Method for preparing mesoporous silica nanoparticles loaded with calcein and wrapped by cationic liposome

A technology of mesoporous silica and cationic liposomes, applied in the fields of biotechnology and medicine, can solve the problems of drug leakage, reduce drug efficacy, and take a long time, and achieve high-efficiency live cell labeling, high live cell labeling efficiency, low cytotoxic effect

Inactive Publication Date: 2018-09-21
TIANJIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the toxicity of silicon dioxide itself, it will cause certain damage to the cells after being added to the cells.
The absorption of silica nanoparticles by cells usually goes through the proce

Method used

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  • Method for preparing mesoporous silica nanoparticles loaded with calcein and wrapped by cationic liposome
  • Method for preparing mesoporous silica nanoparticles loaded with calcein and wrapped by cationic liposome
  • Method for preparing mesoporous silica nanoparticles loaded with calcein and wrapped by cationic liposome

Examples

Experimental program
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Effect test

Embodiment 1

[0035] (1) Preparation of mesoporous silica nanoparticles loaded with calcein: use deionized water to configure calcein into a solution with a concentration of 250 micromolar, add mesoporous silica nanoparticles with a mass ratio of 1:5, and mix well , magnetically stirred at 400 rpm in the dark for 12 hours, centrifuged at 15,000 rpm for 15 minutes, carefully discarded the supernatant, and resuspended the precipitate in double distilled water to 5 mg / ml to obtain calcein-loaded mesoporous silica nanoparticles. particles.

[0036] (2) Preparation of cationic liposomes with uniform particle size: Weigh (2,3-dioleoyl-propyl)-trimethylamine and add chloroform to prepare a solution with a concentration of 2 mg / ml. Set the temperature of the water bath of the rotary evaporator to 36° C., turn on the vacuum pump to vacuum for 30 minutes, and a layer of milky white covering is formed at the bottom of the round bottom flask, which is a monolayer cationic liposome. Add 50% mass fracti...

Embodiment 2

[0040](1) Preparation of mesoporous silica nanoparticles loaded with calcein: use deionized water to configure calcein into a solution with a concentration of 260 micromolar, add mesoporous silica nanoparticles with a mass ratio of 1:8, and mix well , protected from light at 350 rpm magnetic stirring for 10 hours, centrifuged at 12,000 rpm for 13 minutes, carefully discarded the supernatant, and resuspended the precipitate with double distilled water to 3 mg / ml to obtain calcein-loaded mesoporous silica nanoparticles. particles.

[0041] (2) Preparation of cationic liposomes with uniform particle size: Weigh (2,3-dioleoyl-propyl)-trimethylamine and add chloroform to prepare a solution with a concentration of 8 mg / ml. Set the temperature of the water bath of the rotary evaporator to 40°C, turn on the vacuum pump to vacuum for 35 minutes, and a layer of milky white covering is formed at the bottom of the round bottom flask, which is a monolayer cationic liposome. Add 50% mass f...

Embodiment 3

[0045] (1) Preparation of mesoporous silica nanoparticles loaded with calcein: use deionized water to configure calcein into a solution with a concentration of 300 micromolar, add mesoporous silica nanoparticles with a mass ratio of 1:10, and mix well , magnetically stirred at 300 rpm in the dark for 6 hours, centrifuged at 10,000 rpm for 10 minutes, carefully discarded the supernatant, and resuspended the precipitate in double-distilled water to 1 mg / ml to obtain calcein-loaded mesoporous silica nanoparticles. particles.

[0046] (2) Preparation of cationic liposomes with uniform particle size: Weigh (2,3-dioleoyl-propyl)-trimethylamine and add chloroform to prepare a solution with a concentration of 10 mg / ml. Set the temperature of the water bath of the rotary evaporator to 42° C., turn on the vacuum pump to vacuum for 40 minutes, and a layer of milky white covering is formed at the bottom of the round bottom flask, which is a monolayer cationic liposome. Add 50% mass fract...

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Abstract

The invention relates to a method for preparing mesoporous silica nanoparticles loaded with calcein and wrapped by cationic liposome. The Calcein is loaded into mesoporous silica; the cationic liposome with uniform particle size is prepared to encapsulate the outside of the mesoporous silica nanoparticles loaded with the calcein. The cationic liposome prepared by the method encapsulates the calcein-loaded mesoporous silica nanoparticles, the prepared mesoporous silica nanoparticles loaded with calcein and wrapped by the cationic liposome have particle diameters of 80-130 microns, after cell addition, the cell survival rate is 80%-90%, the efficiency of viable cell labeling observed by an inverted fluorescence microscopy is about 90%-95%, the positions of viable cells can be judged by observing the distribution of the calcein in the nanoparticles, and high-efficient viable cell labeling is achieved.

Description

technical field [0001] The invention relates to a preparation method of nanoparticles capable of marking and positioning live cells, belonging to the technical fields of biotechnology and medicine. In particular, it relates to a preparation method for encapsulating calcein-loaded mesoporous silicon dioxide nanoparticles with cationic liposomes. Background technique [0002] Calcein is usually used in fields such as complexation indicators, fluorescent indicators, etc. Due to its hydrophilicity, it does not easily penetrate the cell membrane, and stays in the intercellular space after adding cells, so it cannot be used for live cell labeling. The current method of using calcein to label living cells is to strengthen the hydrophobic modification on the surface of calcein to form the living cell dye Calcein (calcein)-AM, which will be hydrolyzed by the esterase in the living cells after the hydrophobic action penetrates the living cell membrane. The calcein stays in the cells ...

Claims

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Application Information

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IPC IPC(8): G01N33/532
CPCG01N33/532
Inventor 常津潘惠卓王汉杰谌红彬王天歌张超楠王镐锋郝亚锋
Owner TIANJIN UNIV
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