5-dihydroxyoxan-3-yl]oxy-5-hydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-3,4,5-trihydroxyoxane-2- Chemical compound [Na+].CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@H](O[C@H]2[C@@H]([C@@H](O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@H](O3)C(O)=O)O)[C@H](O)[C@@H](CO)O2)NC(C)=O)[C@@H](C(O)=O)O1 YWIVKILSMZOHHF-QJZPQSOGSA-N 0.000 claims abstract description 11
- 229940032094 squalane Drugs 0.000 claims abstract description 11
- JSOVGYMVTPPEND-UHFFFAOYSA-N 16-methylheptadecyl 2,2-dimethylpropanoate Chemical compound CC(C)CCCCCCCCCCCCCCCOC(=O)C(C)(C)C JSOVGYMVTPPEND-UHFFFAOYSA-N 0.000 claims abstract description 10
- ZZZCUOFIHGPKAK-UHFFFAOYSA-N D-erythro-ascorbic acid Natural products OCC1OC(=O)C(O)=C1O ZZZCUOFIHGPKAK-UHFFFAOYSA-N 0.000 claims abstract description 10
- 229930003268 Vitamin C Natural products 0.000 claims abstract description 10
- 235000019154 vitamin C Nutrition 0.000 claims abstract description 10
- 239000011718 vitamin C Substances 0.000 claims abstract description 10
- 239000002994 raw material Substances 0.000 claims abstract description 7
- 238000000034 method Methods 0.000 claims abstract description 4
- 239000003795 chemical substances by application Substances 0.000 claims abstract 5
- 238000003756 stirring Methods 0.000 claims description 54
- 239000000243 solution Substances 0.000 claims description 51
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 45
- 229920002101 Chitin Polymers 0.000 claims description 40
- 238000006243 chemical reaction Methods 0.000 claims description 19
- 239000007788 liquid Substances 0.000 claims description 19
- 239000000203 mixture Substances 0.000 claims description 19
- ROSDSFDQCJNGOL-UHFFFAOYSA-N Dimethylamine Chemical compound CNC ROSDSFDQCJNGOL-UHFFFAOYSA-N 0.000 claims description 16
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 15
- 239000000706 filtrate Substances 0.000 claims description 15
- CEAZRRDELHUEMR-URQXQFDESA-N Gentamicin Chemical compound O1[C@H](C(C)NC)CC[C@@H](N)[C@H]1O[C@H]1[C@H](O)[C@@H](O[C@@H]2[C@@H]([C@@H](NC)[C@@](C)(O)CO2)O)[C@H](N)C[C@@H]1N CEAZRRDELHUEMR-URQXQFDESA-N 0.000 claims description 14
- 229930182566 Gentamicin Natural products 0.000 claims description 14
- 229960002518 gentamicin Drugs 0.000 claims description 14
- JHJLBTNAGRQEKS-UHFFFAOYSA-M sodium bromide Chemical compound [Na+].[Br-] JHJLBTNAGRQEKS-UHFFFAOYSA-M 0.000 claims description 14
- ZORQXIQZAOLNGE-UHFFFAOYSA-N 1,1-difluorocyclohexane Chemical compound FC1(F)CCCCC1 ZORQXIQZAOLNGE-UHFFFAOYSA-N 0.000 claims description 10
- 238000001914 filtration Methods 0.000 claims description 10
- 239000001593 sorbitan monooleate Substances 0.000 claims description 10
- 229940035049 sorbitan monooleate Drugs 0.000 claims description 10
- 235000011069 sorbitan monooleate Nutrition 0.000 claims description 10
- 238000001816 cooling Methods 0.000 claims description 9
- 239000000758 substrate Substances 0.000 claims description 9
- QAOWNCQODCNURD-UHFFFAOYSA-N sulfuric acid Substances OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 claims description 9
- OEDMOCYNWLHUDP-UHFFFAOYSA-N bromomethanol Chemical compound OCBr OEDMOCYNWLHUDP-UHFFFAOYSA-N 0.000 claims description 8
- FOCAUTSVDIKZOP-UHFFFAOYSA-N chloroacetic acid Chemical compound OC(=O)CCl FOCAUTSVDIKZOP-UHFFFAOYSA-N 0.000 claims description 8
- 229940106681 chloroacetic acid Drugs 0.000 claims description 8
- 239000002245 particle Substances 0.000 claims description 8
- NNJVILVZKWQKPM-UHFFFAOYSA-N Lidocaine Chemical compound CCN(CC)CC(=O)NC1=C(C)C=CC=C1C NNJVILVZKWQKPM-UHFFFAOYSA-N 0.000 claims description 7
- 229960004194 lidocaine Drugs 0.000 claims description 7
- 239000005708 Sodium hypochlorite Substances 0.000 claims description 5
- 238000005119 centrifugation Methods 0.000 claims description 5
- 210000001268 chyle Anatomy 0.000 claims description 5
