CN112641714A - Preparation method of essence containing autologous adipose-derived stem cell cytokines - Google Patents
Preparation method of essence containing autologous adipose-derived stem cell cytokines Download PDFInfo
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- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
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Abstract
The invention discloses a preparation method of essence containing autologous adipose-derived stem cell factors. According to the invention, the autologous adipose-derived stem cell factors are added, and are rich in active factor components such as epidermal growth factors, vascular endothelial growth factors and stem cell factors, so that the factors can promote the growth and proliferation of stem cells in vivo, promote fibroblasts in the skin and the like to secrete more collagen, can obviously relieve the aging of the skin, and are added with the newly prepared high-lubrication moisturizing silicone wax and ascorbyl glucoside, so that the effects of efficiently moisturizing, whitening and lightening spots are achieved.
Description
Technical Field
The invention belongs to the technical field of cosmetic preparation, and relates to a preparation method of essence containing autologous adipose-derived stem cell cytokines.
Background
In recent years, beauty treatment by using living cells becomes a popular hot spot in the market, particularly the use of stem cells, and living cells can differentiate and grow in the skin by directly injecting on wrinkles, scars and scars, so that the beauty treatment agent plays a role in filling and repairing the skin. Most of the applied living cells are from homologous sources and allogeneic, and the immune rejection is generated when the living cells enter the autoimmune system in the body, and the generated cosmetic effect is low. The adipose-derived stem cell is a novel adult stem cell which is discovered in recent years, has strong proliferation capacity, can maintain stable growth and proliferation activity in vitro culture, has multiple differentiation potentials, and is also applied to cosmetic treatment.
However, the beauty effect is realized by simply using the adipose-derived stem cell extracting solution through an injection mode, the efficiency of directionally differentiating the adipose-derived stem cells into the adipose cells and the fibroblasts is low, the effect is slow, a professional needs to use medical equipment in a specified place through an injection method, the crowds of the professional are limited, and in addition, the product containing the autologous adipose-derived stem cell factors and having the effects of whitening and moisturizing is few, so that the problem can be solved by developing the essence containing the autologous adipose-derived stem cell factors and having the effects of moisturizing and whitening.
Disclosure of Invention
The invention aims to provide a preparation method of essence containing autologous adipose-derived stem cell factors, which is characterized in that autologous adipose-derived stem cell factors are added, and the autologous adipose-derived stem cell factors are rich in active factor components such as epidermal growth factors, vascular endothelial growth factors, stem cell factors and the like, can promote the growth and proliferation of stem cells in vivo, promote fibroblasts in the skin and the like to secrete more collagen, can obviously relieve the aging of the skin, and are added with newly-prepared high-lubrication moisturizing silicone wax and ascorbic acid glucoside, so that the effects of efficiently moisturizing, whitening and lightening spots are exerted.
The purpose of the invention can be realized by the following technical scheme:
a preparation method of essence containing autologous adipose-derived stem cell cytokines comprises the following steps:
step A1, selecting autologous adipose-derived stem cells from a cell bank, washing and centrifuging, discarding supernatant, re-suspending the cells discarded with autologous adipose-derived stem cell culture solution without phenol red, inoculating the cells in a culture bottle, culturing for 18-24 hours, collecting cell supernatant, and sieving to obtain autologous adipose-derived stem cell cytokines;
step A2, stirring and mixing sodium hyaluronate, collagen, denatured ethanol, high-lubrication moisturizing silicone wax and hydroxyethyl piperazine ethanesulfonic acid in a stirring tank at 45-50 ℃ and at 250-300rpm for 30-60 minutes to prepare a mixture A;
step A3, adding the mixture A prepared in the step A2, octyldodecanol, ascorbic acid glucoside, polyacrylamide dimethyl ammonium taurate and deionized water into an emulsifying pot, and emulsifying at 70-80 ℃ for 10-15 minutes to prepare a mixture B;
and step A4, cooling the temperature of the emulsification pot in the step A3 to room temperature, adding the autologous adipose-derived stem cell cytokine prepared in the step A1 into the compound B, and stirring and mixing at 150-160rpm for 20-30 minutes to prepare the essence containing the autologous adipose-derived stem cell cytokine.
