CN117138027A - Gel containing epidermal stem cell factor and application of gel in scar repair - Google Patents
Gel containing epidermal stem cell factor and application of gel in scar repair Download PDFInfo
- Publication number
- CN117138027A CN117138027A CN202311411723.6A CN202311411723A CN117138027A CN 117138027 A CN117138027 A CN 117138027A CN 202311411723 A CN202311411723 A CN 202311411723A CN 117138027 A CN117138027 A CN 117138027A
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- Prior art keywords
- gel
- epidermal
- oligopeptide
- stem cell
- factor
- Prior art date
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- Granted
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Classifications
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- A61K35/36—Skin; Hair; Nails; Sebaceous glands; Cerumen; Epidermis; Epithelial cells; Keratinocytes; Langerhans cells; Ectodermal cells
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- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/18—Growth factors; Growth regulators
- A61K38/1825—Fibroblast growth factor [FGF]
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- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/39—Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin, cold insoluble globulin [CIG]
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
- A61K47/32—Macromolecular compounds obtained by reactions only involving carbon-to-carbon unsaturated bonds, e.g. carbomers, poly(meth)acrylates, or polyvinyl pyrrolidone
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/0012—Galenical forms characterised by the site of application
- A61K9/0014—Skin, i.e. galenical aspects of topical compositions
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/06—Ointments; Bases therefor; Other semi-solid forms, e.g. creams, sticks, gels
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
- A61P17/02—Drugs for dermatological disorders for treating wounds, ulcers, burns, scars, keloids, or the like
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
- A61P37/02—Immunomodulators
Abstract
The invention discloses a gel containing an epidermal stem cell factor and application thereof in scar repair, and particularly relates to the field of cell engineering. Use of a gel comprising epidermal stem cell factor in scar repair. The gel comprises an epidermal cell supernatant extract, a fibroblast growth factor, carbomer, hyaluronic acid, nicotinamide, methylparaben, vitamins, collagen, oligopeptide-1 and oligopeptide-5. The gel containing the epidermal stem cell factor prepared by the invention can promote the regeneration of scar tissue: the epidermal cytokine gel can activate stem cells existing in the body and promote differentiation into cells with repair function, thereby promoting regeneration and repair of damaged tissues. This technique can more effectively promote the growth of new tissue and the healing of scars.
Description
Technical Field
The invention relates to the field of cell engineering, in particular to gel containing an epidermal stem cell factor and application thereof in scar repair.
Background
The scar is a local symptom which is left by fibrous tissue replacement repair and affects the appearance and the function, and the damage of physical, biological, chemical and other factors acts on the skin soft tissue of a human body, so that the skin soft tissue is seriously damaged and cannot be completely and normally repaired. The scars bring great physical and mental pain to the patient, especially scars left after burns, scalds and severe trauma. The period of scar hyperplasia is almost overwhelming for the patient for several years. The later atrophy period leads the patient to be completely non-planar and dysfunctional, and causes the patient to be extremely severely double-disturbance of the body and the heart.
Fibrous stem cell factor gels are currently used for repair and treatment of scar tissue. Such gels can promote the regeneration and repair process of the wound area by providing a suitable growth environment and active ingredients.
Fibroblasts are the main connective tissue cells existing in various tissues of the human body, and have important biological functions including synthesis of collagen, extracellular matrix, and the like. Stem cell factor is a special protein secreted by stem cells and can regulate cell proliferation, differentiation and function. By adding fibroblast and stem cell factor into gel, it can provide the growth factor and signal molecule needed by wound area, promote cell proliferation and tissue repair.
However, the existing fibroblast stem cell gel has the following defects: although fibroblast stem cell factor gel can promote repair of scar tissue, its efficacy may vary from individual to individual. For some severe scars, normal skin function may not be fully restored using this technique alone. Evidence of lack of long term validity: although fibroblast stem cell factor gels have shown some scar repair in laboratory and preliminary clinical studies, there is insufficient evidence for their long-term effectiveness. There is also a need for deep long-term follow-up studies to evaluate the persistence of their therapeutic effects. Lack of unified standards and specifications: in the use of fibroblast stem cell factor gel scar repairing technology, unified standards and specifications are lacking, so that great variability exists in clinical application of the technology. This presents some confusion to the clinician and patient.
