Alanine modified fullerene nano-material, preparation method thereof and application of alanine modified fullerene nano-material in field of neural restoration
A nanomaterial and nerve repair technology, applied in the direction of fullerene, nanotechnology for materials and surface science, nanocarbon, etc., to achieve the effect of treating central nervous system diseases, promoting proliferation, and promoting the proliferation of neural stem cells
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preparation example Construction
[0051] Preferably, the specific steps of the preparation method are as follows: configure the alkaline alcohol solution of alanine, add the alkaline alcohol solution of alanine dropwise into the fullerene solution under stirring, and then add four Butyl ammonium hydroxide is used to obtain a reaction system; the reaction system is placed at 20-30° C. in a dark place until the reaction is complete.
[0052] Further, the method for judging the completion of the reaction includes that the upper layer of the reaction system turns purple into brownish yellow, and the lower layer turns dark brown.
[0053] Further, the preparation method also includes a separation and purification step: obtain the upper organic layer, adjust the pH to neutral, put it into a dialysis bag with a molecular weight cut-off of 90-110 Daltons, and perform dialysis against water. After the dialysis is completed, filter to remove possible insoluble matter, retain the filtrate, and freeze-dry it or dry it in ...
Embodiment 1
[0068] Preparation and purification of fullerenes modified by alanine in embodiment 1
[0069] Preparation: configure into C 60 The ortho-xylene solution (5mg / ml) is standby (5mg / ml), gets a 100ml beaker, takes by weighing 0.884 gram alanine and joins in the NaOH of 10ml, is mixed with the solution of 1g / ml, then adds 30ml ethanol, obtains solution A. Measure 60ml of C 60 The o-xylene solution was placed in a round-bottomed flask, and solution A was added dropwise to C while stirring. 60 To the solution, 10 drops of tetrabutylammonium hydroxide (TBAH) with a mass fraction of 40% were added to obtain solution B. Place solution B in a dark place at 25°C for reaction until the reaction is complete (the upper layer of purple basically disappears and turns brownish yellow, and the lower layer is dark brown.)
[0070] Separation and purification: separate and remove the upper organic layer, adjust the pH to 7.0, and perform dialysis. After the dialysis, the precipitate was remov...
Embodiment 2
[0072] Example 2 Neural stem cells promote activity
[0073] 1. Extraction of primary neural stem cells
[0074] The 13.5-day-pregnant Wistar rats were killed by decapitation, and put into a tray filled with alcohol for disinfection to ensure a sterile environment. Cut open the abdomen of the rat with scissors, find the beaded embryo, and put it into a 6cm dish filled with DMEM washing solution. The placenta was peeled off, and the fetal mice were separated, and the whole operation was performed on ice. Transfer the fetal mouse to a new 6cm dish, and carefully separate the whole brain of the fetal mouse with micro forceps under a microscope. Then the meninges and red blood vessels were peeled off, the brain tissue was crushed as much as possible with tweezers, and DMEM was poured on the tissue with a straw, sucked back immediately, and placed in a 15mL centrifuge tube. After all fetal mouse brain tissue was collected, centrifuge at 800 rpm for 3 minutes, and carefully disca...
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