- SUKJFIGYRHOWBL-UHFFFAOYSA-N sodium hypochlorite Chemical compound [Na+].Cl[O-] SUKJFIGYRHOWBL-UHFFFAOYSA-N 0.000 claims description 5
- 230000008961 swelling Effects 0.000 claims description 5
- 238000010257 thawing Methods 0.000 claims description 5
- 239000008187 granular material Substances 0.000 claims description 4
- 238000000967 suction filtration Methods 0.000 claims description 4
- 239000002504 physiological saline solution Substances 0.000 claims description 2
- WNWHHMBRJJOGFJ-UHFFFAOYSA-N 16-methylheptadecan-1-ol Chemical compound CC(C)CCCCCCCCCCCCCCCO WNWHHMBRJJOGFJ-UHFFFAOYSA-N 0.000 claims 2
- AKKLAJYCGVIWBS-UHFFFAOYSA-N O=[N].CC1(C)CCCC(C)(C)N1 Chemical compound O=[N].CC1(C)CCCC(C)(C)N1 AKKLAJYCGVIWBS-UHFFFAOYSA-N 0.000 claims 2
- 235000011187 glycerol Nutrition 0.000 claims 2
- 239000012530 fluid Substances 0.000 claims 1
- 239000000543 intermediate Substances 0.000 claims 1
- 239000003921 oil Substances 0.000 claims 1
- 235000019198 oils Nutrition 0.000 claims 1
- 229950010765 pivalate Drugs 0.000 claims 1
- IUGYQRQAERSCNH-UHFFFAOYSA-N pivalic acid Chemical compound CC(C)(C)C(O)=O IUGYQRQAERSCNH-UHFFFAOYSA-N 0.000 claims 1
- 238000013517 stratification Methods 0.000 claims 1
- 239000003906 humectant Substances 0.000 abstract description 20
- KIUKXJAPPMFGSW-DNGZLQJQSA-N (2S,3S,4S,5R,6R)-6-[(2S,3R,4R,5S,6R)-3-Acetamido-2-[(2S,3S,4R,5R,6R)-6-[(2R,3R,4R,5S,6R)-3-acetamido-2,5-dihydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-2-carboxy-4,5-dihydroxyoxan-3-yl]oxy-5-hydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-3,4,5-trihydroxyoxane-2-carboxylic acid Chemical compound CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@H](O[C@H]2[C@@H]([C@@H](O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@H](O3)C(O)=O)O)[C@H](O)[C@@H](CO)O2)NC(C)=O)[C@@H](C(O)=O)O1 KIUKXJAPPMFGSW-DNGZLQJQSA-N 0.000 abstract description 4
- 229920002674 hyaluronan Polymers 0.000 abstract description 4
- 229960003160 hyaluronic acid Drugs 0.000 abstract description 4
- 150000004676 glycans Chemical class 0.000 abstract description 2
- 239000004615 ingredient Substances 0.000 abstract description 2
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- 239000005017 polysaccharide Substances 0.000 abstract description 2
- JNYAEWCLZODPBN-JGWLITMVSA-N (2r,3r,4s)-2-[(1r)-1,2-dihydroxyethyl]oxolane-3,4-diol Chemical compound OC[C@@H](O)[C@H]1OC[C@H](O)[C@H]1O JNYAEWCLZODPBN-JGWLITMVSA-N 0.000 abstract 1
- RKMGAJGJIURJSJ-UHFFFAOYSA-N 2,2,6,6-tetramethylpiperidine Chemical compound CC1(C)CCCC(C)(C)N1 RKMGAJGJIURJSJ-UHFFFAOYSA-N 0.000 description 10
- MWUXSHHQAYIFBG-UHFFFAOYSA-N Nitric oxide Chemical compound O=[N] MWUXSHHQAYIFBG-UHFFFAOYSA-N 0.000 description 8
- 230000000694 effects Effects 0.000 description 8
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- 210000004027 cell Anatomy 0.000 description 4
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- Cosmetics (AREA)
Abstract
本发明公开了一种自体脂肪源干细胞细胞因子提取液的面霜制备方法，该面霜包括如下重量份原料：自体脂肪源干细胞细胞因子提取液1‑3份、卡波姆0.5‑0.8份、甘油5‑10份、氨甲基丙醇0.2‑0.5份、维生素C1‑2份、乙酰化六肽0.5‑1份、去离子水50‑80份、保湿剂1‑3份、角鲨烷1‑5份、橄榄油10‑15份、鲸蜡醇3‑5份、微晶蜡2‑5份、异硬脂醇新戊酸酯0.5‑1份、透明质酸钠0.05‑0.2份、失水山梨醇单油酸酯0.5‑1份；该保湿剂是一种聚合多糖,与皮肤有很好的亲合能力,其结构与人体皮肤中存在的天然保湿成分透明质酸接近,因此对皮肤具有良好的保湿性、润湿性。The invention discloses a method for preparing a face cream of an autologous adipose-derived stem cell cytokine extract. The face cream comprises the following raw materials in parts by weight: 1-3 parts of an autologous adipose-derived stem cell cytokine extract, 0.5-0.8 parts of carbomer, and 5 parts of glycerol. -10 parts, aminomethyl propanol 0.2-0.5 parts, vitamin C 1-2 parts, acetylated hexapeptide 0.5-1 part, deionized water 50-80 parts, humectant 1-3 parts, squalane 1-5 parts parts, 10-15 parts of olive oil, 3-5 parts of cetyl alcohol, 2-5 parts of microcrystalline wax, 0.5-1 part of isostearyl pivalate, 0.05-0.2 part of sodium hyaluronate, sorbitan 0.5-1 part of alcohol monooleate; the moisturizing agent is a polymeric polysaccharide, has good affinity with the skin, and its structure is close to the natural moisturizing ingredient hyaluronic acid existing in human skin, so it has good skin moisturizing and moisturizing properties.