Further, the sodium hyaluronate, the collagen, the denatured ethanol, the high-lubrication moisturizing silicone wax and the hydroxyethylpiperazine ethanesulfonic acid in the step A2 are 20-25 parts by weight of sodium hyaluronate, 5-8 parts by weight of collagen, 1-3 parts by weight of denatured ethanol, 10-15 parts by weight of the high-lubrication moisturizing silicone wax and 4-6 parts by weight of hydroxyethylpiperazine ethanesulfonic acid.
Furthermore, the weight parts of the octyl dodecanol, the ascorbic acid glucoside, the polyacrylamide dimethyl taurate and the deionized water in the step A3 are 4-6 parts of octyl dodecanol, 8-10 parts of ascorbic acid glucoside, 2-4 parts of polyacrylamide dimethyl taurate and 6-8 parts of deionized water.
Further, the weight parts of the autologous adipose-derived stem cell cytokines in the step A4 are 4-6 parts.
Further, the high-lubrication moisturizing silicone wax is prepared by the following steps:
step S1, adding hydrogen silicone oil and 1-octadecene into a three-neck flask, heating to 80-95 ℃, slowly adding a catalyst chloroplatinic acid-isopropanol solution, heating to the temperature required by the reaction of 120 ℃ and 140 ℃, reacting for 3-5 hours, heating to 150 ℃ after the reaction is finished, keeping the vacuum at-0.1 MPa, and removing the 1-octadecene through reduced pressure distillation to obtain dialkyl terminated polysiloxane;
step S2, adding dialkyl end-capped polysiloxane and cyclotetrasiloxane prepared in the step S1 into a three-neck flask, adding catalyst sulfonated cation resin, controlling the reaction temperature to be 80-90 ℃, the stirring speed to be 200-210rpm, reacting for 5-6 hours, stopping heating and stirring after the reaction is finished, cooling to room temperature, removing cation exchange resin by suction filtration, heating to 110-120 ℃, keeping the vacuum-0.1 MPa, removing cyclotetrasiloxane by reduced pressure distillation, adding sodium bicarbonate to adjust the pH value to be 7-8, filtering under reduced pressure, and treating a filter cake for 2-4 days by active carbon to obtain a dialkyl end-capped side chain hydrogen-containing organosilicon intermediate;
and step S3, adding the dialkyl-terminated side chain hydrogen-containing organosilicon intermediate and 1-octadecene prepared in the step S2 into a three-neck flask, heating to 90-95 ℃, slowly adding a catalyst chloroplatinic acid-isopropanol solution, heating to 140-150 ℃, reacting for 6-8 hours, heating to 170 ℃ after the reaction is finished, keeping the vacuum-0.1 MPa, and removing 1-octadecene through reduced pressure distillation to obtain the high-lubrication moisture-retention organosilicon wax.
Further, the dosage ratio of the hydrogen-terminated silicone oil and the 1-octadecene in the step S1 is 6-8 g: 30-40mL of hydrogen-terminated silicone oil with the molecular weight of 2000, 1-2mL of chloroplatinic acid-isopropanol solution and 0.8mL of chloroplatinic acid added into 100mL of isopropanol.
Furthermore, the dosage of the cyclotetrasiloxane in the step S2 is 4-6g, and the dosage of the sulfonated cationic resin is 5% -10% of the mass of the cyclotetrasiloxane.
Further, the dosage ratio of the 1-octadecene and the chloroplatinic acid-isopropanol solution in the step S3 is 3-5 g: 1-2mL, chloroplatinic acid-isopropanol solution was prepared by adding 1mL chloroplatinic acid per 100mL isopropanol.
The invention has the beneficial effects that: by adding the autologous adipose-derived stem cell factors, the autologous adipose-derived stem cell factors are rich in active factor components such as epidermal growth factors, vascular endothelial growth factors, stem cell factors and the like, can promote the growth and proliferation of stem cells in vivo, promote fibroblasts in the skin and the like to secrete more collagen, can obviously relieve the aging of the skin, and are added with newly-prepared high-lubrication moisturizing silicone wax and ascorbic acid glucoside to exert the effects of efficiently moisturizing, whitening and lightening spots.
