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Hollow mesoporous silica nanoparticles, nanocarriers and preparation methods thereof

A mesoporous silica and nano-carrier technology, applied in the direction of silica, silicon oxide, nanotechnology, etc., can solve the problems of limiting the prospect of in vivo application, weak transfection ability of antiserum, and decreased transfection efficiency, achieving Under the cytotoxicity of the vector, the effect of increasing the transfection efficiency and high gene transfection efficiency

Active Publication Date: 2020-06-16
GUANGZHOU ZHONGDA NANSHA TECH INNOVATION IND PARK +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among them, liposome is a relatively mature in vitro transfection carrier, but its stability is poor; cationic polymer carrier usually has a good in vitro gene transfection effect, but this type of material has obvious toxicity, and its transfection ability under serum conditions Poor, which limits the prospect of its in vivo application; Inorganic nanomaterials have good stability and are easy to modify, etc., and can be used as gene delivery carriers. The disadvantage is that the transfection efficiency is usually low
[0003] Among inorganic nanocarriers, mesoporous silica nanoparticles (MSNs) have unique advantages such as low cytotoxicity, good stability, easy modification and large specific surface area, pore volume and ordered pore structure. It has received extensive attention in the field of drug and gene delivery. The disadvantage is that the surface potential of nanoparticles loaded with genes will decrease, which is not conducive to the uptake of cells, and after the system enters cells, lysosomes will interfere with its transfection, causing transfection Efficiency drops
On the other hand, polyethyleneimine (PEI), as a commonly used cationic polymer carrier, can cause the "proton sponge effect", which can avoid the damage of lysosomes to genes, thereby improving the transfection efficiency, but its toxicity Defects such as the weak transfection ability of Yamato antiserum cannot be ignored

Method used

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  • Hollow mesoporous silica nanoparticles, nanocarriers and preparation methods thereof
  • Hollow mesoporous silica nanoparticles, nanocarriers and preparation methods thereof
  • Hollow mesoporous silica nanoparticles, nanocarriers and preparation methods thereof

Examples

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preparation example Construction

[0035] In one of the embodiments, the present invention provides a method for preparing hollow mesoporous silica nanoparticles, comprising the following steps:

[0036] (1) Alcohols, deionized water, and ammonia water are magnetically stirred and mixed at 20-50° C. (more preferably 25-35° C.);

[0037] (2) Continue mixing after adding tetraethyl orthosilicate rapidly to the product obtained in step 1;

[0038] (3) Continue mixing after adding premixed tetraethyl orthosilicate and octadecyltrimethoxysilane to the product obtained in step 2;

[0039] (4) After the obtained product is centrifuged, the lower layer of the precipitate is etched with sodium carbonate at 20-100° C. (more preferably 70-90° C.);

[0040] (5) Vacuum-dry the obtained product and calcinate at 300-600°C (more preferably 500-600°C) to obtain the hollow mesoporous silica nanoparticles; the mass ratio of ethanol, deionized water, and ammonia water For: 65-75:10:2-4. In step (2), the volume ratio of the cons...

Embodiment 1

[0043] Example 1: Preparation of Green Fluorescent Protein Hollow Mesoporous Silica Nanocarriers

[0044] The preparation method of the green fluorescent protein hollow mesoporous silica nanocarrier of the present embodiment comprises the following steps:

[0045] 1. Preparation of green fluorescent protein DNA (GFP-DNA)

[0046]Take 2.5g LB medium powder, add 100mL distilled water, and steam sterilize under 15psi high pressure for 20min. The 100 mL LB medium contains 1 g of peptone, 0.5 g of yeast, and 1 g of sodium chloride. Add kanamycin (to make the final concentration 50 μg / mL) and 1 mL of Escherichia coli transformed with green fluorescent protein gene (pEGFP) into 100 mL of sterilized LB medium in a biological safety cabinet. Incubate at 37.0°C with shaking at 200rpm for 14-16h. Add the bacterial solution cultured overnight into a centrifuge tube, centrifuge at room temperature at 4000 rpm for 6 minutes to collect the bacterial cells, discard the supernatant, and dry...

Embodiment 2

[0056] Example 2: Transfection efficiency of hollow mesoporous silica nanocarriers measured by flow cytometry

[0057] Count the human colon cancer cells (Lovo) after passage, dilute, add 2*10 per well of 24-well plate 5 After culturing for 24 hours, change the serum-free medium, and accurately weigh the hollow mesoporous silica nanoparticle prepared in Example 1 and the hollow mesoporous silica nanocarrier with a mass ratio of 1.8kDa PEI of 60:1 After 120 μg was mixed with 2 μg of GFP-DNA prepared in Example 1 for two hours, the obtained hollow mesoporous silica gene nanocarrier (HMSNs-1.8kDaPEI) was cultured with human colon cancer cells (Lovo) for 4 hours, and replaced with serum culture After 48 hours of culture, 200 μL of trypsin was added to digest the product. The resulting product was placed in a 1.5 mL EP tube, centrifuged at 1200 rpm for 3 minutes, washed once with phosphate buffered saline (PBS), and then 500 μL of PBS was added and measured by flow cytometry. The ...

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Abstract

The invention relates to a hollow mesoporous silicon dioxide nanoparticle and a preparation method thereof. The preparation method comprises the following steps: magnetically stirring and mixing ethanol, deionized water and ammonia water at a certain temperature, adding ethyl orthosilicate for continuous reaction, adding premixed ethyl orthosilicate and trimethoxyoctadecylsilane for continuous reaction, vacuum drying an obtained mixture after being etched by sodium carbonate, calcining at 550 DEG C to obtain the hollow mesoporous silicon dioxide nanoparticle. The nanoparticle is dispersed in ultrapure water, and polymine is added to obtain a hollow mesoporous silicon dioxide nano-carrier after mixing; and the hollow mesoporous silicon dioxide nano-carrier is mixed with a gene to obtain a hollow mesoporous silicon dioxide gene nano-carrier. The hollow mesoporous silicon dioxide nano-carrier prepared by the preparation method disclosed by the invention has the advantages of good dispersity, high gene loading capacity and high transfection efficiency (twice of 25kDa PEI), and has a practical value of clinical application.

Description

technical field [0001] The invention relates to the field of pharmaceutical preparations, in particular to a hollow mesoporous silicon dioxide nanoparticle, a nanocarrier and a preparation method thereof. Background technique [0002] Gene therapy refers to the transfer of exogenous genes with therapeutic effects into specific cells in a certain way to achieve the purpose of treatment. Among them, how to effectively deliver the target gene to cells and exert curative effect is the key to this therapy. Gene delivery usually requires the help of vectors, and the current gene vectors are often divided into two types: viral vectors and non-viral vectors. Although viral vectors can achieve high transfection efficiency, high immunogenicity, possible carcinogenicity and other safety issues limit the wide application of such vectors. Non-viral vectors have attracted the attention of researchers due to their low immunogenicity, high safety, and convenient preparation. This type of ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C01B33/18B82Y40/00A61K9/127A61K48/00A61K47/04
CPCA61K9/1271A61K47/02A61K48/0008C01B33/18C01P2004/60
Inventor 吴传斌湛正文权桂兰潘昕陈航平
Owner GUANGZHOU ZHONGDA NANSHA TECH INNOVATION IND PARK
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