Description
Technical Field
The invention relates to the technical field of facial cream preparation, and particularly relates to a preparation method of facial cream of an autologous adipose-derived stem cell factor extracting solution.
Background
Autologous stem cells are "seed cells". The animal body realizes the cell renewal through the division of the autologous stem cells, thereby ensuring the continuous growth and development of the animal body. When a disease, an organ is struck or injured, autologous stem cells respond first time, promoting differentiation into the exclusive cells needed by the body, thereby providing sufficient numbers of exclusive cells to aid in repair and healing. Has wide application in the treatment of various diseases such as nervous system diseases, immune system diseases and the like. And europe has applied this technology to anti-aging and early prevention of disease.
The existing face cream has more using functions, but the effect is general and the face cream effect is single, after part of face cream is used for 1-2h, the moisturizing effect cannot be achieved, and after the face cream is used for a long time, the skin has no obvious improvement effect, so that the market popularization is not facilitated.
Disclosure of Invention
The invention aims to provide a method for preparing face cream of an autologous adipose-derived stem cell cytokine extracting solution.
The technical problems to be solved by the invention are as follows:
the existing face cream has more using functions, but the effect is general and the face cream effect is single, after part of face cream is used for 1-2h, the moisturizing effect cannot be achieved, and after the face cream is used for a long time, the skin has no obvious improvement effect, so that the market popularization is not facilitated.
The purpose of the invention can be realized by the following technical scheme:
a preparation method of a face cream of an autologous adipose-derived stem cell cytokine extracting solution comprises the following raw materials in parts by weight: 1-3 parts of autologous fat-derived stem cell cytokine extracting solution, 0.5-0.8 part of carbomer, 5-10 parts of glycerol, 0.2-0.5 part of aminomethyl propanol, 0-2 parts of vitamin C1, 0.5-1 part of acetylated hexapeptide, 50-80 parts of deionized water, 1-3 parts of humectant, 1-5 parts of squalane, 10-15 parts of olive oil, 3-5 parts of cetyl alcohol, 2-5 parts of microcrystalline wax, 0.5-1 part of isostearyl pivalate, 0.05-0.2 part of sodium hyaluronate and 0.5-1 part of sorbitan monooleate;
the preparation method of the face cream specifically comprises the following steps:
step S1: adding the autologous adipose-derived stem cell cytokine extracting solution, carbomer, glycerol, aminomethyl propanol, vitamin C, acetylated hexapeptide and deionized water into a stirring kettle, and stirring for 2-3h at the rotation speed of 300-;
step S2: adding squalane, olive oil, cetyl alcohol, microcrystalline wax, isostearyl pivalate and sodium hyaluronate into a stirring kettle, and continuously stirring for 1-1.5 hours at the rotation speed of 800-;
step S3: adding the first mixture and the second mixture into a stirring kettle, stirring for 3-5min at a rotation speed of 3000r/min and a temperature of 75-80 ℃, cooling to 60 ℃, adding the humectant and the sorbitan monooleate, continuing stirring for 20-30min, and cooling to room temperature to obtain the cream.
