CN110812525A - Aseptic collagen liquid of adipose-derived stem cell exosome and preparation method of application - Google Patents
Aseptic collagen liquid of adipose-derived stem cell exosome and preparation method of application Download PDFInfo
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L15/00—Chemical aspects of, or use of materials for, bandages, dressings or absorbent pads
- A61L15/16—Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons
- A61L15/22—Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons containing macromolecular materials
- A61L15/32—Proteins, polypeptides; Degradation products or derivatives thereof, e.g. albumin, collagen, fibrin, gelatin
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/64—Proteins; Peptides; Derivatives or degradation products thereof
- A61K8/65—Collagen; Gelatin; Keratin; Derivatives or degradation products thereof
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- A—HUMAN NECESSITIES
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- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/98—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution of animal origin
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- A61L15/40—Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons containing ingredients of undetermined constitution or reaction products thereof, e.g. plant or animal extracts
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- A61L15/00—Chemical aspects of, or use of materials for, bandages, dressings or absorbent pads
- A61L15/16—Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons
- A61L15/42—Use of materials characterised by their function or physical properties
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
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Abstract
The invention discloses sterile collagen liquid of an adipose-derived stem cell exosome, which consists of the adipose-derived stem cell exosome and sterile collagen liquid, wherein each milliliter of the sterile collagen liquid contains 0.5-0.8mg of exosome. The preparation method comprises the following steps: mixing the adipose stem cell exosome and the sterile collagen liquid in a sterile environment, uniformly stirring, and storing at a low temperature. The invention also discloses a preparation method of the adipose-derived stem cell exosome by applying the sterile collagen liquid, which is characterized in that the sterile collagen liquid of the adipose-derived stem cell exosome is pressed between two non-woven fabrics under the aseptic condition, and then the non-woven fabrics are put into an aluminum foil bag to be sealed and stored at low temperature. The invention prepares the aseptic collagen liquid of the adipose-derived stem cell exosome by combining the adipose-derived stem cell exosome with the aseptic collagen liquid, can fully play the roles of the adipose-derived stem cell exosome and the aseptic collagen liquid, has better treatment effect on skin injury repair and wound repair, and can shorten the wound healing time.
Description
Technical Field
The invention relates to the technical field of sterile collagen liquid and application preparation, in particular to sterile collagen liquid of adipose-derived stem cell exosomes and a preparation method of the application.
Background
The collagen is fibrous extracellular protein formed by twisting three peptide chains into a spiral shape, accounts for 25 to 30 percent of the protein of the whole body, and has unique biological activity. The collagen is rich in amino acids such as glycine, proline, hydroxyproline and the like which are necessary for human bodies, is the most important component in extracellular matrix, has strong biological activity and biological function, can participate in the migration, differentiation and proliferation of cells, enables the bone tendon, the cartilage and the skin to have certain mechanical strength, and has wide application in the medical and sanitary fields of burn, wound, eye corneal diseases, cosmetology, hard tissue repair, wound hemostasis and the like due to weak antigenicity and good biocompatibility.
Exosomes are tiny vesicles secreted by most cells, with lipid bilayer membrane structures, approximately 30-150nm in diameter. Exosomes contain cell-specific proteins, lipids and nucleic acids, can be transmitted to other cells as signal molecules to change the functions of other cells, and play an important role in immune antigen presentation, tumor growth and migration, and repair of tissue damage. Adipose-derived stem cells are recently isolated from adipose tissues as a kind of stem cells having a multi-differentiation potential. Mainly recovers the repairing function of tissue cells, promotes the regeneration of cells, recovers the young face, simultaneously fully improves the body function, effectively improves diseases such as sub-health, premature senility and the like, and really and effectively resists aging from inside to outside.
At present, effective research on the combination of the adipose-derived stem cell exosomes and the sterile collagen liquid is less, so that the benefits of the combination of the adipose-derived stem cell exosomes and the sterile collagen liquid cannot be reflected, and the function exertion of the adipose-derived stem cell exosomes and the sterile collagen liquid product is limited to a certain extent.