Disclosure of Invention
Therefore, the invention provides a gel containing an epidermal stem cell factor and application thereof in scar repair, so as to solve the problems that the prior art is insufficient in curative effect and cannot be completely recovered.
In order to achieve the above object, the present invention provides the following technical solutions:
according to a first aspect of the present invention there is provided the use of a gel comprising epidermal stem cell factor in scar repair.
According to a second aspect of the present invention there is provided a gel comprising epidermal stem cell factor, comprising an epidermal cell supernatant extract, fibroblast growth factor, carbomer, hyaluronic acid, nicotinamide, methylparaben, vitamins, collagen, oligopeptide-1 and oligopeptide-5.
Further, the preparation method comprises 10-100ml of epidermal cell supernatant extract, 2-10ng/ml of fibroblast growth factor, 0.3-mg/ml of carbomer, 0.1-0.3mg/ml of hyaluronic acid, 0.2-0.4mg/ml of nicotinamide, 0.2-0.5mg/ml of methylparaben, 0.3-0.6mg/ml of vitamin, 0.2-0.6mg/ml of collagen, 0.1-0.3mg/ml of oligopeptide and 0.1-0.3mg/ml of oligopeptide.
Epidermal cell supernatant extract: can stimulate the synthesis and secretion of some macromolecules (such as hyaluronic acid, glycoprotein and the like) outside cells to moisten skin, so that the skin care cream can be used as an additive of cosmetics and used for facial plastic surgery, promote metabolism of human skin and reduce skin deformity; can also be used for treating skin injury, postoperative wound, decubital ulcer, canker sore, gangrene, dermatitis caused by radiotherapy, etc. It can accelerate healing of wounds and ulcer surfaces.
Fibroblast growth factor: can promote neovascularization and repair damaged endothelial cells.
Carbomer: has important applications such as thickening, suspending and the like, has simple process and good stability, and is widely applied to emulsion, cream and gel.
Hyaluronic acid: is an important component in skin and has the function of repairing epidermis tissue.
Nicotinamide: can promote biological oxidation process and tissue metabolism, and maintain normal tissue (especially skin) integrity.
Methylparaben: the broad-spectrum antibacterial activity can inhibit gram-negative bacteria and gram-positive bacteria, has a strong inhibition effect on saccharomycetes and mildew, and has the advantages of low use concentration, broad spectrum, high efficiency, safety, economy and long use period compared with the traditional antiseptic products (such as sodium benzoate). Is widely used for the corrosion prevention of cosmetics, medicines, foods and other industrial products.
Vitamin E: the vitamin E has the main functions of protecting skin, repairing damaged cell membranes, recovering normal functions of cells, promoting blood circulation and helping to fade stasis marks.
Collagen: can maintain the morphology and structure of skin and tissue organs, and is also an important raw material substance for repairing various damaged tissues.
Oligopeptide-1: can promote cell growth and increase cell activity, and repair damaged skin. Repairing collagen fibers and elastic fibers.
Oligopeptide-3: can induce stem cell differentiation, promote tissue repair, and repair basal layer fibroblast.
Banquet peptide 5: specifically stimulate epithelial cell metabolism, repair horny layer, increase fish thickness, and enhance collagen and elastin activity.
Furthermore, the epidermal cell supernatant extract is obtained by adding epidermal stem cells into an induction culture medium containing an induction drug to perform induction directional culture, performing moderate damage treatment, and finally collecting and purifying nutritional factors secreted by nutrition.
Further, the epidermal cell induction medicine comprises vitamin E, oligopeptide-3, fibroblast growth factor, trehalose, cysteine and rapamycin.
Further, the epidermal cell induction medicine comprises 0.01 mu L/ml vitamin E, 0.3 mu L/ml oligopeptide-3, 0.2 mu L/ml fibroblast growth factor, 0.2 mu L/ml trehalose, 0.1 mu L/ml cysteine and 0.5 mu L/ml rapamycin.
Vitamin E: vitamin E is an important antioxidant that helps protect cells and tissues from oxidative damage.
Oligopeptide-3: can induce stem cell differentiation, promote tissue repair, and repair basal layer fibroblast.
Fibroblast growth factor: can promote neovascularization and repair damaged endothelial cells.
Trehalose: can be used for cell regeneration.
Cysteine: cysteine can participate in various oxidation-reduction reactions in vivo and has a certain protection effect on cells.
Rapamycin: inducing autophagy and resisting aging.