Further, the autologous adipose-derived stem cell factor extracting solution is prepared by the following steps:
step A1: adding normal saline into autologous fat liquid, standing at 5-10 deg.C for 3-5min until layering, discharging bottom liquid, centrifuging the rest liquid at 400r/min for 2-3min, and collecting bottom swelling liquid to obtain fat particles;
step A2: the fat particle chyle is ground and then mixed with normal saline and gentamicin, four layers of layers are layered after centrifugation is carried out for 5min at the temperature of 3-5 ℃ and the rotating speed of 2500r/min, top layer grease and third layer normal saline are removed, and a substrate, gentamicin and lidocaine are uniformly mixed to prepare the autologous adipose-derived stem cell factor extracting solution.
Further, the dosage volume ratio of the fat particles, the physiological saline and the gentamicin in the step A2 is 25:23:2, the dosage of the gentamicin is 4U per 15mL of the substrate, and the dosage volume ratio of the lidocaine and the substrate is 1: 5.
Further, the humectant is prepared by the following steps:
step B1: dispersing chitin in a sodium hydroxide solution, stirring for 1-1.5h under the condition that the rotation speed is 200-300r/min, freezing for 10-15h under the condition that the temperature is 8-10 ℃, thawing and filtering to obtain pretreated chitin;
step B2: dispersing pretreated chitin in deionized water, adding sodium bromide and 2,2,6, 6-tetramethylpiperidine nitric oxide, stirring at the rotation speed of 150-200r/min until the pretreated chitin is completely dissolved, adding a sodium hypochlorite solution with the pH value of 10, reacting at the temperature of 0 ℃ and keeping the pH value of the reaction solution at 10, filtering to remove filtrate after reacting for 2-3h, adding ethanol into the filtrate and standing for 10-15min, and filtering to remove the filtrate to obtain oxidized chitin;
step B3: adding a dimethylamine solution and bromomethanol into a reaction kettle, stirring for 2-3h under the condition that the rotation speed is 200-300r/min to prepare an intermediate 1, adding the intermediate 1 and the oxidized chitin prepared in the step B2 into the reaction kettle, stirring and adding concentrated sulfuric acid under the conditions that the rotation speed is 150-200r/min and the temperature is 70-80 ℃ to react for 3-5h to prepare an intermediate 2, mixing chloroacetic acid and sodium hydroxide until the pH value is 7, adding deionized water and the intermediate 2, and reacting at the temperature of 50-55 ℃ to prepare the humectant.
The reaction process is as follows:
further, the volume fraction of the sodium hydroxide solution in the step B1 is 40%, the mass ratio of the pretreated chitin, sodium bromide and 2,2,6, 6-tetramethylpiperidine oxynitride in the step B2 is 1:0.125:0.0125, the molar ratio of the dimethylamine solution and bromomethanol in the step B3 is 1:1, the mass ratio of the intermediate 1, oxidized chitin and concentrated sulfuric acid is 1g:3g:5mL, the mass fraction of concentrated sulfuric acid is 95%, and the mass ratio of chloroacetic acid, sodium hydroxide, deionized water and the intermediate 2 is 0.1mol:0.1mol:10mL:0.1 mol.
