CN113509595A - Dermal substitute and preparation method thereof - Google Patents
Dermal substitute and preparation method thereof Download PDFInfo
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- CN113509595A CN113509595A CN202110938433.1A CN202110938433A CN113509595A CN 113509595 A CN113509595 A CN 113509595A CN 202110938433 A CN202110938433 A CN 202110938433A CN 113509595 A CN113509595 A CN 113509595A
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- A—HUMAN NECESSITIES
- 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
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
- A61L27/3604—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix characterised by the human or animal origin of the biological material, e.g. hair, fascia, fish scales, silk, shellac, pericardium, pleura, renal tissue, amniotic membrane, parenchymal tissue, fetal tissue, muscle tissue, fat tissue, enamel
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- A—HUMAN NECESSITIES
- 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
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/02—Inorganic materials
- A61L27/04—Metals or alloys
- A61L27/047—Other specific metals or alloys not covered by A61L27/042 - A61L27/045 or A61L27/06
-
- A—HUMAN NECESSITIES
- 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
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/14—Macromolecular materials
- A61L27/22—Polypeptides or derivatives thereof, e.g. degradation products
- A61L27/24—Collagen
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- A—HUMAN NECESSITIES
- 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
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
- A61L27/3683—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix subjected to a specific treatment prior to implantation, e.g. decellularising, demineralising, grinding, cellular disruption/non-collagenous protein removal, anti-calcification, crosslinking, supercritical fluid extraction, enzyme treatment
- A61L27/3687—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix subjected to a specific treatment prior to implantation, e.g. decellularising, demineralising, grinding, cellular disruption/non-collagenous protein removal, anti-calcification, crosslinking, supercritical fluid extraction, enzyme treatment characterised by the use of chemical agents in the treatment, e.g. specific enzymes, detergents, capping agents, crosslinkers, anticalcification agents
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- A—HUMAN NECESSITIES
- 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
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
- A61L27/3683—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix subjected to a specific treatment prior to implantation, e.g. decellularising, demineralising, grinding, cellular disruption/non-collagenous protein removal, anti-calcification, crosslinking, supercritical fluid extraction, enzyme treatment
- A61L27/3691—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix subjected to a specific treatment prior to implantation, e.g. decellularising, demineralising, grinding, cellular disruption/non-collagenous protein removal, anti-calcification, crosslinking, supercritical fluid extraction, enzyme treatment characterised by physical conditions of the treatment, e.g. applying a compressive force to the composition, pressure cycles, ultrasonic/sonication or microwave treatment, lyophilisation
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- A—HUMAN NECESSITIES
- 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
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
- A61L27/54—Biologically active materials, e.g. therapeutic substances
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- A—HUMAN NECESSITIES
- 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
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
- A61L27/60—Materials for use in artificial skin
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- A—HUMAN NECESSITIES
- 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
- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/10—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices containing or releasing inorganic materials
- A61L2300/102—Metals or metal compounds, e.g. salts such as bicarbonates, carbonates, oxides, zeolites, silicates
- A61L2300/104—Silver, e.g. silver sulfadiazine
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- A—HUMAN NECESSITIES
- 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
- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/40—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a specific therapeutic activity or mode of action
- A61L2300/404—Biocides, antimicrobial agents, antiseptic agents
Abstract
The invention relates to a dermis substitute which comprises the following components in parts by weight: 80-99.95 parts of acellular matrix fiber; 0.05-20 parts of collagen; 0-3 parts of silver; 60-80 parts of acellular biological membrane. The invention also relates to a method for preparing the dermal substitute. The outer layer of the dermal substitute is an acellular biological membrane, so that the wound surface can be protected, the excessive evaporation of water on the wound surface can be prevented, and compared with a high-molecular membrane such as a silica gel membrane, the dermal substitute has the air permeability closer to the original function of the skin; the inner layer is acellular matrix fiber which can replace the defective dermis and promote the growth of granulation so as to form a substrate for secondary implantation into epidermis; the dispersed silver ions can inhibit the colonization of microorganisms and increase the recovery effect of the wound surface.
Description
Technical Field
The invention relates to the technical field of biological materials, in particular to a dermis substitute and a preparation method thereof.