According to a third aspect of the present invention, there is provided a method for preparing a gel comprising epidermal stem cell factor, the method comprising:
dissolving carbomer in an epidermal cell supernatant extract to obtain a carbomer solution;
sequentially adding a Xiang Kabo-mu solution into a fibroblast growth factor, hyaluronic acid, nicotinamide, methylparaben, vitamins, collagen, oligopeptide-1 and oligopeptide-5, and uniformly stirring to obtain a mixed solution;
and thirdly, placing the mixed solution at 23-28 ℃ for 40-60min to obtain gel solution.
The invention has the following advantages:
1. the gel containing the epidermal stem cell factor prepared by the invention can promote the regeneration of scar tissue: the epidermal cytokine gel can activate stem cells existing in the body and promote differentiation into cells with repair function, thereby promoting regeneration and repair of damaged tissues. This technique can more effectively promote the growth of new tissue and the healing of scars.
2. The gel containing the epidermal stem cell factor prepared by the invention has low risk: the epidermal cytokine gel scar repairing technology is a relatively safe treatment method.
3. The gel containing the epidermal stem cell factor prepared by the invention has multifunction: in addition to promoting scar repair, epidermal cytokine gels have other versatility. It can promote angiogenesis, anti-inflammatory effect, immunomodulation, etc., and can also help other problems in cell regeneration and tissue repair processes.
4. The gel containing the epidermal stem cell factor prepared by the invention can repair scars by utilizing the mixed liquid extracted by the epidermal cells to secrete various factors. Such as: EGF (epidermal growth factor), FGF (epidermal growth factor), VEGF (vascular endothelial growth factor), BFGF (neurotrophic factor), KGF (fibroblast growth factor).
5. The gel containing the epidermal stem cell factor prepared by the invention uses the cells with the next highest generation, generally the cells of the 6 th to 7 th generation, has more available amount per unit primary cell and reduced cost, and can be more efficiently used for clinical treatment research of scar repair.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below. It will be apparent to those of ordinary skill in the art that the drawings in the following description are exemplary only and that other implementations can be obtained from the extensions of the drawings provided without inventive effort.
The structures, proportions, sizes, etc. shown in the present specification are shown only for the purposes of illustration and description, and are not intended to limit the scope of the invention, which is defined by the claims, so that any structural modifications, proportional changes, or adjustments of size, which do not affect the efficacy of the invention or the objects achieved, should fall within the scope of the invention.
FIG. 1 is a graph showing the comparison of the induction of epidermal cells before and after the induction of epidermal cells according to example 1 of the present invention; wherein, before A-induction; b-after induction;
FIG. 2 is a Western blot detection chart of CD31, cutin 19 and collagen expression provided in example 1 of the present invention;
FIG. 3 is a graph showing the levels of the EGFR in the case of example 1 before and after induction;
FIG. 4 is a graph showing 1cm of the experimental example 2 of the present invention 2 Volunteers with scar use of a gel front-to-back comparison; wherein, A-before use; b, after using;
FIG. 5 is a 10cm chart of experimental example 2 of the invention 2 Volunteers with scar use of a gel front-to-back comparison; wherein, A-before use; and B, after the use.
Detailed Description
Other advantages and advantages of the present invention will become apparent to those skilled in the art from the following detailed description, which, by way of illustration, is to be read in connection with certain specific embodiments, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
Example 1
The present example provides an epidermal cell supernatant extract:
the seventh representative skin cells are spread on a six-hole plate and are cultured by using an alpha-MEM culture medium, the growth density reaches 80 percent, the cell growth state is good, and the cell activity rate is good>95%, the morphology accords with the morphology of the epidermal stem cells, and the cell surface molecular flow result accords with the molecular index related to the epidermal stem cells: the cell morphology of ci2B1, Q3B1, Q5B1, a6B4, ay35 and the like is observed to conform to the epidermic stem cell morphology under a microscope. Sucking out supernatant, washing with PBS buffer repeatedly for 3 times, sucking residual solution of PBS buffer after washing, slowly adding 2 ml/hole of induction culture medium (induction medicine: vitamin E0.01 μL/ml, oligopeptide-3.3 μL/ml, fibroblast growth factor 0.2 μL/ml, trehalose 0.2 μL/ml, cysteine 0.01 μL/ml, rapamycin 0.5 μL/ml) to the wall of the well plate, and placing the well plate into incubator (5% CO 2 The method comprises the steps of inducing epidermal cells for 72 hours to conduct directional differentiation, wherein a comparison chart before and after induction is shown in figure 1, treating the cells after induced directional differentiation by adopting an anoxic (8% oxygen+95% air, 37 ℃) method to cause moderate damage of the cells, collecting nutritional factors secreted by damaged nutrition, purifying (epidermal cell supernatant extract), and combining a mixture of ingredients for application.