The invention has the beneficial effects that: the invention extracts the autologous adipose-derived stem cells in the process of preparing the face cream of the autologous adipose-derived stem cell cytokine extracting solution, the autologous adipose-derived stem cell extract can form epidermal cells, bone, cartilage cells and stem cells and can be differentiated to form myocardial cells, nerve cells, vascular endothelial cells and blood cells so as to regenerate tissues to achieve the effect of repairing skin, and simultaneously a humectant is prepared, the humectant takes chitin as a raw material, firstly the chitin is treated to prepare pretreated chitin, the pretreated chitin is oxidized to prepare oxidized chitin, a dimethylamine solution and bromomethanol are reacted to prepare an intermediate 1, the intermediate 1 and the oxidized chitin are subjected to oxidation reaction to prepare an intermediate 2, the intermediate 2 is treated to prepare the humectant which is a polymeric polysaccharide, has good affinity with skin, no allo-allergy, safety, no toxicity or irritation, and good biocompatibility. Meanwhile, the structure of the hyaluronic acid moisturizing cream is close to that of hyaluronic acid which is a natural moisturizing ingredient existing in human skin, so that the hyaluronic acid moisturizing cream has good moisturizing and moistening effects on the skin.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Example 1
A preparation method of a face cream of an autologous adipose-derived stem cell cytokine extracting solution comprises the following raw materials in parts by weight: 1 part of autologous fat-derived stem cell cytokine extracting solution, 0.5 part of carbomer, 5 parts of glycerol, 0.2 part of aminomethyl propanol, 1 parts of vitamin C, 0.5 part of acetylated hexapeptide, 50 parts of deionized water, 1 part of humectant, 1 part of squalane, 10 parts of olive oil, 3 parts of cetyl alcohol, 2 parts of microcrystalline wax, 0.5 part of isostearyl pivalate, 0.05 part of sodium hyaluronate and 0.5 part of sorbitan monooleate;
the preparation method of the face cream specifically comprises the following steps:
step S1: adding the autologous adipose-derived stem cell cytokine extracting solution, carbomer, glycerol, aminomethyl propanol, vitamin C, acetylated hexapeptide and deionized water into a stirring kettle, and stirring for 2 hours at the rotating speed of 300r/min and the temperature of 50 ℃ to prepare a first mixture;
step S2: adding squalane, olive oil, cetyl alcohol, microcrystalline wax, isostearyl pivalate and sodium hyaluronate into a stirring kettle, and continuously stirring for 1h at the rotation speed of 800r/min and the temperature of 75 ℃ to prepare a second mixture;
step S3: adding the first mixture and the second mixture into a stirring kettle, stirring for 3min at a rotation speed of 3000r/min and a temperature of 75 ℃, cooling to a temperature of 60 ℃, adding a humectant and sorbitan monooleate, continuing stirring for 20min, and cooling to room temperature to obtain the cream.
The autologous adipose-derived stem cell factor extracting solution is prepared by the following steps:
step A1: adding normal saline into autologous fat liquid, standing at 5 deg.C for 3min for layering, discharging bottom liquid, centrifuging the rest liquid at 400r/min for 2min, and collecting bottom swelling liquid to obtain fat granules;
step A2: the fat particle chyle is crushed and then mixed with normal saline and gentamicin, after centrifugation is carried out for 5min at the temperature of 3 ℃ and the rotating speed of 2500r/min, four layers of layering are generated, top layer grease and third layer normal saline are removed, and a substrate, gentamicin and lidocaine are uniformly mixed to prepare the autologous adipose-derived stem cell cytokine extracting solution.
The humectant is prepared by the following steps:
step B1: dispersing chitin in a sodium hydroxide solution, stirring for 1h at a rotation speed of 200r/min, freezing for 10h at a temperature of-8 ℃, thawing, and performing suction filtration to obtain pretreated chitin;
step B2: dispersing pretreated chitin in deionized water, adding sodium bromide and 2,2,6, 6-tetramethylpiperidine nitric oxide, stirring at the rotation speed of 150r/min until the pretreated chitin is completely dissolved, adding a sodium hypochlorite solution with the pH value of 10, reacting at the temperature of 0 ℃ while keeping the pH value of the reaction solution at 10, filtering to remove a filtrate after reacting for 2 hours, adding ethanol into the filtrate and standing for 10 minutes, and filtering to remove the filtrate to prepare oxidized chitin;
step B3: adding a dimethylamine solution and bromomethanol into a reaction kettle, stirring for 2 hours at the rotating speed of 200r/min to prepare an intermediate 1, adding the intermediate 1 and the oxidized chitin prepared in the step B2 into the reaction kettle, stirring and adding concentrated sulfuric acid at the rotating speed of 150r/min and the temperature of 70 ℃ to react for 3 hours to prepare an intermediate 2, mixing chloroacetic acid and sodium hydroxide until the pH value is 7, adding deionized water and the intermediate 2, and reacting at the temperature of 50 ℃ to prepare the humectant.
