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Artificial nerve graft and preparation method based on sliced ​​decellularized scaffold

A technology of nerve transplantation and grafting, which is applied in the direction of animal cells, vertebrate cells, bone/connective tissue cells, etc., can solve the problems of low decellularization strength, residual, and influence on cytokine activity, etc., and achieve the improvement of cell loading capacity and promote Proliferation of stem cells, effect that contributes to cell attachment

Active Publication Date: 2020-09-25
JIANGSU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The technical defects in the current technology are as follows: (1) Decellularization of the whole tissue has too little decellularization strength, which is easy to remain, and if the strength is too high, it will be crushed; (2) Planting seed cells on the whole tissue must rely on the seed cells to penetrate the tissue Invasion and migration, otherwise the cell-carrying capacity of the scaffold with only the presence of cells on the surface of the scaffold is low; (3) Similarly, the overall tissue cross-linking cytokines must also rely on factors to penetrate into the tissue, and there are many influencing factors
[0005] At present, the commonly used cross-linking agents for cross-linking cytokines to scaffolds include glutaraldehyde, peroxidase, polyphenol oxidase and carbodiimide, etc. Among them, glutaraldehyde has cytotoxicity, and peroxidase and polyphenol oxidase Can significantly affect the function of cytokines, carbodiimide is not only expensive but also affects the activity of cytokines

Method used

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  • Artificial nerve graft and preparation method based on sliced ​​decellularized scaffold
  • Artificial nerve graft and preparation method based on sliced ​​decellularized scaffold

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Embodiment Construction

[0030] Further illustrate the present invention below in conjunction with specific embodiment:

[0031] Rats (400-500 g Sprague Dawley rats; from the Experimental Animal Center of Jiangsu University) were deeply anesthetized with chloral hydrate, and after about 500 mL of phosphate buffer solution was perfused through the heart, 4% (wt.%) paraformaldehyde 500 mL, fully fixed rat tissue. The rat spinal canal was dissected, the spinal cord was carefully separated, the meninges on the surface were carefully removed, and then fixed in 4% paraformaldehyde for about 2 hours. Embedded in OCT embedding medium, the Leica cryostat was used to cut longitudinally at -15°C, and the slice thickness was selected as 60 μm. After the slices were taken out, they were preliminarily washed in phosphate buffer, and then entered into the decellularization program.

[0032] Soak slices in 3% (wt.%) Triton X-100 for 2-3 h, then wash with a large amount of phosphate buffer; wash after soaking in 1M ...

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Abstract

The invention relates to an artificial nerve graft constructed based on a chip-type acellularized scaffold and a preparation method of the artificial nerve graft, belonging to the field of tissue engineering. According to the artificial nerve graft and the preparation method, heterogeneous animal medulla spinalis, skeletal muscle or tendon acellularized tissue slices are adopted as a bracket, genipin is adopted for crosslinking cytokines, autologous ecto mesenchymal stem cells are planted on the bracket, and fibrous protein is taken as an adhesive for constructing the artificial nerve graft, so that the repairing of the nervous tissue is promoted. The artificial nerve graft and the preparation method have the advantages that the preparation cost is low, the period is short, the preparation raw materials are easily available, and the preparation is simple and is easy to operate. The constructed artificial nerve graft has low immunogenicity, contains a large number of autologous stem cells and cell factors, and is beneficial for adjusting the nerve injury microenvironment, and promoting the nerve regeneration and myelinogenesis. The artificial nerve graft can be used for preparing peripheral nerve and medulla spinalis defect, and has the broad application prospect.

Description

technical field [0001] The invention belongs to the field of tissue engineering, and in particular relates to an artificial nerve graft constructed on the basis of sliced ​​decellularized scaffolds and a preparation method. Background technique [0002] Defects are often caused after nerve injury, especially after spinal cord injury will cause cavities. How to fill the defect and reduce the formation of cavities is a hot issue in the field of nerve injury repair. Although autologous nerve transplantation can treat nerve defects, obvious side effects and limited sources severely limit its application. The focus of current research is to use the concept of tissue engineering to construct nerve grafts to fill the defect and promote the repair of nerve damage. [0003] Scaffolds, cytokines and seed cells are the three key elements of tissue engineering. Although artificial polymer materials such as polylactic acid have a wide range of sources, they have disadvantages such as u...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/077A61L27/54A61L27/52A61L27/50A61L27/36A61L27/38A61L27/22
CPCA61L27/225A61L27/3604A61L27/3675A61L27/3687A61L27/3834A61L27/3878A61L27/50A61L27/52A61L27/54A61L2300/414A61L2430/32C08L89/00
Inventor 陈谦胡萍萍陈平波龚爱华张志坚
Owner JIANGSU UNIV
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