Western blot detection of CD31, keratin 19, collagen expression is shown in FIG. 2.
The epidermal cell supernatant is collected and counted (using a countstar automatic cytometer), and then the results are subjected to Elisa detection of factors such as EGF (epidermal growth factor), FGF (epidermal growth factor), VEGF (vascular endothelial growth factor), BFGF (neurotrophic factor), KGF (fibroblast growth factor), and the like, and plotted. The results are shown in FIG. 3.
Example 2
The present example provides a gel containing epidermal stem cell factor:
10ml of epidermal cell supernatant extract, 2ng/ml of fibroblast growth factor, 0.3. 0.3mg/ml of carbomer, 0.1mg/ml of hyaluronic acid, 0.2mg/ml of nicotinamide, 0.2mg/ml of methylparaben, 0.3mg/ml of vitamin E, 0.2mg/ml of collagen, 0.1mg/ml of oligopeptide-1 and 0.1mg/ml of oligopeptide-5.
Example 3
The present example provides a gel containing epidermal stem cell factor:
50ml of epidermal cell supernatant extract, 5ng/ml of fibroblast growth factor, 0.3. 0.3mg/ml of carbomer, 0.2mg/ml of hyaluronic acid, 0.3mg/ml of nicotinamide, 0.23mg/ml of methylparaben, 0.5mg/ml of vitamin E, 0.4mg/ml of collagen, 0.2mg/ml of oligopeptide-1 and 0.2mg/ml of oligopeptide-5.
Example 4
The present example provides a gel containing epidermal stem cell factor:
the supernatant extract of the epidermal cells is 100ml, the growth factor of the fibroblast is 10ng/ml, the carbomer is 0.3mg/ml, the hyaluronic acid is 0.3mg/ml, the nicotinamide is 0.4mg/ml, the methylparaben is 0.5mg/ml, the vitamin E is 0.6mg/ml, the collagen is 0.6mg/ml, the oligopeptide-1.3 mg/ml and the oligopeptide-5.3 mg/ml.
Example 5
This example provides a method for preparing the epidermal stem cytokine-containing gel of examples 2-4:
1) Dissolving carbomer in the epidermal cell supernatant extract to obtain carbomer solution;
2) Sequentially adding Xiang Kabo m solution into fibroblast growth factor, hyaluronic acid, nicotinamide, methylparaben, vitamin E, collagen, oligopeptide-1 and oligopeptide-5, and stirring uniformly to obtain mixed solution;
3) And (3) placing the mixed solution at 23-28 ℃ for 40-60min to obtain a gel solution.
Experimental example 1
SD rat skin wound healing test SD rat skin defect model establishment and treatment. 30 male SD rats of 7-8 weeks old are selected, after the rats are anesthetized, the hairs of the backs of the rats are shaved, and a wound with a diameter of about 1.5cm is cut on the backs of the rats by using sterile ophthalmic scissors. SD rat models were randomly divided into 6 groups, example 1 group (6, wound site coated with scar gel prepared in example 2), example 2 group (6, wound site coated with scar gel prepared in example 3), example 3 group (6, wound site coated with scar gel prepared in example 3), example 4 group (6, wound site coated with scar gel prepared in example 4), and comparative example (6, wound site was sterilized with iodophor). Gel was applied 1 time a day in the morning and evening.
And (5) observing results. Wound healing was observed at 2d, 4d, 6d, 8d, 10d, 12d, 14d, respectively, and scar formation was observed. Wound healing rate (%) = (day 0 wound area-day n wound area)/day 0 wound area×100%. The experimental results are shown in the following table. The wound healing rate results are shown in table 1; scar formation is shown in table 2.
TABLE 1 wound healing Rate Table
TABLE 2 scar formation
As can be seen from the results in table 1, the wound healing rates of the examples 2 to 4 and the comparative example were significantly different from those of the example 1, and the examples were all effective in promoting wound healing, but the wound treated in the example 1 had a healing rate of more than 90% on day 6 and substantially complete healing on day 14, and the effect was more excellent.