Disclosure of Invention
In view of the above, the present invention provides a sterile collagen solution for adipose stem cell exosomes and a preparation method for application thereof.
In order to solve the technical problem, the invention discloses sterile collagen liquid of an adipose-derived stem cell exosome, which consists of the adipose-derived stem cell exosome and the sterile collagen liquid, wherein each milliliter of the sterile collagen liquid contains 0.5-0.8mg of the exosome.
Further, the sterile collagen liquid is fish skin collagen liquid.
The invention discloses a second technical scheme, which is a preparation method of aseptic collagen liquid of adipose-derived stem cell exosomes, wherein the adipose-derived stem cell exosomes and the aseptic collagen liquid are mixed under an aseptic environment, each milliliter of the aseptic collagen liquid contains 0.5-0.8mg of exosomes, and the aseptic collagen liquid of the adipose-derived stem cell exosomes is obtained by uniformly stirring and storing at low temperature.
Further, the sterile collagen liquid is fish skin collagen liquid.
The invention discloses a third technical scheme that a preparation method of aseptic collagen liquid application of adipose-derived stem cell exosomes specifically comprises the following steps:
step 1, under the environment with aseptic process control, two pieces of rectangular non-woven fabrics with the same size and specification are taken, and the aseptic collagen liquid of the prepared adipose-derived stem cell exosome is coated between one piece of non-woven fabric;
step 2, coating a layer of viscose on the periphery of the non-woven fabric coated with the aseptic collagen liquid of the adipose-derived stem cell exosomes, and then bonding the two non-woven fabrics together to enable the aseptic collagen liquid of the adipose-derived stem cell exosomes to be pressed between the two non-woven fabrics, so that the aseptic collagen liquid is obtained and applied;
and 3, sticking the sterile collagen liquid obtained in the step 2 in an aluminum foil bag, sealing the aluminum foil bag, and storing the aluminum foil bag at a low temperature.
Further, the coating thickness of the sterile collagen liquid of the adipose-derived stem cell exosomes in the step 1 is 2-3 mm.
Further, the thickness of the viscose in the step 2 is 0.5-1 mm.
The fourth technical scheme disclosed by the invention is that the prepared sterile collagen liquid of the adipose-derived stem cell exosome is applied to the preparation of skin injury repair products, wound repair products, medical beauty products or cosmetics.
The invention can obtain the following technical effects:
the aseptic collagen liquid of the adipose-derived stem cell exosome is prepared by combining the adipose-derived stem cell exosome and the aseptic collagen liquid, can fully exert the functions of the adipose-derived stem cell exosome and the aseptic collagen liquid, has better treatment effects on skin injury repair and wound repair, and can shorten the wound healing time.
The preparation method of the aseptic collagen liquid application of the adipose-derived stem cell exosome carries out the preparation of the application under the aseptic condition, reduces the subsequent sterilization steps, reduces the cost and is beneficial to keeping the intact bioactivity of collagen.
Detailed Description
The following embodiments are described in detail with reference to the accompanying drawings, so that how to implement the technical features of the present invention to solve the technical problems and achieve the technical effects can be fully understood and implemented.
The invention discloses sterile collagen liquid of an adipose-derived stem cell exosome, which consists of the adipose-derived stem cell exosome and sterile collagen liquid, wherein each milliliter of the sterile collagen liquid contains 0.5-0.8mg of exosome.
Wherein the aseptic collagen liquid is fish skin collagen liquid, and the processing method of the fish skin collagen liquid is the prior art.
The adipose-derived stem cell exosome is extracted by the existing method.
The invention also discloses a preparation method of the aseptic collagen liquid of the adipose-derived stem cell exosome, which comprises the steps of mixing the adipose-derived stem cell exosome and the aseptic collagen liquid in an aseptic environment, wherein each milliliter of the aseptic collagen liquid contains 0.5-0.8mg of exosome, uniformly stirring, and storing at low temperature to obtain the adipose-derived stem cell exosome.
The aseptic collagen liquid of the adipose-derived stem cell exosome is prepared by combining the adipose-derived stem cell exosome and the aseptic collagen liquid, can fully exert the functions of the adipose-derived stem cell exosome and the aseptic collagen liquid, has better treatment effects on skin injury repair and wound repair, and can shorten the wound healing time.