Background
At present, full-thickness skin defects caused by burns, trauma, excision, skin flap removal and the like can be applied to wound healing by using a dermal substitute.
Currently, a polymer film (such as a silicon film) is usually adopted as an outer layer film for the genuine leather substitute, so that the ventilation capacity of the genuine leather substitute cannot be close to the original function of the skin; furthermore, dermal substitutes are used at risk of colonization by such microorganisms, which may affect the recovery of the wound and may even cause infections.
Thus, there is a need for a dermal substitute and method of making the same.
Disclosure of Invention
The invention aims to provide a dermis substitute and a preparation method thereof aiming at the defects in the prior art.
In order to achieve the purpose, the invention adopts the technical scheme that:
the first aspect of the invention provides a dermis substitute, which comprises the following components in parts by weight:
a second aspect of the present invention provides a method for preparing a dermal substitute, as described above, comprising the steps of:
s1, taking the wet acellular matrix, shearing the matrix into fragments, and then mashing the fragments at a high speed in a short time;
s2, taking the acellular matrix, glacial acetic acid and purified water, mixing and cooling;
s3, homogenizing at a certain frequency, filtering coarse particles after homogenizing the collected feed liquid for a plurality of times, adjusting the pH value of the filtered slurry by adopting a NaOH solution, and centrifuging to obtain the acellular matrix fiber;
s4, uniformly mixing the acellular matrix fibers and the collagen, preparing a water system dispersion, adjusting the pH value to 4-8, uniformly mixing and dispersing, covering the acellular biological membrane, performing vacuum freeze drying, cutting, packaging and sterilizing to obtain the dermis substitute.
Preferably, in step S1, the cut pieces are smaller than 1cm2The high speed mashing time is not more than 1 min.
Preferably, in step S2, 0.5-3.0% of the acellular matrix, 0.01-0.1% of glacial acetic acid, and the balance of purified water are mixed and cooled to 2-8 ℃.
Preferably, in step S3, the frequency of the homogenate is in the range of 30Hz to 35 Hz.
Preferably, in step S3, the total number of homogenations is 6.
Preferably, in step S3, the concentration of the NaOH solution used is 8% -10%.
Preferably, in step S3, the pH of the slurry after adjustment is 7.5-8.0.
Preferably, in step S4, after adjusting the pH to 4 to 8, the silver salt solution or salt dispersion is added according to the solid content, and then mixed and dispersed uniformly.
Preferably, in step S4, the silver salt includes: at least one of silver sulfate, silver chloride, silver sulfadiazine, silver oxide or nano silver.
By adopting the technical scheme, compared with the prior art, the invention has the following technical effects:
the outer layer of the dermal substitute is an acellular biological membrane, so that the wound surface can be protected, the excessive evaporation of water on the wound surface can be prevented, and compared with a high-molecular membrane such as a silica gel membrane, the dermal substitute has the air permeability closer to the original function of the skin; the inner layer is acellular matrix fiber which can replace the defective dermis and promote the growth of granulation so as to form a substrate for secondary implantation into epidermis; the dispersed silver ions can inhibit the colonization of microorganisms and increase the recovery effect of the wound surface.
Drawings
Fig. 1 is a schematic structural view of the dermal substitute prepared in example 1 of the present invention.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious 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.
It should be noted that the embodiments and features of the embodiments may be combined with each other without conflict.
The invention is further described with reference to the following drawings and specific examples, which are not intended to be limiting.
Example 1
As shown in fig. 1, the present example provides a dermal substitute prepared by the steps of:
s1, taking the wet acellular matrix, and shearing the matrix into pieces smaller than 1cm2Crushing at high speed in a short time (not more than 1 min);
s2, taking 0.5% (wet weight) of the acellular matrix, 0.01% of glacial acetic acid and the balance of purified water according to the weight percentage, mixing and cooling to 2 ℃;
s3, homogenizing at the frequency of 30Hz, homogenizing the collected feed liquid for 5 times, filtering out coarse particles, adjusting the pH value of the filtered slurry to 7.5 by adopting a NaOH solution with the concentration of 8%, and centrifuging to obtain the acellular matrix fiber;
s4, taking 80 parts of the acellular matrix fiber and 20 parts of collagen, mixing uniformly to prepare a water system dispersion, adjusting the pH value to 4, mixing and dispersing uniformly, covering 60 parts of acellular biological membranes, vacuum freeze-drying, cutting, packaging and sterilizing to obtain the dermis substitute.
