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A method for preparing decellularized nerve graft by supercritical fluid extraction reaction

A supercritical fluid and decellularization technology, applied in the field of biomedical engineering, can solve the problems of long chemical decellularization reaction time, insufficient protein removal, poor penetrating power of water system, etc., and achieve good clinical use effect and small structural damage , the effect of short acting time

Active Publication Date: 2022-02-25
NANTONG UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In short, the currently widely used chemical decellularization system has disadvantages: 1. The concentration of the chemical decellularization reaction solution is strong and the reaction ability is strong; 2. The chemical decellularization reaction time is too long and the number of reactions is too many; Poor, protein removal is not thorough enough, high residual value

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] (1) Cut out 5 rat sciatic nerves, remove the extraneural fat, blood vessels and connective tissue, then cut them into 20mm lengths, and place them in distilled water for 12 hours in a shaking bath;

[0027] (2) Immerse the 5 sciatic nerves after oscillating bathing in 100mL mass concentration of 3% TritonX-200 solution, and place them in a 1000mL high-temperature and high-pressure reactor, and feed CO 2 The air in the kettle was discharged, and then at room temperature (25°C), the pressure was increased to a set pressure of 15Mpa, and the reaction was carried out for 1 hour. After the reaction was over, the reaction kettle was opened, and 5 sciatic nerves from which the immunogenic protein had been completely removed were taken out.

[0028] (3) Wash 5 sciatic nerves with 1% sodium lauryl sulfate solution with a mass concentration of 200 mL for 3 hours, then soak them in 200 mL of distilled water, replace the distilled water every 20 minutes, and finally obtain the pres...

Embodiment 2

[0030] (1) Cut out 5 rat sciatic nerves, remove the extraneural fat, blood vessels and connective tissue, then cut them into 20mm lengths, and place them in distilled water for 12 hours in a shaking bath;

[0031] (2) Immerse the 5 sciatic nerves after oscillating bathing in 1% TritonX-200 solution with a mass concentration of 100mL, and place them in a 1000mL high-temperature and high-pressure reactor, and feed CO 2 The air in the kettle was discharged, and then at room temperature (35° C.), the pressure was increased to a set pressure of 20 MPa, and the reaction was carried out for 3 hours. After the reaction was over, the reaction kettle was opened, and 5 sciatic nerves from which the immunogenic protein had been completely removed were taken out.

[0032] (3) Wash 5 sciatic nerves with 1% sodium lauryl sulfate solution with a mass concentration of 200 mL for 3 hours, then soak them in 200 mL of distilled water, replace the distilled water every 20 minutes, and finally obta...

Embodiment 3

[0034] (1) Cut out 5 canine sciatic nerves, remove the extraneural fat, blood vessels and connective tissue, then cut them into 30mm lengths, place them in distilled water and shake and soak them for 12 hours;

[0035] (2) Immerse the 5 canine sciatic nerves after oscillating bathing in 3% TritonX-100 solution with a mass concentration of 300mL, place them in a 1000mL high-temperature and high-pressure reactor, and feed CO 2 The air in the kettle was discharged, and then at room temperature (35°C), the pressure was increased to a set pressure of 25Mpa, and the reaction was performed for 2 hours. After the reaction is over, open the reaction kettle, take out 5 canine sciatic nerves, then pour out the residual liquid in the high-temperature and high-pressure reaction kettle, and then add 300mL of TritonX-100 solution with a mass concentration of 3%. Sciatic nerve immersion, CO 2 Exhaust the air in the reaction kettle and react again, and finally obtain 5 canine sciatic nerves f...

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PUM

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Abstract

The invention relates to the field of biomedical engineering, and discloses a method for preparing decellularized nerve grafts by supercritical fluid extraction reaction, comprising: cutting out peripheral nerves, removing fat, blood vessels and connective tissues outside them, and then cleaning; The nerve is immersed in a surfactant solution, placed in a high-temperature and high-pressure reactor, and CO 2 The air in the reactor was exhausted, and then the reaction was carried out. After the reaction, the peripheral nerve was taken out; then the peripheral nerve was washed with sodium dodecyl sulfate solution, and finally soaked in distilled water. The method of the present invention greatly reduces the dosage and concentration of chemical reagents in the decellularization process, has low reaction temperature, short action time, and few reaction times, can effectively reduce immunogenicity, and has little damage to nerve structure, and not only maintains the Schwann cell substrate The lamellar tubular structure is complete, the nerve basement membrane tube is complete and unobstructed, and the laminin in axon regeneration is retained, which has a good clinical application effect and is worthy of clinical promotion.

Description

technical field [0001] The invention relates to the field of biomedical engineering, in particular to a method for preparing decellularized nerve grafts by supercritical fluid extraction reaction. Background technique [0002] Peripheral nerve defect refers to the disconnection of peripheral nerve trunk caused by trauma or surgery, which is a common clinical disease. Nerve defects are divided into four degrees, among which the defects of degrees I and II can be overcome by changing the joint position or by pathological methods such as free nerve trunks, prepositioning, rerouting, lengthening or shortening of bone joints, and more than degrees III (or the distance of the defect is greater than that of the defect). Nerve diameter more than 4 times) defects must rely on nerve grafts or various substitute bridges to repair. So far, the most effective nerve transplantation is autologous nerve transplantation, which is to transplant the patient's own relatively unimportant nerves...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61L27/36A61L27/50
CPCA61L27/3604A61L27/3675A61L27/3687A61L27/3691A61L27/50A61L2430/32A61L2430/40
Inventor 杨宇民赵亚红张鲁中李贵才凌珏杨鹏翔
Owner NANTONG UNIVERSITY
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