A preparation method of fluorescent silica nanoparticles across the blood-brain barrier membrane with different particle sizes
A blood-brain barrier and nanoparticle technology, applied in the field of nano-biomedical materials, can solve the problems of influence of treatment effect, low ability of drug delivery carrier to cross the BBB, low targeting ability, etc., achieve good physical and chemical stability, meet production requirements and application, high biosafety effect
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Embodiment 1
[0028] 1. Preparation of Fluorescent APTES Precursors
[0029] Take 2.0 mg of 6-carboxyfluorescein and disperse it in 1.0 mL of ethanol, sonicate for a short period of time. After the solution is clarified, add 80 microliters of APTES under the action of a magnetic stirrer and react in the dark for 24 hours to prepare fluorescent APTES. precursor.
[0030] 2. Preparation of FSiNPs
[0031] Dissolve 0.5 mL TEOS and 0.1 mL fluorescent APTES precursor in a mixed solvent of ethanol / water (ethanol: water = 0.5:1) in sequence, then mix with 0.1 mL 25% concentrated ammonia water, and stir for 24 hours. Then, they were washed with ethanol and water three times in sequence, and finally stored in DMF at 4 °C to prepare FSiNPs.
[0032] 3. FSiNPs-PEG 2000 - Preparation of Mal
[0033] Add 1.6 μl of triethylamine to the above DMF solution of FSiNPs, stir well, add 20 mg of NHS-PEG 2000 -Mal, react at room temperature for 2 hours. Wash with DMF and water by centrifugation, and store ...
Embodiment 2
[0038] 1. Preparation of Fluorescent APTES Precursors
[0039] Take 2.0 mg of fluorescein cy3, disperse it in 1.0 mL of ethanol, and sonicate for a short period of time. After the solution is clarified, add 80 microliters of APTES under the action of a magnetic stirrer, and react in the dark for 24 hours to prepare the fluorescent APTES precursor .
[0040] 2. Preparation of FSiNPs
[0041] Dissolve 0.5 mL TEOS and 0.1 mL fluorescent APTES precursor in a mixed solvent of ethanol / water (ethanol: water = 1:1), then mix with 0.2 mL 25% concentrated ammonia water, and stir for 24 hours. Then, they were washed with ethanol and water three times in sequence, and finally stored in DMF at 4 °C to prepare FSiNPs.
[0042] 3. FSiNPs-PEG 2000 - Preparation of Mal
[0043] Add 1.6 μl of triethylamine to the above DMF solution of FSiNPs, stir well, add 20 mg of NHS-PEG 2000-Mal, react at room temperature for 2 hours. Wash with DMF and water by centrifugation, and store in 1.0 mL of 1...
Embodiment 3
[0048] 1. Preparation of Fluorescent APTES Precursors
[0049] Take 2.0 mg of fluorescein cy5.5, disperse it in 1.0 mL of ethanol, sonicate for a short period of time, after the solution is clarified, add 80 microliters of APTES under the action of a magnetic stirrer, and react in the dark for 24 hours to prepare fluorescent APTES precursor.
[0050] 2. Preparation of FSiNPs
[0051] Dissolve 0.5 mL TEOS and 0.1 mL fluorescent APTES precursor in a mixed solvent of ethanol / water (ethanol:water=1.5:1) in sequence, then mix with 0.5 mL 25% concentrated ammonia water, and stir for 24 hours. Then, they were washed with ethanol and water three times in sequence, and finally stored in DMF at 4 °C to prepare FSiNPs.
[0052] 3. FSiNPs-PEG 2000 - Preparation of Mal
[0053] Add 1.6 μl of triethylamine to the above DMF solution of FSiNPs, stir well, add 20 mg of NHS-PEG 2000 -Mal, react at room temperature for 2 hours. Wash with DMF and water by centrifugation, and store in 1.0 mL...
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