Example 2
A preparation method of a face cream of an autologous adipose-derived stem cell cytokine extracting solution comprises the following raw materials in parts by weight: 2 parts of autologous fat-derived stem cell cytokine extracting solution, 0.6 part of carbomer, 8 parts of glycerol, 0.3 part of aminomethyl propanol, 1.5 parts of vitamin C, 0.8 part of acetylated hexapeptide, 60 parts of deionized water, 2 parts of humectant, 3 parts of squalane, 13 parts of olive oil, 4 parts of cetyl alcohol, 3 parts of microcrystalline wax, 0.8 part of isostearyl pivalate, 0.1 part of sodium hyaluronate and 0.8 part of sorbitan monooleate;
the preparation method of the face cream specifically comprises the following steps:
step S1: adding the autologous adipose-derived stem cell cytokine extracting solution, carbomer, glycerol, aminomethyl propanol, vitamin C, acetylated hexapeptide and deionized water into a stirring kettle, and stirring for 2 hours at the rotating speed of 300r/min and the temperature of 60 ℃ to prepare a first mixture;
step S2: adding squalane, olive oil, cetyl alcohol, microcrystalline wax, isostearyl pivalate and sodium hyaluronate into a stirring kettle, and continuously stirring for 1.5 hours at the rotation speed of 1000r/min and the temperature of 75 ℃ to prepare a second mixture;
step S3: adding the first mixture and the second mixture into a stirring kettle, stirring at a rotation speed of 3000r/min and a temperature of 75 ℃ for 5min, cooling to a temperature of 60 ℃, adding a humectant and sorbitan monooleate, continuing stirring for 20min, and cooling to room temperature to obtain the cream.
The autologous adipose-derived stem cell factor extracting solution is prepared by the following steps:
step A1: adding normal saline into autologous fat liquid, standing at 10 deg.C for 3min for layering, discharging bottom liquid, centrifuging the rest liquid at 400r/min for 3min, and collecting bottom swelling liquid to obtain fat granules;
step A2: the fat particle chyle is crushed and then mixed with normal saline and gentamicin, after centrifugation is carried out for 5min at the temperature of 3 ℃ and the rotating speed of 2500r/min, four layers of layering are generated, top layer grease and third layer normal saline are removed, and a substrate, gentamicin and lidocaine are uniformly mixed to prepare the autologous adipose-derived stem cell cytokine extracting solution.
The humectant is prepared by the following steps:
step B1: dispersing chitin in a sodium hydroxide solution, stirring for 1h at the rotation speed of 300r/min, freezing for 10h at the temperature of minus 10 ℃, thawing, and performing suction filtration to obtain pretreated chitin;
step B2: dispersing pretreated chitin in deionized water, adding sodium bromide and 2,2,6, 6-tetramethylpiperidine nitric oxide, stirring at the rotation speed of 200r/min until the pretreated chitin is completely dissolved, adding a sodium hypochlorite solution with the pH value of 10, reacting at the temperature of 0 ℃ while keeping the pH value of the reaction solution at 10, filtering to remove a filtrate after reacting for 2 hours, adding ethanol into the filtrate and standing for 15min, and filtering to remove the filtrate to obtain oxidized chitin;
step B3: adding a dimethylamine solution and bromomethanol into a reaction kettle, stirring for 3 hours at the rotating speed of 200r/min to prepare an intermediate 1, adding the intermediate 1 and the oxidized chitin prepared in the step B2 into the reaction kettle, stirring and adding concentrated sulfuric acid at the rotating speed of 150r/min and the temperature of 80 ℃ to react for 3 hours to prepare an intermediate 2, mixing chloroacetic acid and sodium hydroxide until the pH value is 7, adding deionized water and the intermediate 2, and reacting at the temperature of 55 ℃ to prepare the humectant.
