Preparation method and application of nerve growth factor controlled-release nano-carrier
A nerve growth factor and nanocarrier technology, used in the preparation of drugs for diabetes combined with coronary heart disease, a nerve growth factor sustained release system (nanocarrier) and its preparation, and the field of preparation of medicines for improving the function of diabetic cardiac autonomic and sensory nerves , can solve the problem of unoptimistic treatment effect, and achieve the effect of increasing stability and action time, non-toxic encapsulation and film formation, and good biocompatibility
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Embodiment 1
[0025] Embodiment 1: PLGA-NGF production
[0026] Add 500 μg NGF to 250 μl borate buffer solution (0.05M), weigh 0.5 mg FITC and add it to 250 μl protein system, mix well until FITC is completely dissolved. React for 1 hour at room temperature in the dark, then apply purification column for purification, centrifuge at 1000g for 30-45 seconds, collect the purified protein, and refrigerate at 4°C in the dark. Inner water phase (W 1 ), 200 μl of FITC-labeled NGF was added with 5% PEG; oil phase (O), 25 mg of PLGA was dissolved in 2 ml of a mixed solution of dichloromethane and acetone with a volume ratio of 3:1, and the protein solution was added to the PLGA solution, Prepare water-in-oil emulsion (W 1 / O). Will (W 1 / 0) Emulsion is added in the aqueous solution that contains 0.7%PVA in 10ml, ultrasonic 20min in 55W ice water, obtains water-in-oil-in-water type double emulsion (W 1 / O / W 2 ). Pour the uniform double emulsion into a centrifuge tube, and evaporate in vacuum at...
Embodiment 2
[0027] Example 2: Observation of surface morphology of PLGA-NGF
[0028] Weigh 1 mg of PLGA-NGF nanoparticle freeze-dried powder, add it to 1 ml of distilled water and stir to form a white emulsion, absorb a small amount of emulsion and drop it on the copper grid, then add 2% phosphotungstic acid solution for negative staining, and let it dry naturally. The surface morphology of the particles was then observed under a transmission electron microscope (TEM). see figure 1 .
Embodiment 3
[0029] Embodiment three: the mensuration of nanoparticle average particle size and particle size distribution
[0030] Weigh a little PLGA-NGF nanoparticle freeze-dried powder, dilute it with distilled water to a slightly turbid concentration, and measure the average particle size and For particle size distribution, 14 cycles were taken, three independent samples were taken, and each sample was measured three times.
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