The invention also discloses a preparation method of the aseptic collagen liquid application of the adipose-derived stem cell exosome, which specifically comprises the following steps:
step 1, under the environment with aseptic process control, taking two rectangular non-woven fabrics with the same size and specification, and coating the prepared aseptic collagen liquid of the adipose-derived stem cell exosome in the middle of one non-woven fabric, wherein the coating thickness of the aseptic collagen liquid of the adipose-derived stem cell exosome is 2-3 mm;
step 2, coating a layer of viscose on the periphery of the non-woven fabric coated with the aseptic collagen liquid of the adipose-derived stem cell exosomes, wherein the thickness of the viscose is 0.5-1mm, and then bonding the two non-woven fabrics together to ensure that the aseptic collagen liquid of the adipose-derived stem cell exosomes is pressed between the two non-woven fabrics to obtain the aseptic collagen liquid application;
and 3, sticking the sterile collagen liquid obtained in the step 2 in an aluminum foil bag, sealing the aluminum foil bag, and storing the aluminum foil bag at a low temperature.
The preparation method of the aseptic collagen liquid application of the adipose-derived stem cell exosome carries out the preparation of the application under the aseptic condition, reduces the subsequent sterilization steps, reduces the cost and is beneficial to keeping the intact bioactivity of collagen.
Example 1 preparation of sterile collagen solution of adipose-derived stem cell exosomes
Mixing the adipose stem cell exosomes and sterile collagen liquid in a sterile environment, wherein each milliliter of the sterile collagen liquid contains 0.5mg of the exosomes, uniformly stirring, and storing at low temperature to obtain the sterile collagen liquid of the adipose stem cell exosomes.
Wherein the aseptic collagen liquid is fish skin collagen liquid; the adipose-derived stem cell exosome is extracted by the existing method.
Example 2 preparation of adipose-derived stem cell exosomes by sterile collagen liquid application
The method specifically comprises the following steps:
step 1, under the environment with aseptic process control, two rectangular non-woven fabrics with the same size and specification are taken, the aseptic collagen liquid of the adipose-derived stem cell exosome prepared in the embodiment 1 is coated between one non-woven fabric, and the coating thickness of the aseptic collagen liquid of the adipose-derived stem cell exosome is 2 mm;
step 2, coating a layer of viscose on the periphery of the non-woven fabric coated with the aseptic collagen liquid of the adipose-derived stem cell exosomes, wherein the thickness of the viscose is 0.5mm, and then bonding the two non-woven fabrics together to ensure that the aseptic collagen liquid of the adipose-derived stem cell exosomes is pressed between the two non-woven fabrics to obtain the aseptic collagen liquid application;
and 3, pasting the sterile collagen liquid obtained in the step 2 in an aluminum foil bag, sealing the aluminum foil bag, and storing the aluminum foil bag at a low temperature of 4 ℃.
Example 3 preparation of sterile collagen solution of adipose-derived stem cell exosomes
Mixing the adipose stem cell exosomes and sterile collagen liquid in a sterile environment, wherein each milliliter of the sterile collagen liquid contains 0.6mg of the exosomes, uniformly stirring, and storing at low temperature to obtain the sterile collagen liquid of the adipose stem cell exosomes.
Wherein the aseptic collagen liquid is fish skin collagen liquid; the adipose-derived stem cell exosome is extracted by the existing method.
Example 4 preparation of adipose Stem cell exosomes by sterile collagen liquid application
The method specifically comprises the following steps:
step 1, under the environment with aseptic process control, two rectangular non-woven fabrics with the same size and specification are taken, the aseptic collagen liquid of the adipose-derived stem cell exosome prepared in the embodiment 3 is coated between one non-woven fabric, and the coating thickness of the aseptic collagen liquid of the adipose-derived stem cell exosome is 2.5 mm;
step 2, coating a layer of viscose on the periphery of the non-woven fabric coated with the aseptic collagen liquid of the adipose-derived stem cell exosomes, wherein the thickness of the viscose is 1mm, and then bonding the two non-woven fabrics together to enable the aseptic collagen liquid of the adipose-derived stem cell exosomes to be pressed between the two non-woven fabrics to obtain the aseptic collagen liquid application;
and 3, pasting the sterile collagen liquid obtained in the step 2 in an aluminum foil bag, sealing the aluminum foil bag, and storing the aluminum foil bag at a low temperature of 4 ℃.