Example 3
A preparation method of a face cream of an autologous adipose-derived stem cell cytokine extracting solution comprises the following raw materials in parts by weight: 3 parts of autologous fat-derived stem cell cytokine extracting solution, 0.8 part of carbomer, 10 parts of glycerol, 0.5 part of aminomethyl propanol, 2 parts of vitamin C, 1 part of acetylated hexapeptide, 80 parts of deionized water, 3 parts of humectant, 5 parts of squalane, 15 parts of olive oil, 5 parts of cetyl alcohol, 5 parts of microcrystalline wax, 1 part of isostearyl pivalate, 0.2 part of sodium hyaluronate and 1 part of sorbitan monooleate;
the preparation method of the face cream specifically comprises the following steps:
step S1: adding the autologous adipose-derived stem cell cytokine extracting solution, carbomer, glycerol, aminomethyl propanol, vitamin C, acetylated hexapeptide and deionized water into a stirring kettle, and stirring for 3 hours at the rotating speed of 500r/min and the temperature of 60 ℃ to prepare a first mixture;
step S2: adding squalane, olive oil, cetyl alcohol, microcrystalline wax, isostearyl pivalate and sodium hyaluronate into a stirring kettle, and continuously stirring for 1.5 hours at the rotation speed of 1000r/min and the temperature of 80 ℃ to prepare a second mixture;
step S3: adding the first mixture and the second mixture into a stirring kettle, stirring at a rotation speed of 3000r/min and a temperature of 80 ℃ for 5min, cooling to a temperature of 60 ℃, adding a humectant and sorbitan monooleate, continuing stirring for 30min, and cooling to room temperature to obtain the cream.
The autologous adipose-derived stem cell factor extracting solution is prepared by the following steps:
step A1: adding normal saline into autologous fat liquid, standing at 10 deg.C for 5min for layering, discharging bottom liquid, centrifuging the rest liquid at 400r/min for 3min, and collecting bottom swelling liquid to obtain fat granules;
step A2: the fat particle chyle is crushed and then mixed with normal saline and gentamicin, after centrifugation is carried out for 5min at the temperature of 5 ℃ and the rotating speed of 2500r/min, four layers of layers are layered, top layer grease and third layer normal saline are removed, and a substrate, gentamicin and lidocaine are uniformly mixed to prepare the autologous adipose-derived stem cell cytokine extracting solution.
The humectant is prepared by the following steps:
step B1: dispersing chitin in sodium hydroxide solution, stirring for 1.5h at the rotation speed of 300r/min, freezing for 15h at the temperature of-10 ℃, thawing, and performing suction filtration to obtain pretreated chitin;
step B2: dispersing pretreated chitin in deionized water, adding sodium bromide and 2,2,6, 6-tetramethylpiperidine nitric oxide, stirring at the rotation speed of 200r/min until the pretreated chitin is completely dissolved, adding a sodium hypochlorite solution with the pH value of 10, reacting at the temperature of 0 ℃ while keeping the pH value of the reaction solution at 10, filtering to remove a filtrate after reacting for 3 hours, adding ethanol into the filtrate and standing for 15min, and filtering to remove the filtrate to obtain oxidized chitin;
step B3: adding a dimethylamine solution and bromomethanol into a reaction kettle, stirring for 3 hours at the rotating speed of 300r/min to prepare an intermediate 1, adding the intermediate 1 and the oxidized chitin prepared in the step B2 into the reaction kettle, stirring and adding concentrated sulfuric acid at the rotating speed of 200r/min and the temperature of 80 ℃ to react for 5 hours to prepare an intermediate 2, mixing chloroacetic acid and sodium hydroxide until the pH value is 7, adding deionized water and the intermediate 2, and reacting at the temperature of 55 ℃ to prepare the humectant.
Comparative example
The comparative example is a common facial cream on the market.
200 males and females aged 18 to 25 were randomly selected and divided into four groups, and the first group to the third group were applied with the face creams prepared in examples 1 to 3, respectively, and the fourth group was applied with the face cream prepared in comparative example, and used after cleansing every morning and evening, and the skin moisture rising ratio was compared for 30 days of continuous use, and the results are shown in the following table 1.
TABLE 1
| Example 1 | Example 2 | Example 3 | Comparative example | |
| Skin moisture rising ratio (%) | 26.31 | 25.43 | 25.68 | 11.29 |
As is apparent from Table 1 above, the skin moisture rising ratio of the creams obtained in examples 1 to 3 was 25.43 to 26.31, while the skin moisture rising ratio of the cream obtained in the comparative example was 11.29, indicating that the present invention has a very good moisturizing effect.
The foregoing is merely exemplary and illustrative of the principles of the present invention and various modifications, additions and substitutions of the specific embodiments described herein may be made by those skilled in the art without departing from the principles of the present invention or exceeding the scope of the claims set forth herein.
Claims (5)
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| CN113262195A (en) * | 2021-06-02 | 2021-08-17 | 上海南滨江细胞生物科技有限公司 | Face cream containing stem cell exosomes and preparation method thereof |
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