CN112891632B - Preparation method of human umbilical cord intima-fibrin composite skin scaffold with immunogen removed - Google Patents
Preparation method of human umbilical cord intima-fibrin composite skin scaffold with immunogen removed Download PDFInfo
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- 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
- A61L27/14—Macromolecular materials
- A61L27/22—Polypeptides or derivatives thereof, e.g. degradation products
- A61L27/225—Fibrin; Fibrinogen
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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
- A61L27/14—Macromolecular materials
- A61L27/22—Polypeptides or derivatives thereof, e.g. degradation products
- A61L27/227—Other specific proteins or polypeptides not covered by A61L27/222, A61L27/225 or A61L27/24
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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
- A61L27/50—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
- A61L2300/20—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices containing or releasing organic materials
- A61L2300/252—Polypeptides, proteins, e.g. glycoproteins, lipoproteins, cytokines
- A61L2300/254—Enzymes, proenzymes
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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
- A61L2300/418—Agents promoting blood coagulation, blood-clotting agents, embolising 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
- A61L2430/00—Materials or treatment for tissue regeneration
- A61L2430/34—Materials or treatment for tissue regeneration for soft tissue reconstruction
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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
- A61L2430/00—Materials or treatment for tissue regeneration
- A61L2430/40—Preparation and treatment of biological tissue for implantation, e.g. decellularisation, cross-linking
Abstract
The invention discloses a preparation method of a deimmunogen human umbilical cord intima-fibrin composite skin stent, which belongs to the technical field of artificial skin and comprises the following steps: taking placenta without virus and bacterial infection, cutting off umbilical cord, performing non-invasive expansion method with sterile normal saline, air, nitrogen, etc. under sterile condition, performing blunt dissection from umbilical cord basal layer to obtain umbilical cord inner membrane, and removing impurities on the surface of umbilical cord inner membrane; cleaning the stripped umbilical cord intima with normal saline, placing the umbilical cord intima into a self-prepared culture solution containing antibiotics for soaking and disinfection, filtering and subpackaging the soaking solution, and storing at-80 ℃ for later use; the inner membrane of the umbilical cord is dehydrated by using organic solution-physiological saline with different concentrations. The skin bracket attached to the wound has activity, so that the skin bracket can be integrated with the skin of a human body within a period of time after being attached to the affected part of the wound, and the wound can be permanently repaired.
Description
Technical Field
The invention relates to the technical field of artificial skin, in particular to a human umbilical cord intima-fibrin composite skin scaffold for removing immunogen.
Background
The umbilical cord intima of the matrix layer is a thin-film matrix formed by connecting single-layer epithelial cells. Along with the growth and development of the embryo, the umbilical cord intima and the umbilical cord matrix layer grow in an attached manner and are rich in toughness. The umbilical cord inner membrane is the innermost layer of the umbilical cord, is smooth, has no blood vessels, no nerves and lymph, has certain elasticity and is about 0.1-0.2mm thick, contains collagen, fibronectin, laminin and other components, and the components enable the umbilical cord inner membrane to serve as a 'transplanted basement membrane' to play a new and proper matrix role in health to promote epithelization. Has the characteristics of promoting the adhesion growth and proliferation of epithelium, relieving inflammation, inhibiting the formation of new vessels, reducing scar hyperplasia, resisting adhesion and the like, is applied to severe acute or old skin surface burns and skin defect wounds, provides a colloid bracket containing a substrate and a matrix component, and ensures that the proliferated epithelial cells can be expanded and migrated on the colloid bracket; moreover, the umbilical cord intima is helpful for proliferation of stem cells and provides a microenvironment suitable for growth of the stem cells, the umbilical cord intima is used as a novel biological material, a miraculous effect is achieved, (the experimental results of large and small animals prove that scars are small, and fur grows), and the umbilical cord intima is wide in source, low in price, simple and convenient to obtain and store, simple in operation and almost free of immunological rejection reaction, and very suitable for being developed in primary hospitals.
The skin is planted to human affected part at present, the majority adopts autologous skin to transplant, but when the wound area is great, the removal of autologous skin is limited, thereby can not satisfy medical demand, and because the transplantation of autologous skin need can be realized through the operation, this will increase the patient's misery during the treatment, be unfavorable for the healing of patient's wound, in addition although appear on the market now and be used for the product of clinical skin, but because it does not have the ductility, lead to synthetic skin support to cut the fixed back of size, the subsides that only can be applicable to the affected part of certain size apply, this will influence the application scope of compound back skin support.
Disclosure of Invention
1. Technical problem to be solved
The invention aims to provide a preparation method of a deimmunized human umbilical cord intima-fibrin composite skin scaffold, which aims to solve the problems in the background technology.