Example 2
This example provides another dermal substitute prepared by the steps comprising:
s1, taking the wet acellular matrix, and shearing the matrix into pieces smaller than1cm2Crushing at high speed in a short time (not more than 1 min);
s2, taking 3.0% (wet weight) of the acellular matrix, 0.1% of glacial acetic acid and the balance of purified water according to the weight percentage, mixing and cooling to 8 ℃;
s3, homogenizing at the frequency of 35Hz, homogenizing the collected feed liquid for 5 times, filtering out coarse particles, adjusting the pH value of the filtered slurry to 8.0 by adopting a NaOH solution with the concentration of 10%, and centrifuging to obtain the acellular matrix fiber;
and S4, uniformly mixing 99 parts of the acellular matrix fiber and 1 part of collagen, preparing a water system dispersion, adjusting the pH value to 8, adding 3 parts of nano silver dispersion according to the solid content, uniformly mixing and dispersing, covering 80 parts of acellular biological membranes, performing vacuum freeze drying, cutting, packaging and sterilizing to obtain the dermis substitute.
Example 3
Fixing the test animal on a rat fixer, and shaving off the peril hair in the operation area; sterilizing the surgical area with iodophor; 3% sodium pentobarbital is injected to anaesthetize the test animals; cutting epidermis to reach dermis layer, making damaged area of epidermis with size of about 1.5cm × 1.5cm, and cutting epidermis in the area; leaving a photograph of the wound for each animal; the dermal substitute prepared in example 1 was applied to the backs of the animals in the experimental group, and the time for hemostasis was observed and recorded; a control sample (plain medical gauze, i.e., sterile ethylene oxide sterilized gauze, such as suzuki standard 20162140204) was applied to the back of the control animal, and the time to hemostasis was observed and recorded; then, the experimental group and the control group cover sterile gauze on the dermis substitute; then fixing the medical adhesive tape by using elastic medical adhesive tape; wound area was observed daily: firstly, if the appearance of the dermis substitute cannot be observed visually, a new dermis substitute needs to be attached continuously until the wound is healed; observing whether the wound is whitish, indicating that an infection condition exists if the wound is whitish, cleaning the wound, replacing a dermis substitute and recording; if the appearance of the dermal substitute cannot be observed visually, new dermal substitute needs to be continuously fitted until the wound is healed; after healing after injury, recording healing time, observing scar conditions (taking pictures), finally killing animals, taking the skin of a test area for flattening and fixing, repairing, taking conventional paraffin for embedding the central part of the wound, slicing, carrying out conventional H-E staining, and observing healing conditions, inflammatory cell infiltration and blood vessels under a microscope.
As a result: after the dermal substitute prepared in example 1 was applied to the created wound, the bleeding of the wound was stopped immediately, and the test material was infiltrated with a small amount of blood and interstitial fluid, which rapidly forms a scab with the test material; the dermal substitute prepared in example 1 and the control product applied to the wound did not show infection symptoms such as bleeding or pus exudation during the whole experiment; after scabbing and falling off, white and soft new skin is exposed, and new quilt hair gradually grows.
Since only the epidermis layer of the experimental mouse is cut off and the dermis layer of the experimental mouse is not damaged in the embodiment, new hair gradually grows after the epidermis is healed; in this example, the wound is judged to be completely healed by the fact that the wound site begins to grow again. As shown in table 1, the wound healing time of the experimental group and the control group was about 20 days without significant difference, but 3 rats still had no significant hair growth at 27 days in the control group.
TABLE 1
| Observation items | Experimental group | Control group | P value |
| Average healing time (day) | 20.7±5.68 | 20.6±5.54 | 0.97 |
TABLE 2
As shown in Table 2, the wound was observed under a microscope to be completely healed without infiltration of red blood cells, inflammatory cells were almost zero, no fibrosis trace was observed, angiogenesis was observed between epidermal cells and stromal cells in a part of the sections, and clear hair follicles and root hair were observed in individual sections. The flat multiple layer epidermal cells can be clearly seen on the whole to cover on the round basal cells to form a complete skin structure.
