Decellularized nerve scaffold and preparation method thereof

A decellularized and neural technology, applied in the field of decellularized neural scaffolds and preparation of decellularized neural scaffolds, can solve the problems of incomplete demyelination and weak destructive effect

Inactive Publication Date: 2019-06-21
王伟 +1
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In the Hudson method, anionic extractant Triton X-200 and amphoteric extractant SB-10 and SB-16 are used. During the process of decellularization of coarse nerves, there is mainly incomplete demyelination. Regarding this point, after previous research and discussion, we It is believed that it may be related to the following reasons: 1. The layered myelin sheath structure of the thick nerve plate is thicker and denser than that of small animal nerves; 2. The above-mentioned extractant has mild properties and weak destructive effect

Method used

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  • Decellularized nerve scaffold and preparation method thereof
  • Decellularized nerve scaffold and preparation method thereof
  • Decellularized nerve scaffold and preparation method thereof

Examples

Experimental program
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Embodiment

[0040] Take the nerve tissue (length 45mm, diameter 4mm) of the discarded limb after amputation in clinical operation, such as figure 1 As indicated, the extraneural blood vessels, fat and connective tissue were removed, then placed in liquid nitrogen for 1-3 minutes, placed at 25-37°C for 3-10 minutes, repeated freezing and thawing 3-4 times, and then placed at 25-37°C After hypotonic treatment for 3 to 6 hours, use sterilized distilled water as the hypotonic solution, place the isolated nerve after hypotonic treatment in 100 mL of PBS buffer containing 125 mmol / L SB-10, and place it at 20 to 25 °C at 80 ~100 times / min frequency vibration soaking for 12~18h, then placed in 100mL mixed solution containing 0.14% Triton X-200 and 0.6mmol / L SB-16, at 20~25℃, at 80~100 times / min Soak for 9-12 hours by shaking at a frequency of 9-12 hours, then wash with sterilized PBS buffer, place the treated isolated nerve tissue in 100mL isotonic solution Ⅰ containing 125mmol / L SB-10, at 10-15°...

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Abstract

The invention relates to a preparation method of a decellularized nerve scaffold. The preparation method includes following steps: taking and pretreating isolated nerve tissue; repeatedly freeze-thawing the pretreated isolated nerve tissue; hypotonically treating isolated nerve; using a solution containing a detergent to treat the isolated nerve to remove cells, using nuclease to treat the isolated nerve, and cleaning to obtain the decellularized nerve scaffold. By using the method, nerve basal membrane tube and neural substrate structure are retained to greatest extent, and antigenic ingredients like lipid, protein and nucleic acid are removed sufficiently.

Description

technical field [0001] The invention relates to the field of tissue engineering materials, in particular to a method for preparing decellularized nerve scaffolds and the decellularized nerve scaffolds prepared by the method. Background technique [0002] Clinically, nerve injury caused by various reasons is very common. When the defect is large (>3cm), it is usually difficult to perform tension-free anastomosis, and nerve transplantation is required. Among them, although autologous nerve transplantation has a certain curative effect and is used as the gold standard for comparison with other treatment methods, there are limitations such as limited sources and ethical issues, which limit its further use. However, the current level of technology is still unable to imitate artificial scaffolds similar to the structure and function of natural extracellular matrix, while the natural structure inside the decellularized allogeneic nerve is similar to the extracellular matrix stru...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/36A61L27/50
Inventor 王伟李杰马义勇智晓东李宇张玉强张振宇管树军邱裕佳段思腾张臣鸣曹宇
Owner 王伟
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