Detailed Description
The technical solutions of the present invention will be described clearly and completely with reference to the following embodiments, and it should be understood that the described embodiments are only a part of the embodiments of the present invention, and not all of the 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
The high-lubrication moisturizing organosilicone wax is prepared by the following steps:
step S1, adding 6g of hydrogen-terminated silicone oil and 30mL of 1-octadecene into a three-neck flask, heating to 80 ℃, slowly adding 1mL of chloroplatinic acid-isopropanol solution serving as a catalyst, heating to 120 ℃ required by reaction, reacting for 3 hours, heating to 150 ℃ after the reaction is finished, keeping the vacuum of-0.1 MPa, and removing 1-octadecene through reduced pressure distillation to obtain dialkyl terminated polysiloxane;
step S2, adding the dialkyl-terminated polysiloxane prepared in the step S1 and 4g of cyclotetrasiloxane into a three-neck flask, adding 0.2g of catalyst sulfonated cation resin, controlling the reaction temperature to be 80 ℃, the stirring rate to be 200rpm, reacting for 5 hours, stopping heating and stirring after the reaction is finished, cooling to room temperature, removing cation exchange resin by suction filtration, heating to 110 ℃, keeping the vacuum at-0.1 MPa, removing cyclotetrasiloxane by reduced pressure distillation, adding sodium bicarbonate to adjust the pH value to be 7, filtering under reduced pressure, and treating a filter cake with activated carbon for 2 days to obtain a dialkyl-terminated side chain hydrogen-containing organosilicon intermediate;
and step S3, adding the dialkyl-terminated side chain hydrogen-containing organosilicon intermediate prepared in the step S2 and 3g of 1-octadecene into a three-neck flask, heating to 90 ℃, slowly adding 1mL of chloroplatinic acid-isopropanol solution serving as a catalyst, heating to 140 ℃, reacting for 6 hours, heating to 170 ℃ after the reaction is finished, keeping the vacuum at minus 0.1MPa, and removing 1-octadecene through reduced pressure distillation to prepare the high-lubrication moisture-retention organosilicon wax.
A preparation method of essence containing autologous adipose-derived stem cell cytokines comprises the following steps:
step A1, selecting autologous adipose-derived stem cells from a cell bank, washing and centrifuging, discarding supernatant, re-suspending the cells discarded with autologous adipose-derived stem cell culture solution without phenol red, inoculating the cells in a culture bottle, culturing for 18 hours, collecting cell supernatant, and sieving to obtain autologous adipose-derived stem cell cytokines;
step A2, stirring and mixing 20 parts of sodium hyaluronate, 5 parts of collagen, 1 part of denatured ethanol, 10 parts of high-lubrication moisturizing silicone wax and 4 parts of hydroxyethyl piperazine ethanesulfonic acid in a stirring tank at 45 ℃ and 250rpm for 30 minutes to prepare a mixture A;
step A3, adding the mixture A prepared in the step A2, 4 parts of octyldodecanol, 8 parts of ascorbyl glucoside, 2 parts of ammonium polyacryloyldimethyl taurate and 6 parts of deionized water into an emulsifying pot, and emulsifying at 80 ℃ for 10 minutes to prepare a mixture B;
and step A4, cooling the temperature of the emulsifying pot in the step A3 to room temperature, adding 4 parts of the autologous adipose-derived stem cell cytokines prepared in the step A1 into the compound B, and stirring and mixing at 150rpm for 30 minutes to prepare the essence containing the autologous adipose-derived stem cell cytokines.
