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Application of acellular nerve hydrogel to preparation of peripheral nerve injury repairing composition

A peripheral nerve and decellularization technology, applied in the field of biomedical composite materials, can solve problems affecting nerve sensory and motor functions, and achieve the effect of repairing peripheral nerve damage

Inactive Publication Date: 2017-05-31
GUANGZHOU SUN SHING BIOTECH CO LTD +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the hydrogel with various neurotrophic factors can promote the growth of axons, it also promotes the excessive branching of neurons and the formation of synapses between neurites. For the central nervous system, the formation of synapses between neurons It is a necessary condition for the normal function of the nerve tissue, but for the peripheral nerve, if the nerve fibers form a large number of synaptic connections with each other during the process of walking, it will cause pathological neuralgia and neuroma, which will affect the normal feeling and nerve function of the nerve. Motor function, how to optimize the biological function of the hydrogel to adapt to the needs of peripheral nerve regeneration will be the future research direction

Method used

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  • Application of acellular nerve hydrogel to preparation of peripheral nerve injury repairing composition
  • Application of acellular nerve hydrogel to preparation of peripheral nerve injury repairing composition
  • Application of acellular nerve hydrogel to preparation of peripheral nerve injury repairing composition

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Embodiment 1: the preparation of hydrogel

[0036] The preparation method of decellularized nerve hydrogel is as follows:

[0037] 1) Take the peripheral nerve of the rat, cut off the fat tissue and epineurium on the surface under an operating microscope, shake the treated nerve in distilled water, and rinse it for 6 hours;

[0038] 2) Extraction: Put the nerve in 3% Triton X100 aqueous solution and shake for 12 hours, then rinse it in distilled water for 3 times, then put it in 4% sodium deoxycholate aqueous solution and shake it at room temperature for 24 hours, and finally rinse it in distilled water for 3 times ; So cycle twice the extraction process;

[0039] 3) Freeze-drying: put the extracted nerve into a mixed solution of ethanol and dichloromethane to degrease, wherein the volume ratio of ethanol to dichloromethane is 1:2, then freeze-dry and grind into powder;

[0040] 4) Dissolution and digestion: Add the lyophilized powder to hydrochloric acid solution to ...

Embodiment 2

[0043] Example 2: Effect test of decellularized nerve hydrogel on axon growth

[0044] The DRG tissues of rats born one day old were taken out, respectively planted in 6-well plates pre-coated with decellularized nerve hydrogel (prepared in Example 1), type I collagen, and two different hydrogels, and cultured at 37°C for 48h After the PH microscope was observed and photographed, the experimental results were as follows figure 1 shown.

[0045] In the figure, A indicates that the neurites can be seen to grow radially around the DRG tissue block on the culture plate coated with decellularized nerve hydrogel, and the inner circle is drawn with the boundary of the tissue block as a reference, and the neurite growth can reach the farthest Draw concentric outer circles for the radius. B: Partial enlarged view showing the growth of neurites and cell migration, and the distance to the furthest distance of neurites was measured from the border of DRG. C and D show neurites and axon...

Embodiment 3

[0046] Example 3: Effect test of decellularized nerve hydrogel on arrangement of Schwann cells

[0047] The DRG tissue block was digested into a single cell suspension (mainly including DRG neurons and Schwann cells), and planted on decellularized nerve hydrogel (prepared in Example 1), Matrigel (#356234, BD) and type I collagen In 24-well plates pretreated with three different hydrogels, observed after 2 days of adherent culture + 7 days of myelinating culture. The result is as figure 2 shown.

[0048] It is known from the figure that the decellularized neural hydrogel environment can induce spindle cells (via image 3Proved to be Schwann cells) linearly arranged in a bead-like structure (marked by the arrow), and bare neurites are rarely seen in the field of view. On the culture plate coated with Matrigel and type Ⅰ collagen, the arrangement of spindle cells was disordered, and a large number of exposed neurites could be seen at the same time. The results show that the ...

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Abstract

The invention provides application of acellular nerve hydrogel to preparation of a peripheral nerve injury repairing composition. The peripheral nerve injury repairing composition is prepared by using the acellular nerve hydrogel, so the peripheral nerve regeneration microenvironment can be simulated well, extension and myelination of axon can be promoted, the PSD95 expression level of nerve cells is reduced, and occurrence of pathological neuralgia and neuroma is inhibited. An electrophysiological experiment proves that although the acellular nerve hydrogel inhibits formation of cynapse between the nerve cells, the acellular nerve hydrogel does not influence generation of action potential and conduction along axion and can repair peripheral nerve injury well.

Description

technical field [0001] The invention belongs to the field of biomedical composite materials, and in particular relates to the use of decellularized nerve hydrogel for preparing peripheral nerve injury repair compositions Background technique [0002] Biohydrogels prepared from chitosan, collagen and other biomaterials not only have good biocompatibility in chemical properties, but also are similar to extracellular matrix in physical properties, so they are widely used in wound dressings, tissue Engineering supports and other fields. Loading various neurotrophic factors in the hydrogel can further promote the growth of nerves. [0003] CN102657874A discloses a hydrogel-wrapped lentivirus Lingo-1 RNAi complex for repairing nerve damage and a preparation method thereof. This document discloses the strategy of using Lingo-1 RNAi combined with hydrogel bio-scaffold and release. RNAi interferes with Lingo-1 and cooperates with hydrogel to promote axon regeneration in spinal cord...

Claims

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Application Information

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IPC IPC(8): A61L27/36A61L27/52A61L27/56
CPCA61L27/52A61L27/3633A61L27/3675A61L27/56A61L2400/06A61L2430/32
Inventor 邹剑龙曾圆山全大萍刘晟孙佳慧杨伟红刘小林朱庆棠邱帅曾晨光杨习锋
Owner GUANGZHOU SUN SHING BIOTECH CO LTD
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