A kind of hierarchical porous functional scaffold material for mobilizing endogenous neural stem cells to repair spinal cord injury and its preparation method and application
A neural stem cell and scaffold material technology, applied in the field of multi-level porous functional scaffold materials and their preparation, to achieve the effects of promoting motor function recovery, promoting regeneration and repair, and controllable strength
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Embodiment 1
[0051] Example 1 Preparation of the multi-level porous functional scaffold material of the present invention (1)
[0052] 1. Chitosan grafted double bond:
[0053] Weigh 2.0 g of chitosan into a three-necked flask, add 150 ml of 2% acetic acid aqueous solution, and dropwise add a small amount of KOH aqueous solution to adjust the pH to 3.8. Then, 3.51 g of glycidyl methacrylate (GMA) was slowly added dropwise, continuously stirred, and reacted at 60° C. for 6 hours. Then pour the reaction mixture into acetone to obtain a large amount of flocculent precipitate, then pour the flocculent into acetonitrile to obtain a powdery precipitate, and finally wash with acetone several times and dissolve it in water, then put it into a dialysis bag with a molecular weight of 3500 Dialyzed for 3 days and freeze-dried to obtain the raw material Chitosan-GMA, which was set aside. The chitosan grafted with double bonds was characterized by infrared spectroscopy and H-NMR.
[0054] result: f...
Embodiment 2
[0066] Example 2 Preparation of the multi-level porous functional scaffold material of the present invention (2)
[0067] 1. Chitosan grafted double bond:
[0068] With embodiment 1.
[0069] 2. Preparation of materials:
[0070] Weigh 50mg Chitosan-GMA into a small glass bottle, then add 25μL BIS (N,N'-methylenebisacrylamide, 10mg / mL) crosslinking agent (0.5% of the monomer), then add 2mg photoinitiator 2-Hydroxy-4'-(2-hydroxyethoxy)-2-methylpropiophenone (4% of the monomer), and finally add 1700 μL H 2 O, stirred at 37°C for 2 hours to dissolve it to form a precursor, that is, a gel-forming precursor.
[0071] After the dissolution is complete, the gel-forming precursor solution is centrifuged at 4500r / min for 4min to remove air bubbles. The solution is then injected into the capillary using an ear wash bulb.
[0072] The capillary injected with the solution was placed in liquid nitrogen at a constant speed (2.5 mm / s) using a lifter for freezing guidance. Immediately a...
Embodiment 3
[0073] Example 3 Preparation of multi-level porous functional scaffold material of the present invention (3)
[0074] 1. Chitosan grafted double bond:
[0075] With embodiment 1.
[0076] 2. Preparation of materials:
[0077] Weigh 50mg Chitosan-GMA into a small glass bottle, then add 50μL BIS (N,N'-methylenebisacrylamide, 10mg / mL) crosslinking agent (1% of the monomer), then add 1mg photoinitiator 2-Hydroxy-4'-(2-hydroxyethoxy)-2-methylpropiophenone (2% of the monomer), and finally add 2200 μL H 2 O, stirred at 37°C for 2 hours to dissolve it to form a precursor, that is, a gel-forming precursor.
[0078] After the dissolution is complete, the gel-forming precursor solution is centrifuged at 4500r / min for 4min to remove air bubbles. The solution is then injected into the capillary using an ear wash bulb.
[0079] Place the capillary injected with the solution in liquid nitrogen at a constant speed (3.5mm / s) for freezing guidance. Immediately after the guidance, the capi...
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