Example 2
The high-lubrication moisturizing organosilicone wax is prepared by the following steps:
step S1, adding 7g of hydrogen-terminated silicone oil and 30mL of 1-octadecene into a three-neck flask, heating to 80 ℃, slowly adding 2mL of chloroplatinic acid-isopropanol solution serving as a catalyst, heating to 120 ℃ required by reaction, reacting for 3 hours, heating to 150 ℃ after the reaction is finished, keeping the vacuum of-0.1 MPa, and removing 1-octadecene through reduced pressure distillation to obtain dialkyl terminated polysiloxane;
step S2, adding the dialkyl-terminated polysiloxane prepared in the step S1 and 5g of cyclotetrasiloxane into a three-neck flask, adding 0.25g of catalyst sulfonated cation resin, controlling the reaction temperature to be 80 ℃, the stirring rate to be 200rpm, reacting for 5 hours, stopping heating and stirring after the reaction is finished, cooling to room temperature, removing cation exchange resin by suction filtration, heating to 110 ℃, keeping the vacuum at-0.1 MPa, removing cyclotetrasiloxane by reduced pressure distillation, adding sodium bicarbonate to adjust the pH value to be 7, filtering under reduced pressure, and treating a filter cake with activated carbon for 2 days to obtain a dialkyl-terminated side chain hydrogen-containing organosilicon intermediate;
and step S3, adding the dialkyl-terminated side chain hydrogen-containing organosilicon intermediate prepared in the step S2 and 4g of 1-octadecene into a three-neck flask, heating to 90 ℃, slowly adding 2mL of chloroplatinic acid-isopropanol solution serving as a catalyst, heating to 140 ℃, reacting for 6 hours, heating to 170 ℃ after the reaction is finished, keeping the vacuum at minus 0.1MPa, and removing 1-octadecene through reduced pressure distillation to obtain the high-lubrication and moisture-retention organosilicon wax.
A preparation method of essence containing autologous adipose-derived stem cell cytokines comprises the following steps:
step A1, selecting autologous adipose-derived stem cells from a cell bank, washing and centrifuging, discarding supernatant, re-suspending the cells discarded with autologous adipose-derived stem cell culture solution without phenol red, inoculating the cells in a culture bottle, culturing for 24 hours, collecting cell supernatant, and sieving to obtain autologous adipose-derived stem cell cytokines;
step A2, stirring and mixing 22 parts of sodium hyaluronate, 6 parts of collagen, 2 parts of denatured ethanol, 12 parts of high-lubrication moisturizing silicone wax and 5 parts of hydroxyethyl piperazine ethanesulfonic acid in a stirring tank at 45 ℃ and 250rpm for 30 minutes to prepare a mixture A;
step A3, adding the mixture A prepared in the step A2, 5 parts of octyldodecanol, 9 parts of ascorbyl glucoside, 3 parts of ammonium polyacryloyldimethyl taurate and 7 parts of deionized water into an emulsifying pot, and emulsifying at 80 ℃ for 10 minutes to prepare a mixture B;
and step A4, cooling the temperature of the emulsifying pot in the step A3 to room temperature, adding 5 parts of the autologous adipose-derived stem cell cytokines prepared in the step A1 into the compound B, and stirring and mixing at 150rpm for 30 minutes to prepare the essence containing the autologous adipose-derived stem cell cytokines.
Example 3
The high-lubrication moisturizing organosilicone wax is prepared by the following steps:
step S1, adding 8g of hydrogen-terminated silicone oil and 40mL of 1-octadecene into a three-neck flask, heating to 80 ℃, slowly adding 1mL of chloroplatinic acid-isopropanol solution serving as a catalyst, heating to 120 ℃ required by reaction, reacting for 3 hours, heating to 150 ℃ after the reaction is finished, keeping the vacuum of-0.1 MPa, and removing 1-octadecene through reduced pressure distillation to obtain dialkyl terminated polysiloxane;
step S2, adding the dialkyl-terminated polysiloxane prepared in the step S1 and 6g of cyclotetrasiloxane into a three-neck flask, adding 0.3g of catalyst sulfonated cation resin, controlling the reaction temperature to be 80 ℃, the stirring rate to be 200rpm, reacting for 5 hours, stopping heating and stirring after the reaction is finished, cooling to room temperature, removing cation exchange resin by suction filtration, heating to 110 ℃, keeping the vacuum at-0.1 MPa, removing cyclotetrasiloxane by reduced pressure distillation, adding sodium bicarbonate to adjust the pH value to be 7, filtering under reduced pressure, and treating a filter cake with activated carbon for 2 days to obtain a dialkyl-terminated side chain hydrogen-containing organosilicon intermediate;
and step S3, adding the dialkyl-terminated side chain hydrogen-containing organosilicon intermediate prepared in the step S2 and 5g of 1-octadecene into a three-neck flask, heating to 90 ℃, slowly adding 1mL of chloroplatinic acid-isopropanol solution serving as a catalyst, heating to 140 ℃, reacting for 6 hours, heating to 170 ℃ after the reaction is finished, keeping the vacuum at minus 0.1MPa, and removing 1-octadecene through reduced pressure distillation to prepare the high-lubrication moisture-retention organosilicon wax.