2. Technical scheme
In order to solve the problems, the invention adopts the following technical scheme:
the preparation method of the deimmunized human umbilical cord intima-fibrin composite skin scaffold comprises the following steps:
1) Taking placenta without virus and bacterial infection, cutting off umbilical cord, performing non-invasive expansion method with sterile normal saline, air, nitrogen, etc. under sterile condition, performing blunt dissection from umbilical cord basal layer to obtain umbilical cord inner membrane, and removing impurities on the surface of umbilical cord inner membrane;
2) Cleaning the stripped umbilical cord inner membrane with physiological saline containing 0.1-1mol/L NaOH for 3 times, soaking in self-prepared culture solution of antibiotics for disinfection, filtering the soaking solution, subpackaging, and storing at-80 deg.C for use;
3) Transferring the umbilical cord intima into 0.5-3mol/L NaOH solution to be soaked for 24-72h for sterilization, and then dehydrating by using organic solution-normal saline with different concentrations;
4) Placing the dehydrated umbilical cord inner membrane in a sterile ultra-clean workbench, flatly paving the umbilical cord inner membrane at the bottom of a sterile cell culture dish, and standing;
5) Uniformly smearing a solution containing 0.1-1000U/ml human thrombin and 2-20% human serum albumin on the surface of the umbilical cord intima matrix after immunogen removing treatment;
6) Uniformly coating a solution containing 2.0-5.0mg/ml of equal volume of fibrinogen on the surface of the treated umbilical cord intima matrix;
7) Adding a culture dish cover, transferring into a constant-temperature drying oven at 20-45 ℃, and standing for 0.5-24h to obtain a dry umbilical cord intima-fibrin composite skin stent;
8) The umbilical cord intima-fibrin composite skin scaffold can be cut into umbilical cord intima-fibrin composite skin scaffolds with different specifications as required in a sterile room;
9) Carrying out sterile vacuum packaging, drying in the shade, and storing at room temperature for later use;
10 Before use, the umbilical cord intima-fibrin composite skin stent is taken out in advance and soaked in the umbilical cord intima-fibrin composite skin stent renaturation culture solution which is separately stored, and the umbilical cord intima-fibrin composite skin stent renaturation culture solution can be used for patients with skin defect after 0.5-24 h.
Preferably, the umbilical cord in step 1) is derived from a human body.
Preferably, the antibiotics in the step 2) adopt liquid broad-spectrum antibiotics, and the soaking disinfection time of the umbilical cord intima is 0.5-72h.
Preferably, the self-prepared culture solution in step 2) is derived from a modified commercial umbilical cord intimal cell culture medium and specifically LF-BIO, MSCM supplemented with serum replacement.
Preferably, the step 2) adopts a 0.25um filter membrane for filtration.
Preferably, the step 3) adopts methanol-physiological saline with different concentrations to perform gradient dehydration on the umbilical cord intima.
3. Advantageous effects
1. The invention adopts two smearing processes, thrombin, human serum albumin and fibrin solution with controllable dosage are respectively and evenly smeared on the surface of the umbilical cord intima-fibrin composite skin bracket, and the prepared umbilical cord intima-fibrin composite skin bracket can protect the attached wound through the human serum albumin on the skin bracket when the skin bracket is attached to the wound, thereby reducing the probability of the wound having ulceration phenomenon, simultaneously providing nutrient components for the damaged tissue and promoting the healing of the wound.
2. The umbilical cord intima-fibrin composite patch is independently stored in the culture solution prepared in the step 2 before use, so that the umbilical cord intima-fibrin composite patch can be effectively and quickly softened, the adhesion of a composite scaffold is convenient, meanwhile, various original cytokines can be attached to the renatured umbilical cord intima-fibrin composite patch (artificial skin), the umbilical cord intima-fibrin composite patch can be directly applied to an affected part to help the wound to quickly heal, and skin flaps are fused into pieces, so that scars are small and flat, the surface of the skin without protrusions is fused with normal skin, and the regeneration of hair follicles can be promoted;
3. the skin stent attached to the wound has activity, so that the skin stent can be integrated with the skin of a human body within a period of time after being attached to the affected part of the wound, the wound can be permanently repaired, the condition that the skin of the affected part can be better restored only by carrying out skin grafting on the affected part for many times at present is avoided, and the pain of a patient caused by repeated skin grafting can be relieved by the composite skin stent.
4. The invention has wide material source, is the post-partum waste, has lower cost, is convenient for the manufacture of artificial skin and effectively reduces the contradiction between supply and demand of the skin; in addition, the composite skin stent has the advantages of convenient material acquisition, simple manufacturing process, convenient operation of medical personnel, convenient storage, convenient transportation, convenient commercial preparation and clinical application.
