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A construction method and equivalent technology, applied in medical science, prosthesis, etc., can solve the problem of high shrinkage rate, and achieve the effect of promoting healing, increasing operability, and good compatibility
Active Publication Date: 2013-12-11
INST OF ANIMAL SCI OF CHINESE ACAD OF AGRI SCI
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[0005] The purpose of the present invention is to provide a method for constructing a dermis equivalent with human amniotic membrane and collagen as scaffolds for the existing defect of high shrinkage of the dermis equivalent made of collagen gel
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Embodiment 1
[0035] A construction method of a corium equivalent of the present invention, the detailed steps of the construction method are as follows:
[0036] a. Sterile processing of human amniotic membrane:
[0037]Pretreatment: The placenta was taken from the cesarean section of the puerpera, and the HIV, hepatitis A, hepatitis B, and syphilis serological reactions of the placenta were tested negative, and the tested placenta was placed in physiological saline with three antibodies for disinfection (adding three The normal saline used for the normal saline is 0.9% NaCl solution, and the three antibodies added are penicillin, streptomycin and gentamicin. After adding the normal saline, penicillin, streptomycin and gentamicin The concentrations of mycin are 0.1 μl / mL, 0.1 μl / mL and 0.1 μl / mL respectively), and the soaking and disinfection time is 3 to 4 hours;
[0038] Removal of the spongy layer: In a sterile room, bluntly peel off the human amniotic membrane attached to the pretreat...
Embodiment 2
[0046] A construction method of a corium equivalent of the present invention, the detailed steps of the construction method are as follows:
[0047] a. Sterile processing of human amniotic membrane:
[0048] Pretreatment: The placenta was taken from the cesarean section of the puerpera, and the HIV, hepatitis A, hepatitis B, and syphilis serological reactions of the placenta were tested negative, and the tested placenta was placed in physiological saline with three antibodies for disinfection (adding three The normal saline used for the normal saline is 0.9% NaCl solution, and the three antibodies added are penicillin, streptomycin and gentamicin. After adding the normal saline, penicillin, streptomycin and gentamicin The concentrations of mycin are 0.1 μl / mL, 0.1 μl / mL and 0.1 μl / mL respectively), and the soaking and disinfection time is 3 to 4 hours;
[0049] Removal of the spongy layer: In a sterile room, bluntly peel off the human amniotic membrane attached to the pretrea...
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Abstract
The invention discloses a dermis equivalent constructing method. The dermis equivalent constructing method comprises the following steps of: firstly, conducting sterile treatment on a human amniotic membrane, preparing the treated human amniotic membrane and a nitrocellulose membrane into a human amniotic membrane support; taking a milk goat ear skin specimen, removing dirty matters, disinfecting and washing the milk goat ear skin specimen, digesting a resultant dermis layer after the processing with 0.25% of pancreatin, filtering and collecting cells, and culturing the collected cells, thus obtaining dermal fibroblasts; mixing bovine type I collagen solution and DMEM (Dulbecco's Modified Eagle's Medium) solution uniformly, adjusting the pH value of the mixed solution by using NaOH solution, adding the dermal fibroblasts into the mixed solution, and mixing uniformly, thus obtaining collagen cell mixture solution; dropping the obtained collagen cell mixture solution on the human amniotic membrane support, coagulating, and then adding the collagen cell mixture solution in culture solution, culturing in an incubator, and then obtaining a dermis equivalent after the culture. According to the dermis equivalent constructing method of the invention, the human amniotic membrane is used as the support, when being transplanted, the dermis equivalent can be prevented from tearing, the operation operability is improved, after an operation, the human amniotic membrane and organism tissues are compatible, and at the initial stage of the transplanting, certain nutrition ingredients of the organisms can be absorbed, so that the healing of the skin can be promoted.
Description
technical field [0001] The invention belongs to the technical field of biological tissue engineering, and in particular relates to a method for constructing a dermis equivalent by using human amniotic membrane and collagen gel as a scaffold, that is, a method for constructing a dermis equivalent. Background technique [0002] In 1975, Rheinwald and Green invented the epithelial culture technology, which made the synthesis of artificial epidermis possible. However, the artificial epidermis can only be used for epidermal injuries and epidermis and superficial dermis injuries. When the full-thickness or near-full-thickness dermis is damaged, it is easy to form wound contractures, scars or open wounds. Therefore, the research and development of dermal substitutes has become a clinical demand. [0003] At present, artificial synthetic dermis mainly uses collagen-aminodextran, collagen gel, polyglycolic acid, nylon mesh, etc. as dermal scaffolds. For example, the outer layer of ...
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Patent Type & Authority Patents(China)
IPC IPC(8): A61L27/60A61L27/36A61L27/24
Inventor 史明艳高雪杨学义华进联易力任红艳张路陪高会江李俊雅许尚忠窦忠英
Owner INST OF ANIMAL SCI OF CHINESE ACAD OF AGRI SCI
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