A preparation method of essence containing autologous adipose-derived stem cell cytokines comprises the following steps:
step A1, selecting autologous adipose-derived stem cells from a cell bank, washing and centrifuging, discarding supernatant, re-suspending the cells discarded with autologous adipose-derived stem cell culture solution without phenol red, inoculating the cells in a culture bottle, culturing for 24 hours, collecting cell supernatant, and sieving to obtain autologous adipose-derived stem cell cytokines;
step A2, stirring and mixing 25 parts of sodium hyaluronate, 8 parts of collagen, 3 parts of denatured ethanol, 15 parts of high-lubricity moisturizing silicone wax and 6 parts of hydroxyethyl piperazine ethanesulfonic acid in a stirring tank at 45 ℃ and 250rpm for 30 minutes to prepare a mixture A;
step A3, adding the mixture A prepared in the step A2, 6 parts of octyldodecanol, 10 parts of ascorbyl glucoside, 4 parts of polyacrylamide dimethyl ammonium taurate and 8 parts of deionized water into an emulsifying pot, and emulsifying at 80 ℃ for 10 minutes to prepare a mixture B;
and step A4, cooling the temperature of the emulsifying pot in the step A3 to room temperature, adding 6 parts of the autologous adipose-derived stem cell cytokines prepared in the step A1 into the compound B, and stirring and mixing the mixture at 150rpm for 30 minutes to prepare the essence containing the autologous adipose-derived stem cell cytokines.
Comparative example 1
And (4) common essence.
The essence prepared in examples 1 to 3 and comparative example 1 were subjected to efficacy test, 100 female subjects with significant wrinkles and spots on their faces over 40 years old were selected and evenly divided into 5 groups, and the essence was applied every morning and evening to observe the efficacy, and the test data are shown in table 1:
TABLE 1
As can be seen from the table, the essence containing autologous adipose-derived stem cell cytokines prepared in examples 1 to 3 has excellent effects of repairing wrinkles and lightening spots, and also has the functions of moisturizing and whitening.
The preferred embodiments of the invention disclosed above are intended to be illustrative only. The preferred embodiments are not intended to be exhaustive or to limit the invention to the precise embodiments disclosed. Obviously, many modifications and variations are possible in light of the above teaching. The embodiments were chosen and described in order to best explain the principles of the invention and the practical application, to thereby enable others skilled in the art to best utilize the invention. The invention is limited only by the claims and their full scope and equivalents.
Claims (8)
1. The preparation method of the essence containing the autologous adipose-derived stem cell cytokines is characterized by comprising the following steps of:
step A1, selecting autologous adipose-derived stem cells from a cell bank, washing and centrifuging, discarding supernatant, re-suspending the cells discarded with autologous adipose-derived stem cell culture solution without phenol red, inoculating the cells in a culture bottle, culturing for 18-24 hours, collecting cell supernatant, and sieving to obtain autologous adipose-derived stem cell cytokines;
step A2, stirring and mixing sodium hyaluronate, collagen, denatured ethanol, high-lubrication moisturizing silicone wax and hydroxyethyl piperazine ethanesulfonic acid in a stirring tank at 45-50 ℃ and at 250-300rpm for 30-60 minutes to prepare a mixture A;
step A3, adding the mixture A prepared in the step A2, octyldodecanol, ascorbic acid glucoside, polyacrylamide dimethyl ammonium taurate and deionized water into an emulsifying pot, and emulsifying at 70-80 ℃ for 10-15 minutes to prepare a mixture B;
and step A4, cooling the temperature of the emulsification pot in the step A3 to room temperature, adding the autologous adipose-derived stem cell cytokine prepared in the step A1 into the compound B, and stirring and mixing at 150-160rpm for 20-30 minutes to prepare the essence containing the autologous adipose-derived stem cell cytokine.
