Multifunctional microvesicle, preparation method thereof and application of multifunctional microvesicle
A multifunctional, microbubble technology used in the field of biomedical materials
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[0059] The present invention also provides a method for preparing the multifunctional microbubbles, comprising the following steps:
[0060] (1) dissolving phospholipids and cationic lipids containing photosensitizer functional groups in ethanol to obtain ethanol mixed solution;
[0061] (2) mixing the ethanol mixed solution with physiological saline to obtain a physiological saline system;
[0062] (3) Dialyzing the physiological saline system to obtain a dialysate;
[0063] (4) mixing the dialysate and a stabilizer to obtain a stable system;
[0064] (5) Add a contrast agent to the stable system, and obtain primary microbubbles after shaking;
[0065] (6) Mixing the primary microbubbles and nucleic acid to obtain multifunctional microbubbles.
[0066] The invention dissolves phospholipid and cationic lipid containing photosensitizer functional group in ethanol to obtain ethanol mixed solution. In the present invention, the molar ratio of phospholipids to cationic lipids ...
Embodiment 1
[0079] Distearoylphosphatidylcholine (DSPC), cholesterol (cholesterol), distearoylphosphatidylethanolamine-polyethylene glycol 2000 (DSPE-PEG2000) and cationic tetraphenyl porphyrin lipid (CPGL) were Molar ratio (35%: 10%: 5%: 50%) is mixed and dissolved in ethanol, wherein, the concentration of the cationic porphyrin lipid in ethanol is 1 mg / mL; then adopt ethanol injection method, at 50 ℃ In a water bath at an ultrasonic frequency of 100MHz, inject 0.15mL of the above mixture into 0.8ml of normal saline, and treat with ultrasound for 20 minutes; place the solution obtained above in a dialysis bag with a molecular weight cut-off of 8000Da, dialyze for 3h, and add glycerol after taking it out Mix with 100 μL of propylene glycol, put the mixture into a 3.5mL vial, fill with perfluoropropane (C 3 f 8 ) gas to exhaust the air, and to normal pressure, shake the oscillator for 45s, add an appropriate amount of siRNAHIF1α according to N / P=15:1, let it stand at room temperature for ...
Embodiment 2
[0082]Distearoyl phosphatidylcholine (DSPC), distearoyl phosphatidylethanolamine-polyethylene glycol 2000 (DSPE-PEG2000), cationic tetraphenyl porphyrin lipid (CPGL) according to a certain molar ratio (45% :5%:50%) mixed and dissolved in ethanol, wherein, the concentration of the cationic porphyrin lipid in ethanol is 1mg / mL; 0.2 mL of the above mixture was injected into 0.8 mL of water, and ultrasonically treated for 20 minutes; the solution obtained above was placed in a dialysis bag with a molecular weight cut-off of 14,000 Da, dialyzed for 2 hours, and then 100 μL of glycerin and propylene glycol were added and mixed well. Put the mixed solution into a 3.5mL vial, fill it with perfluorobutane gas to drive out the air, and bring it to normal pressure, shake the oscillator for 45s, add an appropriate amount of siRNAHIF1α according to N / P=30:1, and let it stand at room temperature for 15min. After separation and purification, multifunctional microbubbles (siHIF@CpMBs) integra...
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