Nerve scaffold for nerve injury repairing and preparing method and application thereof

A nerve injury and nerve technology, applied in the field of nerve regeneration biology, can solve the problems of low utilization rate, lack, and low survival rate of neural stem cells, and achieve the effect of remarkable repair effect, simple preparation process and simple production process.

Inactive Publication Date: 2016-05-25
SHANGHAI SHENYIN BIOTECH
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] In nerve regeneration engineering, the neural stem cells transplanted into the host can chemotaxis, migrate and gather to the lesion of the nervous system, and can survive, proliferate and differentiate into neurons and glial cells, thereby promoting the recovery of the patient's missing function. However, previous studies have shown that the survival rate of neural stem cells after allogeneic transplantation is very low due to immune reactions, etc., so the utilization rate of neural stem cells is low

Method used

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  • Nerve scaffold for nerve injury repairing and preparing method and application thereof
  • Nerve scaffold for nerve injury repairing and preparing method and application thereof
  • Nerve scaffold for nerve injury repairing and preparing method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] 1) Purchase 180-220g rats, kill the rats by cardiac perfusion, cut off the tail, extract the silver tail tendon from the tail, put it in deionized water, put it on a shaker, temperature 4°C, speed 50-200rpm , repeated washing 5 times, the washed tail tendon was dissolved in 0.05% to 0.1% glacial acetic acid-PBS at a temperature of 4°C, left standing for 2 to 5 days, and the collagen concentration was adjusted to 10mg / ml.

[0052] 2) Take the upper layer of collagen solution, put it into a mold, put the mold into liquid nitrogen to freeze, and then put it in a 2.5L&6L freeze dryer for 8-12 hours to freeze-dry to obtain a collagen sponge, which is a three-dimensional collagen material with a pore size of 30-200 μm , the morphology of the collagen sponge see figure 1 .

[0053] 3) Utilize electron beam irradiation to sterilize the collagen sponge, the treatment condition is: 12kGy 60 Co.

[0054] 4) Dissolve heparin in the cross-linking solution containing 4mMEDC and 2....

Embodiment 2

[0059] 1. The preparation method of the nerve conduit that is used for repairing nerve damage, comprises the steps:

[0060] 1) Purchase 180-220g rats, kill the rats by cardiac perfusion, cut off the tail, extract the silver tail tendon from the tail, put it in deionized water, put it on a shaker, temperature 4°C, speed 50-200rpm , repeated washing 5 times, the washed tail tendon was dissolved in 0.05% to 0.1% glacial acetic acid-PBS at a temperature of 4°C, left standing for 2 to 5 days, and the collagen concentration was adjusted to 10mg / ml.

[0061] 2) Take the upper layer of collagen solution, put it into a tubular mold, put the mold into liquid nitrogen to freeze, and then put it in a 2.5L&6L freeze dryer for 8-12 hours to freeze-dry to obtain a hollow tubular collagen sponge, that is, a collagen tube.

[0062] Collagen sponge is the carrier of growth factors and neural stem cells, so the hollow part of the obtained collagen tube can be filled with some collagen sponges t...

Embodiment 3

[0077] 1. The difference between the preparation method of the nerve guide put in for nerve injury repair in this embodiment and the preparation method of the nerve guide in Example 2 is that the collagen concentration is 20 mg / ml, and the concentration of the cross-linking agent in the cross-linking solution is: 10 mMEDC and 5 mM NHS, the concentration of heparin is 4 mg / mL, the concentration of bFGF is 500 μg / mL, and the rest of the operation steps are the same as in Example 2.

[0078] 2. Detection of cell proliferation and activity by CCK-8.

[0079] Set up 3 groups of parallel experiments on neural scaffolds, the first group is the group of this embodiment, in the neural scaffolds of this group, bFGF is fixed on the collagen scaffold by heparin, and is cultured with neural stem cells (HS-bFGF group for short); For the control group, in the nerve scaffolds of this group, bFGF was not fixed with heparin, but the bFGF was simply incubated on the collagen scaffold, and neural...

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Abstract

The invention discloses a nerve scaffold for nerve injury repairing and a preparing method and application thereof. The nerve scaffold comprises a collagen scaffold body, neural stem cells cultured on the collagen scaffold body and growth factors fixed to the collagen scaffold body through heparin. The growth factors have heparin binding domains. The preparing method includes the steps that a collagen solution is subjected to freeze drying before crosslinking, a collagen sponge obtained after freeze drying is soaked into a crosslinking solution to be crosslinked, the growth factors are fixed to the collagen scaffold body through the heparin, and the activity of the neural stem cells is maintained or propagation of the neural stem cells is promoted. According to the preparing method, materials are convenient to obtain, the steps are simple, complex steps including acid-base neutralization, dialysis and the like are avoided, the opportunity that collagen materials are in contact with harmful reagents is reduced, the safety of the obtained nerve scaffold is improved, the nerve scaffold can be molded to be in different shapes as required and can be used for filling nervous tissue losses and bridged nerve broken ends and guiding growth of nerves, nerve injury repairing and function recovery are promoted accordingly, and good stability is achieved.

Description

technical field [0001] The invention belongs to the field of nerve regeneration biology, and in particular relates to a nerve support for repairing nerve damage, its preparation method and application. Background technique [0002] Nervous system injuries mainly include spinal cord injury, brain injury, and peripheral nerve injury, with serious consequences, and their treatment has always been a research hotspot. The incidence of nervous system injury has been increasing year by year with the development of economy and transportation, which has attracted more and more attention from clinicians and scientific researchers. [0003] Once the nervous system is damaged, the degree of recovery is limited and the treatment effect is not good. For a long time, some nervous system diseases still have serious sequelae after traditional treatment. Therefore, choosing an exogenous substance to replace the already Thoroughly damaged tissue can promote the regeneration of residual nerve ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/24A61L27/36A61L27/54A61L27/20
CPCA61L27/20A61L27/24A61L27/3604A61L27/3675A61L27/54A61L2300/414A61L2430/32C08L89/00C08L5/10
Inventor 马富凯朱剑虹朱侗明王智富
Owner SHANGHAI SHENYIN BIOTECH
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