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An axial gradient protein-loaded silk nerve conduit and its preparation and application

A nerve conduit and silk fibroin technology, applied in the field of biomedical engineering, can solve problems affecting the effect of the action

Active Publication Date: 2021-09-10
NANTONG UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Studies have shown that nerve growth factor can promote regeneration in the early stage of nerve regeneration, and may inhibit regeneration in the later stage. However, the uncontrollable and unstable protein (nerve growth factor) load directly affects the effect of these growth factors in vivo

Method used

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  • An axial gradient protein-loaded silk nerve conduit and its preparation and application
  • An axial gradient protein-loaded silk nerve conduit and its preparation and application
  • An axial gradient protein-loaded silk nerve conduit and its preparation and application

Examples

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preparation example Construction

[0033] 2) Preparation of a photoactive silk nerve guide: dissolving the silk fibroin modified with o-nitrobenzyl photosensitive molecules obtained in step 1) in hexafluoroisopropanol, adding ultrapure water while stirring, Obtain the mixed solution and inject it into the mold, seal it away from light and let it stand for 1 to 5 days, then take it out and wash away the hexafluoroisopropanol in ultrapure water to obtain a photoactive silk nerve guide;

[0034] 3) Preparation of axially graded protein-loaded silk nerve conduits: soak the photoactive silk nerve conduits obtained in step 2) in a nerve growth factor solution, and use blue light to sequentially reduce the irradiation time along the axial direction on the The front region, the middle region and the end region of the photoactive silk nerve guide are irradiated, and then washed with PBS buffer solution under ultrasonic conditions and then freeze-dried to obtain an axially gradient protein-loaded silk nerve guide, wherein...

Embodiment 1

[0054] Preparation of O-Nitrobenzyl Photosensitive Molecules (ONB)

[0055] (1) Dissolve 8.9g of 4-hydroxy-3-methoxybenzaldehyde in 40mL of dimethylformamide (DMF) (concentration 0.2225g / mL), then add 9.89g (1.11 equivalents) of 4-bromobutyric acid methyl Esters and 10.2g (1.15 equivalents) of potassium carbonate were stirred and reacted at room temperature for 16 hours, and then the reaction solution was poured into ice water. After precipitation and suction filtration, the filter cake was dissolved in dichloromethane, and anhydrous magnesium sulfate was added to dry and remove water, and filtered Magnesium sulfate was removed, and the collected filtrate was evaporated to remove dichloromethane under reduced pressure to obtain the first intermediate product methyl4-(4-formyl-2-methoxyphenoxy)butanoate (molecular formula 1).

[0056] (2) Take 9.4g of the first intermediate product and slowly add it into 140mL of 65% nitric acid solution in v / v under ice-water bath conditions. ...

Embodiment 2

[0059] Preparation of silk fibroin:

[0060] (1) Heat 2000mL of sodium carbonate solution with a concentration of 0.02mol / L to 100°C, add 50g of mulberry silk and continue to boil for 30min, remove sericin, wash and soak in ultrapure water until the washing liquid becomes neutral.

[0061] (2) Dissolve 20g of mulberry silk from which sericin has been removed in 100mL lithium bromide solution with a concentration of 9.3mol / L at 60°C, react for 4h, cool to room temperature, dialyze with ultrapure water for 3 days, and then freeze-dry to obtain 15g silk fibroin.

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Abstract

The invention relates to the field of biomedical engineering, and discloses an axial gradient protein-loaded silk nerve guide and its preparation and application. In the present invention, by grafting o-nitrobenzyl photosensitive molecules on silk fibroin, silk fibroin modified by o-nitrobenzyl photosensitive molecules is obtained, and then prepared into a photoactive silk nerve guide, and finally The silk nerve guide is soaked in the nerve growth factor solution, the front end, the middle part and the end are irradiated with blue light along the axial direction, after washing and freeze-drying, an axial gradient protein-loaded silk nerve guide is obtained. The invention gradually reduces the light irradiation time in the front, middle and end regions of the silk nerve guide, and realizes the precise and controllable gradient loading of the silk nerve guide. The biological activity of the silk nerve guide of the invention is greatly improved, can effectively promote the proliferation and migration of nerve cells, can induce the rapid growth of new nerve tissue, and achieve good nerve repair effect in clinical practice.

Description

technical field [0001] The invention relates to the field of biomedical engineering, in particular to an axial gradient protein-loaded silk nerve guide and its preparation and application. Background technique [0002] Rapid and effective repair of peripheral nerve injury (PNI) is one of the unsolved worldwide clinical problems. Every year, there are about 2 million new PNI patients in the world, so there is a huge clinical demand for PNI repair. However, various tissue-engineered nerve bridges (stents / catheters) currently used for PNI repair still cannot achieve the repair effect of autologous nerve transplantation, especially it is difficult to quickly and effectively repair long-distance nerve defects and induce rapid regeneration of new nerve tissue. grow. The fundamental reason is that this type of bridge lacks biological signals that can effectively promote PNI repair. Photopatterning technology provides a powerful means for introducing biochemical signals into biol...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61L27/22A61L27/54A61L27/50
CPCA61L27/227A61L27/50A61L27/54A61L2300/414A61L2430/32C08L89/00
Inventor 凌珏杨宇民顾心逸汤晓璇陈晓莉
Owner NANTONG UNIVERSITY
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