Hollow mesoporous silicon dioxide nanoparticle, hollow mesoporous silicon dioxide nano-carrier and preparation method thereof

A technology of mesoporous silica and nanocarriers, which is applied in the fields of nanocarriers and their preparation, and hollow mesoporous silica nanoparticles, can solve the prospect of limited in vivo application, weak antiserum transfection ability, decreased surface potential, etc. problem, to achieve the effect of vector cytotoxicity, increased transfection efficiency, and complete spherical shape

Active Publication Date: 2017-05-24
GUANGZHOU ZHONGDA NANSHA TECH INNOVATION IND PARK +1
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  • 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

Method used

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  • Hollow mesoporous silicon dioxide nanoparticle, hollow mesoporous silicon dioxide nano-carrier and preparation method thereof
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  • Hollow mesoporous silicon dioxide nanoparticle, hollow mesoporous silicon dioxide nano-carrier and preparation method thereof

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Example Embodiment

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

[0036] (1) Mix alcohols, deionized water and ammonia water at 20-50°C (more preferably 25-35°C) with magnetic stirring;

[0037] (2) continue mixing after ethyl orthosilicate is rapidly added to the product obtained in step 1;

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

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

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

Example Embodiment

[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 this embodiment includes the following steps:

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

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

Example Embodiment

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

[0057] Human colon cancer cells (Lovo) were counted after passage, diluted, and 2*10 cells were added to each well of a 24-well plate. 5 After culturing for 24 hours, the serum-free medium was changed, and the hollow mesoporous silica nanoparticles prepared in Example 1 and the hollow mesoporous silica nanocarriers with a mass ratio of 60:1 to 1.8kDa PEI were precisely weighed. After mixing 120 μg with 2 μg of GFP-DNA prepared in Example 1 for two hours, the obtained hollow mesoporous silica gene nanocarriers (HMSNs-1.8kDaPEI) were cultured with human colon cancer cells (Lovo) for 4 hours, and replaced with serum culture. After culturing for 48 hours, add 200 μL of trypsin to digest the obtained product into a 1.5 mL EP tube, centrifuge at 1200 rpm for 3 min, wash it once with phosphate buffered saline (PBS), and then add 500 μL of PBS and measure it by flow cytometr...

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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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IPC IPC(8): C01B33/18B82Y40/00A61K9/127A61K48/00A61K47/04
CPCA61K47/02A61K48/0008A61K9/1271C01B33/18C01P2004/60
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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