2. The method for preparing essence containing autologous adipose-derived stem cell cytokines according to claim 1, wherein the method comprises the following steps: the sodium hyaluronate, the collagen, the denatured ethanol, the high-lubrication moisturizing silicone wax and the hydroxyethylpiperazine ethanesulfonic acid in the step A2 are 20-25 parts by weight of sodium hyaluronate, 5-8 parts by weight of collagen, 1-3 parts by weight of denatured ethanol, 10-15 parts by weight of high-lubrication moisturizing silicone wax and 4-6 parts by weight of hydroxyethylpiperazine ethanesulfonic acid.
3. The method for preparing essence containing autologous adipose-derived stem cell cytokines according to claim 1, wherein the method comprises the following steps: the weight parts of the octyl dodecanol, the ascorbic acid glucoside, the polyacrylamide dimethyl taurate and the deionized water in the step A3 are 4-6 parts of octyl dodecanol, 8-10 parts of ascorbic acid glucoside, 2-4 parts of polyacrylamide dimethyl taurate and 6-8 parts of deionized water.
4. The method for preparing essence containing autologous adipose-derived stem cell cytokines according to claim 1, wherein the method comprises the following steps: the weight part of the autologous adipose-derived stem cell cytokine in the step A4 is 4-6 parts.
5. The method for preparing essence containing autologous adipose-derived stem cell cytokines according to claim 1, wherein the method comprises the following steps: the high-lubrication moisturizing silicone wax is prepared by the following steps:
step S1, adding hydrogen silicone oil and 1-octadecene into a three-neck flask, heating to 80-95 ℃, slowly adding a catalyst chloroplatinic acid-isopropanol solution, heating to the temperature required by the reaction of 120 ℃ and 140 ℃, reacting for 3-5 hours, heating to 150 ℃ after the reaction is finished, keeping the vacuum at-0.1 MPa, and removing the 1-octadecene through reduced pressure distillation to obtain dialkyl terminated polysiloxane;
step S2, adding dialkyl end-capped polysiloxane and cyclotetrasiloxane prepared in the step S1 into a three-neck flask, adding catalyst sulfonated cation resin, controlling the reaction temperature to be 80-90 ℃, the stirring speed to be 200-210rpm, reacting for 5-6 hours, stopping heating and stirring after the reaction is finished, cooling to room temperature, removing cation exchange resin by suction filtration, heating to 110-120 ℃, keeping the vacuum-0.1 MPa, removing cyclotetrasiloxane by reduced pressure distillation, adding sodium bicarbonate to adjust the pH value to be 7-8, filtering under reduced pressure, and treating a filter cake for 2-4 days by active carbon to obtain a dialkyl end-capped side chain hydrogen-containing organosilicon intermediate;
and step S3, adding the dialkyl-terminated side chain hydrogen-containing organosilicon intermediate and 1-octadecene prepared in the step S2 into a three-neck flask, heating to 90-95 ℃, slowly adding a catalyst chloroplatinic acid-isopropanol solution, heating to 140-150 ℃, reacting for 6-8 hours, heating to 170 ℃ after the reaction is finished, keeping the vacuum-0.1 MPa, and removing 1-octadecene through reduced pressure distillation to obtain the high-lubrication moisture-retention organosilicon wax.
6. The method for preparing essence containing autologous adipose-derived stem cell cytokines according to claim 5, wherein the method comprises the following steps: the dosage ratio of the hydrogen-terminated silicone oil to the 1-octadecene in the step S1 is 6-8 g: 30-40mL of hydrogen-terminated silicone oil with the molecular weight of 2000, 1-2mL of chloroplatinic acid-isopropanol solution and 0.8mL of chloroplatinic acid added into 100mL of isopropanol.
7. The method for preparing essence containing autologous adipose-derived stem cell cytokines according to claim 5, wherein the method comprises the following steps: the dosage of the cyclotetrasiloxane in the step S2 is 4-6g, and the dosage of the sulfonated cationic resin is 5-10% of the mass of the cyclotetrasiloxane.
8. The method for preparing essence containing autologous adipose-derived stem cell cytokines according to claim 5, wherein the method comprises the following steps: the dosage ratio of the 1-octadecene and the chloroplatinic acid-isopropanol solution in the step S3 is 3-5 g: 1-2mL, chloroplatinic acid-isopropanol solution was prepared by adding 1mL chloroplatinic acid per 100mL isopropanol.
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| Publication number | Priority date | Publication date | Assignee | Title |
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