5. The invention can preserve the integrity of the umbilical cord intima matrix to a large extent by treating the umbilical cord intima, so that the prepared umbilical cord intima has extensibility and plasticity, and medical workers can apply the composite skin stent to an affected part in an extension mode according to the wound proportion, thereby improving the applicability of the composite skin stent.
6. The invention leads the compounded skin stent to have good compatibility and no rejection reaction by removing the allocollagen and the related immunogen from the umbilical cord intima so as to ensure the use safety of the compounded skin stent.
7. The invention can further enlarge the extension area of the umbilical cord intima by combining with mechanical expansion, and further improve the flexibility and elasticity so as to better realize wound fusion.
Drawings
FIG. 1 is a schematic representation of the treatment of porcine skin in accordance with the present invention; in the figure, A is: the scald area of the male small-ear pig is 11 x 12cm after deep III-degree scald; b: the wound is disinfected after 1 day of scald; c: large-area infection and fester appear on the wound after 5 days of scald; d, scalding for 11 days, and dying the auricle pig due to severe infection; e, scalding the male small-ear pig at the deep III degree with the scald area of 10 x 13.2cm; f: starting the operation 3 days after the scald, and implanting a skin bracket into the wound with the wound depth of 1.5cm to the myofascial layer; g, 20 days after the scaffold is implanted, the wound is free of infection, dry, ruddy, compatible with tissues and free of rejection reaction, the wound area is 1 x 2cm, and hair grows around the wound; h is: after the scaffold is implanted for 65 days, the wound is completely healed, and the hair is dense and glossy;
fig. 2 is a diagram showing comparison before and after stretching and wound plasticity of an umbilical cord intima in a ductility test.
Detailed Description
The present invention will be described in further detail with reference to the accompanying drawings and examples.
Examples
The preparation method of the deimmunized human umbilical cord intima-fibrin composite skin scaffold comprises the following steps:
1) Selecting an in vitro placenta, cutting off an umbilical cord, detecting by serum to eliminate virus and bacterial infections, such as HIV, syphilis, HBV, HCV and the like, carrying out blunt stripping on the umbilical cord substrate under an aseptic condition to obtain an umbilical cord intima, and removing impurities on the surface of the umbilical cord intima, wherein the umbilical cord is from a human body;
2) Washing the stripped umbilical cord intima for 3 times by using normal saline containing 0.1-1mol/L NaOH, then placing the umbilical cord intima into self-prepared culture solution of antibiotics for soaking and disinfection, then filtering and subpackaging the soaking solution, and placing the soaked solution at-80 ℃ for storage and standby use, wherein a 0.25um filter membrane is adopted for filtering, the antibiotics adopt liquid broad-spectrum antibiotics, the soaking and disinfection time of the umbilical cord intima is 0.5-24h, the self-prepared culture solution is from an improved commercialized umbilical cord intima cell culture medium and specifically is LF-BIO, MSCM, and the culture medium is a basic cell culture medium added with the following components: 2-5% by volume of a clinical-grade non-serum substitute.
3) Transferring the umbilical cord inner membrane into 0.5-3mol/L NaOH solution, soaking for 24-72h for sterilization, and then dehydrating with organic solution-physiological saline with different concentrations, wherein the organic solution-physiological saline is specifically methanol-physiological saline;
4) Placing the dehydrated umbilical cord intima in a sterile ultra-clean workbench, paving at the bottom of a sterile cell culture dish, and standing;
5) Uniformly coating a solution containing 0.1-1000U/ml human thrombin and 2-20% human serum protein on the surface of the umbilical cord intima matrix after immunogen removing treatment;
6) Uniformly coating a solution containing 2.0-5.0mg/ml of equal volume of fibrinogen on the surface of the treated umbilical cord intima matrix;
7) Adding a culture dish cover, transferring into a constant-temperature drying oven at 20-45 ℃, and standing for 0.5-24h to obtain a dried umbilical cord intima-fibrin composite skin scaffold;
8) In the superclean workbench, the umbilical cord intima-fibrin composite skin scaffolds with different specifications can be cut according to requirements;
9) Carrying out sterile vacuum packaging, drying in the shade, and storing at room temperature for later use;
10 Before use, the umbilical cord intima-fibrin composite skin stent is taken out in advance and soaked in the umbilical cord intima-fibrin composite skin stent renaturation culture solution which is separately stored, and the umbilical cord intima-fibrin composite skin stent renaturation culture solution can be used for patients with skin defect after 0.5-24 h.
The following test for correlation performance of the umbilical cord intima-fibrin composite skin scaffold, as shown in figure 2:
1. ductility: original flat laying 3.5cm, 8.5cm after pulling open and extending about 0.1-0.2mm thick;
2. plasticity: can be applied to the affected part according to the shape of the wound surface, is convenient to use and saves the cost.
