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Tissue engineered skin with basilar membrane and construction method thereof
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A technology of tissue engineering skin and construction method, which is applied in the field of burn and wound skin defect repair and scar plastic surgery, to achieve the effect of mature and firm development, simple production method, and convenient material extraction
Inactive Publication Date: 2014-06-04
SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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Problems solved by technology
[0004] There is no literature report at home and abroad to use amniotic membrane as basement membrane and composite plasma as matrix to construct tissue-engineered skin with basement membrane through in vitro culture.
Method used
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Examples
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Embodiment 1
[0024] Example 1. Preparation of Active Skin Substitutes Containing Basement Membrane
[0025] 1. Isolation and culture of human keratinocytes and fibroblasts
[0026] The discarded foreskin tissue after circumcision was used to separate and culture keratinocytes and fibroblasts by enzymatic digestion. Separately prepared into a single cell suspension, according to 2-3 × 10 5 Cells per ml were seeded in culture flasks and placed in a 37°C incubator. Keratinocytes were cultured in serum-free medium (DK-SFM, Gibco, USA), and fibroblasts were cultured in complete DMEM medium (Gibco, USA). USA) for culture, when the cells reached 70%-80% confluence, passage and expansion were carried out.
[0028] Strip off the fresh amniotic membrane under aseptic conditions (taken from the placental tissue of caesarean section women who are negative for hepatitis virus antibody, syphilis antibody and HIV), bluntly separate the chorion, s...
Embodiment 2
[0031] Embodiment 2. Animal transplantation test of tissue engineered skin of the present invention
[0032] Male Balb / c-nu mice (nude mice, provided by Shanghai Xipuer-Bikay Experimental Animal Co., Ltd.), body weight 18 ± 2 grams, 16, were divided into two groups, respectively, the tissue engineered skin containing amniotic membrane of the present invention Group A and the traditional active skin substitute without amniotic membrane Group B: purely constructed with fibrin as a matrix (see A.L.Mazlyzama, B.S.Aminuddinb, et al.Reconstruction of living bilayer human skin equivalent utilizing human fibrin as a scaffold.Burns , 2007, 33(3): 355-363).
[0033]After the nude mice were anesthetized with ketamine intraperitoneally, the skin and hair on the back were shaved, and the full-thickness skin on the ventral side of the spine and the deep fascia up to the muscle layer were excised. Group A: The tissue-engineered skin cultured in vitro for 2-3 weeks described in Example 1 was...
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Abstract
The invention relates to the technical fields of tissue engineering and medical wound repair. At present, a living skin substitute constructed by using the materials of polylactic acid, polyglycolic acid, collagen, hyaluronic acid, and the like as a dermic bracket has the defects that on one hand, host materials are difficult to extract, the living skin substitute has complicated manufacturing technology and is expensive in cost and difficult to widely popularize and apply clinically; and on the other hand, the living skin substitute does not have a skin basilar membrane structure so that healed skin does not resist pressure and wear, and the living skin has unfirm adhesion to the epidermis and is easy to shed and break or form water blisters so that the structural and morphological development of the normal epidermis is influenced; and allogeneic acellular dermis is taken from cadaver skin and is limited in sources and expensive in cost, and thus clinical application is limited. The invention aims at providing a skin substitute which uses surface-finished and modified amnion as the basilar membrane and blood plasma as stroma, which has the advantages of wide material sources, low cost and simple preparation method. An animal experiment proves that the complete basilar membrane and hemidesmosomes can be retained in in-vivo transplantation, and the formation of an epidermal structural form is accelerated and promoted.
Description
technical field [0001] The invention relates to the technical field of tissue engineering and medical wound repair, in particular to a tissue-engineered skin (i.e., an active skin substitute) in which the surface of amniotic membrane is modified and modified as a basement membrane, and the composite plasma is used as a matrix, which is cultured in vitro to construct a tissue-engineered skin containing a basement membrane. , especially suitable for skin defect repair of burns and trauma and scar shaping. Background technique [0002] At present, biodegradable materials such as polylactic acid, polyglycolic acid, collagen, and hyaluronic acid are used to construct dermal scaffolds, and active skin substitutes are constructed on this basis, which have been initially applied to the repair of deep skin defect wounds and scar plastic surgery. However, on the one hand, it is difficult to extract the matrix material, the manufacturing process is complicated, and the cost of the skin...
Claims
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Application Information
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Patent Type & Authority Patents(China)
IPC IPC(8): A61L27/60C12N5/071
Inventor 纪世召夏照帆肖仕初李恒宇王光毅杨晓妍罗鹏飞
Owner SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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