Example 5 preparation of sterile collagen solution of adipose-derived stem cell exosomes
Mixing the adipose stem cell exosomes with sterile collagen liquid in a sterile environment, wherein each milliliter of the sterile collagen liquid contains 0.7mg of the exosomes, uniformly stirring, and storing at low temperature to obtain the sterile collagen liquid of the adipose stem cell exosomes.
Wherein the aseptic collagen liquid is fish skin collagen liquid; the adipose-derived stem cell exosome is extracted by the existing method.
Example 6 preparation of adipose Stem cell exosomes by sterile collagen liquid application
The method specifically comprises the following steps:
step 1, under the environment with aseptic process control, two rectangular non-woven fabrics with the same size and specification are taken, the aseptic collagen liquid of the adipose-derived stem cell exosome prepared in the embodiment 5 is coated between one non-woven fabric, and the coating thickness of the aseptic collagen liquid of the adipose-derived stem cell exosome is 3 mm;
step 2, coating a layer of viscose on the periphery of the non-woven fabric coated with the aseptic collagen liquid of the adipose-derived stem cell exosomes, wherein the thickness of the viscose is 0.8mm, and then bonding the two non-woven fabrics together to ensure that the aseptic collagen liquid of the adipose-derived stem cell exosomes is pressed between the two non-woven fabrics to obtain the aseptic collagen liquid application;
and 3, pasting the sterile collagen liquid obtained in the step 2 in an aluminum foil bag, sealing the aluminum foil bag, and storing the aluminum foil bag at a low temperature of 4 ℃.
Example 7 preparation of sterile collagen solution of adipose-derived stem cell exosomes
Mixing the adipose stem cell exosomes and sterile collagen liquid in a sterile environment, wherein each milliliter of the sterile collagen liquid contains 0.8mg of the exosomes, uniformly stirring, and storing at low temperature to obtain the sterile collagen liquid of the adipose stem cell exosomes.
Wherein the aseptic collagen liquid is fish skin collagen liquid; the adipose-derived stem cell exosome is extracted by the existing method.
Example 8 preparation of adipose Stem cell exosomes by sterile collagen liquid application
The method specifically comprises the following steps:
step 1, under the environment with aseptic process control, two rectangular non-woven fabrics with the same size and specification are taken, the aseptic collagen liquid of the adipose stem cell exosome prepared in the embodiment 7 is coated between one non-woven fabric, and the coating thickness of the aseptic collagen liquid of the adipose stem cell exosome is 2.5 mm;
step 2, coating a layer of viscose on the periphery of the non-woven fabric coated with the aseptic collagen liquid of the adipose-derived stem cell exosomes, wherein the thickness of the viscose is 0.6mm, and then bonding the two non-woven fabrics together to ensure that the aseptic collagen liquid of the adipose-derived stem cell exosomes is pressed between the two non-woven fabrics to obtain the aseptic collagen liquid application;
and 3, pasting the sterile collagen liquid obtained in the step 2 in an aluminum foil bag, sealing the aluminum foil bag, and storing the aluminum foil bag at a low temperature of 4 ℃.
The aseptic collagen liquid of the adipose-derived stem cell exosome can be applied to skin injury repair and wound repair, such as burn wounds, scald wounds, laser beauty and the like.
Cytotoxicity assays
The results of the biological performance tests of the aseptic collagen liquid application of the adipose-derived stem cell exosomes prepared in the examples 2, 4, 6 and 8 show that the cytotoxicity score is less than 1, and the aseptic collagen liquid application has no sensitization reaction, no skin irritation reaction or extremely light micro irritation, no sub-chronic systemic toxicity and no genetic toxicity.
