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Mesenchymal stem cell tissue engineering scaffold and preparation method and application thereof

A tissue engineering scaffold and stromal cell technology, applied in the field of mesenchymal stem cell tissue engineering scaffold and its preparation, can solve problems such as dysfunction, lack of effective means for mesenchymal stem cells, cell apoptosis, etc. The effect of cell proliferation and migration and inhibition of inflammatory response

Inactive Publication Date: 2016-08-03
GENERAL HOSPITAL OF PLA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

With the deepening of research on difficult-to-heal wounds, although the unique role of mesenchymal stem cells has gradually developed in the application of research on difficult-to-heal wounds, one of the difficulties today is the application of mesenchymal stem cells in wound repair and regeneration. In the absence of effective means, it can be found in previous studies that due to the harsh environment of chronic small pieces that do not heal the wound, a large number of cells injected intravenously or locally cannot migrate to the wound, and there are still problems of apoptosis and dysfunction, and how to safely The problem of stable, rapid and high-quality use of mesenchymal stem cells is becoming more and more prominent, and it is a problem that needs to be solved urgently

Method used

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  • Mesenchymal stem cell tissue engineering scaffold and preparation method and application thereof
  • Mesenchymal stem cell tissue engineering scaffold and preparation method and application thereof
  • Mesenchymal stem cell tissue engineering scaffold and preparation method and application thereof

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Experimental program
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preparation example Construction

[0022] In a first aspect, the present invention provides a method for preparing a tissue engineering scaffold of mesenchymal stem cells, the method comprising: preparing a gelatin-chitosan porous scaffold, and then inoculating mesenchymal stem cells to the gelatin-chitosan porous scaffold, and incubated.

[0023] In the preparation method of the present invention, the method for preparing the gelatin-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 gelatin-chitosan porous support, control the specific concentration of gelatin and chitosan in gelatin-chitosan solution and the pH value of gelatin-chitosan solution Gelatin-chitosan porous scaffolds with different pore sizes will be obtained. Therefore, in order to further seek a gelatin-chitosan porous scaffold more suitable for the growth of mesenchymal stem cells, preferably, the me...

Embodiment

[0048] The following examples will further illustrate the present invention, but do not limit the present invention thereby.

[0049] 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.

[0050] Chitosan powder was purchased from Sigma Company.

[0051] The scanning electron microscope was purchased from Hitachi, Japan, model TM100.

[0052] SD rat bone marrow mesenchymal stem cells were purchased from Tianjin Weikai Bioengineering Co., Ltd.

[0053] 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

[0055] This preparation example is used to illustrate the extraction method of gelatin powder.

[0056] (1) Take 10 g of fresh bovine tendon, remove the surrounding connective tissue, and cut the bovine tendon into 1-2mm sized tissue pieces.

[0057] (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.

[0058] (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.

[0059] (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.

[0060] (5) The mixture was centrifuged at 5000 g for 20 min at 4° C., and the supernatant was taken.

[0061] (6) Perform the operations of steps (4) and (5) on the centrifuged sediment in step (5), and repeat the operation 2 ti...

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Abstract

The invention relates to the field of biological products and discloses a mesenchymal stem cell tissue engineering scaffold, a preparation method of the scaffold and application of the scaffold in skin substitutes. The preparation method of the scaffold comprises the steps of preparing a gelatin-chitosan porous scaffold and then inoculating the mesenchymal stem cells to the gelatin-chitosan porous scaffold for incubation and cultivation. The mesenchymal stem cell tissue engineering scaffold can obviously and effectively promote regeneration, reconstruction, repair and healing of skin wound surfaces difficult to heal, such as diabetes.

Description

technical field [0001] The invention relates to the field of biological products, in particular to a mesenchymal stem cell tissue engineering scaffold and its preparation method and application. Background technique [0002] Because the tissue cells of diabetic patients are in a high-glucose environment for a long time, the local skin tissue cells and proteins are glycosylated, causing abnormal vascular endothelial function and impaired response; the function of macrophages is weakened, and the inflammatory response is prolonged and unbalanced; growth The lack or damage of factors, the reduction of gelatin tissue, the disorder of epidermal barrier function, and the reduction of granulation tissue make the ulcer difficult to repair. Among them, 15% of diabetic patients have foot ulcers, a small piece of chronic wound that has not healed, which seriously affects the quality of life of patients. And the research on the treatment of refractory skin wounds caused by diabetes has...

Claims

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Application Information

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IPC IPC(8): A61L27/20A61L27/22A61L27/38A61L27/50A61L27/56A61L27/60
CPCA61L27/20A61L27/222A61L27/3834A61L27/50A61L27/56A61L27/60C08L89/00C08L5/08
Inventor 韩为东付小兵郝好杰佟川郑敬曦
Owner GENERAL HOSPITAL OF PLA
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