Experimental example 2
Single wound area at 1cm was recruited 2 And 10cm 2 1 volunteer with scar on body, and the patient has good health status and no other systemic diseases, and signs informed consent before the test.
The body has a length of 1cm 2 Volunteers of scars, scars were smeared with scars gels containing epidermal stem cell factor prepared by the experimental group. The scar repairing gel is uniformly smeared on the scar every day, and the scar repairing gel is used once in the morning and at the evening, no other products are used in the morning and evening, and after the scar repairing gel is normally used for 10 days, the scar is obviously lightened and has a color similar to that of surrounding skin as shown in figure 4.
The body has a length of 10cm 2 Volunteers of scars, scars were smeared with scars gels containing epidermal stem cell factor prepared by the experimental group. The scar repairing gel is uniformly smeared on the scar every day, and the scar repairing gel is used once in the morning and at the evening, no other products are used in the morning and evening, and after the scar repairing gel is normally used for two months, as shown in figure 5, the scar is obviously lighter and has a color similar to that of surrounding skin.
It follows that the gel of the invention is able to promote scar repair with a lower risk. The gel of the invention has strong versatility and has anti-inflammatory and immunoregulatory effects. The invention utilizes the multi-factor secretion extraction mixed solution to repair the scar. The cells of the invention are used for a plurality of times, thereby reducing the cost. The invention can provide better help for scar repair research.
While the invention has been described in detail in the foregoing general description and specific examples, it will be apparent to those skilled in the art that modifications and improvements can be made thereto. Accordingly, such modifications or improvements may be made without departing from the spirit of the invention and are intended to be within the scope of the invention as claimed.
Claims (7)
1. Use of a gel comprising epidermal stem cell factor in scar repair.
2. A gel containing epidermal stem cell factor, which comprises an epidermal cell supernatant extract, a fibroblast growth factor, carbomer, hyaluronic acid, nicotinamide, methylparaben, vitamins, collagen, oligopeptide-1 and oligopeptide-5.
3. The gel of claim 2, comprising 10-100ml of epidermal cell supernatant extract, 2-10ng/ml of fibroblast growth factor, 0.3-mg/ml of carbomer, 0.1-0.3mg/ml of hyaluronic acid, 0.2-0.4mg/ml of nicotinamide, 0.2-0.5mg/ml of methylparaben, 0.3-0.6mg/ml of vitamins, 0.2-0.6mg/ml of collagen, 0.1-0.3mg/ml of oligopeptide and 0.1-0.3mg/ml of oligopeptide.
4. The gel containing epidermal stem cell factor of claim 2, wherein the epidermal cell supernatant extract is obtained by adding epidermal stem cells into an induction medium containing an induction drug to perform induction directional culture, performing moderate injury treatment, and finally collecting and purifying nutrient factors secreted by nutrition.
5. The gel of claim 4, wherein the epidermal cell-inducing agent comprises vitamin E, oligopeptide-3, fibroblast growth factor, trehalose, cysteine, rapamycin.
6. The gel of claim 5, wherein the epidermal cell-inducing agent comprises vitamin E0.01. Mu.L/ml, oligopeptide-3.3. Mu.L/ml, fibroblast growth factor 0.2. Mu.L/ml, trehalose 0.2. Mu.L/ml, cysteine 0.1. Mu.L/ml, rapamycin 0.5. Mu.L/ml.
7. A method of preparing an epidermal stem cell factor-containing gel, comprising:
dissolving carbomer in an epidermal cell supernatant extract to obtain a carbomer solution;
sequentially adding a Xiang Kabo-mu solution into a fibroblast growth factor, hyaluronic acid, nicotinamide, methylparaben, vitamins, collagen, oligopeptide-1 and oligopeptide-5, and uniformly stirring to obtain a mixed solution;
and thirdly, placing the mixed solution at 23-28 ℃ for 40-60min to obtain gel solution.
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| JP2020137518A (en) * | 2019-02-25 | 2020-09-03 | 株式会社細胞応用技術研究所 | Additive |
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| CN108125903A (en) * | 2018-01-19 | 2018-06-08 | 深圳光彩生命工程技术有限公司 | A kind of novel stem cells composite factor anti-aging cosmetics |
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