Detection of microbial performance
The results of microbiological examination of the aseptic collagen liquid application of the adipose stem cell exosomes prepared in examples 2, 4, 6 and 8 show sterility.
Detection of wound repair action
The results of the clinical experiments of the control, which were performed by applying the sterile collagen solution of the adipose stem cell exosomes prepared in the examples 2, 4, 6 and 8 to treat the superficial II-degree burn wounds randomly: the experimental sample size is 150 cases, the experimental apparatus is aseptic collagen liquid application of the adipose stem cell exosomes prepared in the embodiments 2, 4, 6 and 8, and the comparison apparatus is vaseline application dressing and aseptic collagen liquid application dressing, and the experimental result shows that compared with the vaseline application dressing and the aseptic collagen liquid application dressing, the aseptic collagen liquid application of the adipose stem cell exosomes prepared in the embodiments 2, 4, 6 and 8 can shorten the healing time of the II-degree burn wound faster, improve local symptom signs, promote the healing of the wound and improve the comprehensive healing rate of the wound, so that the pain of a patient in the wound repair process is relieved, and the clinical popularization and application are worthy.
While the foregoing description shows and describes several preferred embodiments of the invention, it is to be understood, as noted above, that the invention is not limited to the forms disclosed herein, but is not to be construed as excluding other embodiments and is capable of use in various other combinations, modifications, and environments and is capable of changes within the scope of the inventive concept as expressed herein, commensurate with the above teachings, or the skill or knowledge of the relevant art. And that modifications and variations may be effected by those skilled in the art without departing from the spirit and scope of the invention as defined by the appended claims.
Claims (8)
1. The aseptic collagen liquid of the adipose-derived stem cell exosome is characterized by consisting of the adipose-derived stem cell exosome and the aseptic collagen liquid, wherein each milliliter of the aseptic collagen liquid contains 0.5-0.8mg of exosome.
2. The sterile collagen fluid of an adipose stem cell exosome according to claim 1, wherein the sterile collagen fluid is fish skin collagen fluid.
3. The method for preparing the aseptic collagen liquid of the adipose-derived stem cell exosomes according to claim 1 or 2, characterized in that the adipose-derived stem cell exosomes and the aseptic collagen liquid are mixed under an aseptic environment, wherein each milliliter of the aseptic collagen liquid contains 0.5-0.8mg of the exosomes, and the aseptic collagen liquid of the adipose-derived stem cell exosomes is obtained by uniformly stirring and storing at a low temperature.
4. The method for preparing a sterile collagen liquid for an adipose-derived stem cell exosome according to claim 3, wherein the sterile collagen liquid is a fish skin collagen liquid.
5. The preparation method of the aseptic collagen liquid application of the adipose-derived stem cell exosome is characterized by comprising the following steps:
step 1, under the environment with aseptic process control, two rectangular non-woven fabrics with the same size and specification are taken, and the aseptic collagen liquid of the adipose-derived stem cell exosome prepared according to the claim 3 or 4 is coated between one of the non-woven fabrics;
step 2, coating a layer of viscose on the periphery of the non-woven fabric coated with the aseptic collagen liquid of the adipose-derived stem cell exosomes, and then bonding the two non-woven fabrics together to enable the aseptic collagen liquid of the adipose-derived stem cell exosomes to be pressed between the two non-woven fabrics, so that the aseptic collagen liquid is obtained and applied;
and 3, sticking the sterile collagen liquid obtained in the step 2 in an aluminum foil bag, sealing the aluminum foil bag, and storing the aluminum foil bag at a low temperature.
6. The method for preparing the aseptic collagen liquid application of the adipose stem cell exosomes according to claim 5, wherein the aseptic collagen liquid application thickness of the adipose stem cell exosomes in the step 1 is 2-3 mm.
7. The method for preparing an adipose stem cell exosome-containing sterile collagen liquid application according to claim 5 or 6, wherein the thickness of the mucilage glue in the step 2 is 0.5-1 mm.
8. Use of a sterile collagen liquid of an adipose stem cell exosome prepared according to any one of claims 5 to 7 in the preparation of a skin injury repair product, a wound repair product, a medical cosmetic product or a cosmetic product.
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