Epidermal graft as well as preparation method and application thereof

A kind of epidermis and skin technology, applied in the field of biological products, can solve the problems of limited self-proliferation ability, difficult to obtain, and easy aging of seed cells, so as to facilitate cell migration and proliferation and infiltration of growth factors, promote repair, regeneration and healing, and promote healing effect

Inactive Publication Date: 2016-08-10
GENERAL HOSPITAL OF PLA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

With the further deepening of research on large-area wounds, tissue-engineered skin has gradually developed in the application of research on large-area wounds, but one of the difficulties (as mentioned above) is the seed cells currently used to construct tissue-engineered skin (such as Keratinocytes and fibroblasts) have limited self-proliferation ability, are easy to age, are not easy to obtain, and have certain immune rejection, which leads to certain limitations in the use of tissue engineering technology to treat large-area wounds; the second difficulty is that the mesenchymal Mesenchymal stem cells are used in the repair and regeneration of large-scale wounds, and the problem of how to use mesenchymal stem cells to obtain tissue-engineered skin safely, stably, quickly and with high quality is becoming more and more prominent, and it is a problem that needs to be solved urgently

Method used

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  • Epidermal graft as well as preparation method and application thereof
  • Epidermal graft as well as preparation method and application thereof
  • Epidermal graft as well as preparation method and application thereof

Examples

Experimental program
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preparation example Construction

[0024] In a first aspect, the present invention provides a method for preparing an epidermal sheet, the method comprising: preparing a collagen-chitosan porous scaffold, then seeding mesenchymal stem cells on the collagen-chitosan porous scaffold, and then injecting the collagen-chitosan porous scaffold. Induction and culture of mesenchymal stem cells on chitosan porous scaffolds into epidermal cells.

[0025] In the preparation method of the present invention, the method for preparing the collagen-chitosan porous scaffold is not particularly limited, and may be various preparation methods commonly used in the art. But, the inventor of the present invention finds in research, when preparing collagen-chitosan porous support, control the specific concentration of collagen and chitosan and the pH value of collagen-chitosan solution in collagen-chitosan solution Collagen-chitosan porous scaffolds with different pore sizes will be obtained. Therefore, in order to further seek a co...

Embodiment

[0049] The following examples will further illustrate the present invention, but do not limit the present invention thereby.

[0050] The experimental methods in the following examples and comparative examples, unless otherwise specified, are conventional methods in the art. The experimental materials used in the following examples and comparative examples are commercially available unless otherwise specified.

[0051] Chitosan powder was purchased from Sigma Company.

[0052] The scanning electron microscope was purchased from Hitachi, Japan, model TM100.

[0053] SD rat bone marrow mesenchymal stem cells were purchased from Tianjin Weikai Bioengineering Co., Ltd.

[0054] The preparation method of EDC / NHS cross-linking agent is: dissolve EDC, NHS respectively in 95% ethanol solution to concentration to be 33nmol / L and 8nmol / L, obtain EDC solution and NHS solution, then equal volume of EDC solution and NHS solution mix.

preparation example

[0056] This preparation example is used to illustrate the extraction method of collagen powder.

[0057] (1) Take 10 g of fresh bovine tendon, remove the surrounding connective tissue, and cut the bovine tendon into 1-2mm sized tissue pieces.

[0058] (2) Put the tissue block into 0.1 mass% Na 2 CO 3 Soak in the solution for 2h, rinse with distilled water 3 times, 10min each time.

[0059] (3) Soak the tissue block in 0.5% Tris-HCl (pH=7.5) solution containing 1 mol / L NaCl for 12 hours, rinse with distilled water for 3 times, 10 minutes each time.

[0060] (4) Put the tissue pieces into 250 mL of 0.05 mol / L acetic acid solution containing 500 mg of pepsin, soak at 4°C for 72 hours, and stir repeatedly until fully digested into a transparent paste.

[0061] (5) The mixture was centrifuged at 5000 g for 20 min at 4° C., and the supernatant was taken.

[0062] (6) Perform the operations of steps (4) and (5) on the centrifuged sediment in step (5), and repeat the operation 2 t...

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Abstract

The invention relates to the field of biological products, and discloses epidermal graft as well as a preparation method and application thereof. The preparation method disclosed by the invention comprises the following steps: preparing a collagen-chitosan porous bracket, inoculating mesenchymal stem cells to the collagen-chitosan porous bracket, and performing induction culture on the mesenchymal stem cells on the collagen-chitosan porous bracket, so as to obtain epithelial cells. By adopting the epidermal graft, regeneration, reconstruction, remediation and heal of a large-area wound skin surface can be remarkably, efficiently and safely promoted.

Description

technical field [0001] The invention relates to the field of biological products, in particular to an epidermal sheet, its preparation method and its application in the preparation of skin substitutes. Background technique [0002] For large full-thickness skin defects, autologous or allogeneic skin grafting is commonly used clinically to repair them, but these methods will cause new trauma to the donor site. At the same time, the source of donor skin and immune rejection also limit its effective application. With the development of tissue engineering technology, tissue engineered skin has become the first tissue engineering product used clinically. Although these products can temporarily cover the wound, there are still certain immune rejection reactions, and the therapeutic effect is not ideal. The key issues of tissue engineering are seed cells and scaffold structure. At present, the commonly used seed cells are autologous or allogeneic keratinocytes and fibroblasts, bu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/60A61L27/56A61L27/50A61L27/38A61L27/20A61L27/24
CPCA61L27/20A61L27/24A61L27/3834A61L27/50A61L27/56A61L27/60C08L89/00C08L5/08
Inventor 韩为东付小兵郝好杰陈德云郑敬曦
Owner GENERAL HOSPITAL OF PLA
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