In conclusion, the dermal substitute of the present invention can rapidly absorb the exuded blood and interstitial fluid and rapidly form a scab together with the exuded blood and interstitial fluid, and the scab skin can protect the wound from further infection, thereby eliminating the influence of infection and other factors on the wound healing time. In addition, the healing time of the wound covered by the invention and the wound covered by the control product has no obvious difference, but the experimental group is slightly better than the control group in the aspect of healing completion evaluation. The wound section is observed under the mirror, the wound covered by the invention has no obvious difference from the wound covered by the reference substance, and all the wounds form a complete skin structure with functionality.
While the invention has been described with reference to a preferred embodiment, it will be understood by those skilled in the art that various changes in form and detail may be made therein without departing from the spirit and scope of the invention.
Claims (10)
1. A dermal substitute, comprising, in parts by weight:
2. a method of preparing a dermal substitute according to claim 1, comprising the steps of:
s1, taking the wet acellular matrix, shearing the matrix into fragments, and then mashing the fragments at a high speed in a short time;
s2, taking the acellular matrix, glacial acetic acid and purified water, mixing and cooling;
s3, homogenizing at a certain frequency, filtering coarse particles after homogenizing the collected feed liquid for a plurality of times, adjusting the pH value of the filtered slurry by adopting a NaOH solution, and centrifuging to obtain the acellular matrix fiber;
s4, uniformly mixing the acellular matrix fibers and the collagen, preparing a water system dispersion, adjusting the pH value to 4-8, uniformly mixing and dispersing, covering the acellular biological membrane, performing vacuum freeze drying, cutting, packaging and sterilizing to obtain the dermis substitute.
3. The method of claim 2, wherein the cut pieces are less than 1cm in step S12The high speed mashing time is not more than 1 min.
4. The method according to claim 2, wherein in step S2, 0.5-3.0% of the acellular matrix, 0.01-0.1% of glacial acetic acid and the balance of purified water are mixed and cooled to 2-8 ℃.
5. The method according to claim 2, wherein the frequency of the homogenate in the step S3 is 30Hz to 35 Hz.
6. The method of claim 2, wherein the total number of homogenations in step S3 is 6.
7. The method of claim 2, wherein the NaOH solution is used at a concentration of 8% -10% in step S3.
8. The method according to claim 2, wherein the pH of the slurry after the adjustment in step S3 is 7.5 to 8.0.
9. The method according to claim 2, wherein in step S4, the silver salt solution or salt dispersion is added according to the solid content after adjusting the pH to 4 to 8, and then mixed and dispersed uniformly.
10. The production method according to claim 9, wherein in step S4, the silver salt includes: at least one of silver sulfate, silver chloride, silver sulfadiazine, silver oxide or nano silver.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110938433.1A CN113509595A (en) | 2021-08-16 | 2021-08-16 | Dermal substitute and preparation method thereof |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110938433.1A CN113509595A (en) | 2021-08-16 | 2021-08-16 | Dermal substitute and preparation method thereof |
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| CN113509595A true CN113509595A (en) | 2021-10-19 |
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| CN202110938433.1A Pending CN113509595A (en) | 2021-08-16 | 2021-08-16 | Dermal substitute and preparation method thereof |
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101361990A (en) * | 2008-09-03 | 2009-02-11 | 陕西瑞盛生物科技有限公司 | Double layer artificial skin and preparation method thereof |
| CN105999410A (en) * | 2016-05-05 | 2016-10-12 | 广州昕生医学材料有限公司 | Acellular tissue matrix composite and preparation method thereof |
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2021
- 2021-08-16 CN CN202110938433.1A patent/CN113509595A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101361990A (en) * | 2008-09-03 | 2009-02-11 | 陕西瑞盛生物科技有限公司 | Double layer artificial skin and preparation method thereof |
| CN105999410A (en) * | 2016-05-05 | 2016-10-12 | 广州昕生医学材料有限公司 | Acellular tissue matrix composite and preparation method thereof |
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Application publication date: 20211019 |