Application example 1
By adopting the preparation method of example 1, the umbilical cord intima-fibrin composite skin scaffold is prepared, then a skin burn model is selected, the animal number is 152700, and male Yunnan small ear pigs with the age of 114 days and the weight of 11.9kg are planted in the burn position, and the umbilical cord intima-fibrin composite skin scaffold is finally cured.
Specifically, as shown in fig. 1, fig. 1 shows the treatment process of the pigskin according to the invention; in the figure, A is: scalding male small-ear pigs at deep III degree with the scald area of 11 x 12cm; b: 1 day of scald treatment, and sterilizing the wound; c: after 5 days of scald, large-area infection and fester appear on the wound; d, scalding for 11 days, and dying the auricle pig due to severe infection; e, scalding the male small-ear pig at the deep III degree with the scald area of 10 x 13.2cm; f: starting the operation 3 days after the scald, wherein the wound depth is 1.5cm to the myofascial layer, and implanting a skin stent; g, 20 days after the stent is implanted, the wound is free of infection, dry, ruddy, compatible with tissues and free of rejection reaction, the area of the wound is 1 x 2cm, and hair grows around the wound; h is: after the stent is implanted for 65 days, the wound is completely healed, and the hair is dense and glossy.
It should be understood by those skilled in the art that the above embodiments are only for illustrating the present invention and are not to be used as a limitation of the present invention, and that changes and modifications to the above embodiments are within the scope of the claims of the present invention as long as they are within the spirit and scope of the present invention.
Claims (3)
1. The preparation method of the deimmunized human umbilical cord intima-fibrin composite skin scaffold is characterized by comprising the following steps of:
1) Taking placenta without virus and bacterial infection, cutting off umbilical cord which is from human body, carrying out non-invasive expansion method by using sterile normal saline, air and nitrogen under sterile conditions to obtain umbilical cord intima from umbilical cord basal layer, and removing impurities on umbilical cord intima surface;
2) Washing the stripped umbilical cord intima with physiological saline containing 0.1-1mol/L NaOH for 3 times, soaking and sterilizing in a self-prepared culture solution of antibiotics, filtering and subpackaging the soaked solution, and storing at-80 ℃ for later use, wherein the self-prepared culture solution is prepared from an improved commercialized umbilical cord intima cell culture medium, specifically LF-BIO, MSCM Cat No: LF0101A with 2-5% serum replacement;
3) Transferring the umbilical cord intima into 0.5-3mol/L NaOH solution to be soaked for 24-72h for sterilization treatment, and then performing gradient dehydration on the umbilical cord intima by adopting methanol-physiological saline with different concentrations;
4) Placing the dehydrated umbilical cord inner membrane in a sterile ultra-clean workbench, flatly paving the umbilical cord inner membrane at the bottom of a sterile cell culture dish, and standing;
5) Uniformly smearing a solution containing 0.1-1000U/ml of human thrombin and 2-20% of human serum albumin on the surface of the umbilical cord intima matrix after immunogen removing treatment;
6) Uniformly coating a solution containing 2.0-5.0mg/ml of equal volume of fibrinogen on the surface of the treated umbilical cord intima matrix;
7) Adding a culture dish cover, transferring into a constant-temperature drying oven at 20-45 ℃, and standing for 0.5-24h to obtain a dry umbilical cord intima-fibrin composite skin scaffold;
8) The umbilical cord intima-fibrin composite skin scaffold with different specifications can be cut into umbilical cord intima-fibrin composite skin scaffolds with different specifications as required in a sterile room;
9) And carrying out sterile vacuum packaging, drying in the shade, and storing at room temperature for later use.
2. The method for preparing the deimmunized human umbilical cord intima-fibrin composite skin scaffold according to claim 1, wherein the antibiotics in the step 2) are liquid broad spectrum antibiotics, and the soaking disinfection time of the umbilical cord intima is 0.5-72h.
3. The method for preparing the deimmunized human umbilical cord intima-fibrin composite skin scaffold according to claim 1, wherein the step 2) is performed by filtration with a sterile filter membrane.
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| US20080118551A1 (en) * | 2006-11-20 | 2008-05-22 | Yasmin Wadia | Denatured human albumin biomaterial as dressing for burns and wounds of skin |
| IL207586A0 (en) * | 2010-08-12 | 2010-12-30 | Omrix Biopharmaceuticals Ltd | A fibrin based therapeutic preparation and use thereof |
| CN110404119B (en) * | 2019-08-29 | 2023-04-07 | 上海越增生物科技有限公司 | Preparation method of amnion tissue engineering immunogen-removing skin scaffold |
| CN112263713A (en) * | 2020-09-21 | 2021-01-26 | 苏州大学附属第一医院 | Fibrin hydrogel scaffold loaded with human umbilical cord mesenchymal stem cells and application thereof |
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