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Targeting magnetic fluorescent nano-vector carrying Notch-1 shRNA as well as preparation method and application of nano-vector

A technology of fluorescent nanometer and nanocarrier, which is applied in preparations for in vivo experiments, medical preparations with non-active ingredients, and pharmaceutical formulations, etc. It can solve the problems of loss of magnetism and low colloidal stability, and achieve low cost and long storage time. long-lasting and apoptosis-promoting effects

Inactive Publication Date: 2015-01-21
UNIV OF ELECTRONICS SCI & TECH OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, in the application of biomedicine, there are many unavoidable shortcomings of ferric oxide nanoparticles. For example, after the particles enter body fluids and cells, they are easy to aggregate in the cells to form larger particle clusters. Compared with other particles, nanoparticles have low colloidal stability, and are prone to oxidation in the intracellular environment, causing them to lose their original magnetic properties

Method used

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  • Targeting magnetic fluorescent nano-vector carrying Notch-1 shRNA as well as preparation method and application of nano-vector
  • Targeting magnetic fluorescent nano-vector carrying Notch-1 shRNA as well as preparation method and application of nano-vector
  • Targeting magnetic fluorescent nano-vector carrying Notch-1 shRNA as well as preparation method and application of nano-vector

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1: Synthesis of targeted magnetic fluorescent nanocarriers

[0032] ①The nanoparticles are Fe 3 o 4 as the core, with SiO 2 As the shell, and doped with fluorescent dye FITC, the surface of the carrier is modified with PEI-FA to adsorb Notch-1 shRNA gene. The schematic diagram of its synthesis is as figure 1 shown.

[0033] ②Nanoparticles have uniform particle size, regular shape, uniform spherical shape, particle size of about 60-70nm, and good dispersion in aqueous solution.

[0034] ③The nanoparticles not only have the functions of nuclear magnetic resonance imaging and fluorescence imaging. The surface-modified high-molecular polymer PEI-FA with targeting function can target human breast cancer cells (MDA-MB-231) with high folate receptor expression, release the carried gene, and silence the corresponding protein. Express.

Embodiment 2

[0035] Example 2: Fe 3 o 4 SiO 2 Preparation of (FITC) nanoparticles

[0036] ① Place 1ml of double distilled water, 1-5ml of magnetic fluid, 5-10ml of absolute ethanol and 200μl of ammonia water in a shaker at 300rpm for 10-30min.

[0037] ② Quickly add 5-10 μl tetraethyl orthosilicate (TEOS) to the above reaction system, keep away from light, place in a shaker, and shake at 300 rpm for 24 hours.

[0038] ③Add 1-5ml FITC-APTMS solution to the above reaction system, place it in a shaker, and continue shaking at 300rpm for 48h.

[0039] ④ Distribute the above solution evenly into EP tubes, centrifuge at 12000rpm for 5min, discard the supernatant, add 1ml of double distilled water, resuspend by ultrasonication, and wash with double distilled water for 5 times. Fe 3 o 4 SiO 2 (FITC) solid was dissolved in double distilled water and stored at 4°C until use.

[0040] The nanoparticles obtained in this example are uniform in size, uniform in spherical shape, good in dispersi...

Embodiment 3

[0041] Example 3: Fe 3 o 4 SiO 2 Preparation of (FITC) / PEI-FA / Notch-1 shRNA

[0042] ① Weigh 20 mg of folic acid powder, add 5 ml of DMSO to completely dissolve the folic acid, and the concentration is 4 mg / ml. Take 5-10ml of folic acid solution, add 4-10ml of DCC and 4-10ml of NHS respectively, place in a shaker at 300rpm, and shake for 10min.

[0043] ② Add an appropriate amount of PEI (20mg / ml) and continue shaking for 12 hours. After the reaction is completed, place the reactant in a dialysis bag and dialyze for 48 hours. And the liquid in the dialysis bag was placed in a glass bottle and freeze-dried.

[0044] ③Take 2 g of the dried solid and dissolve it in double distilled water to obtain a 20 mg / ml PEI-FA suspension

[0045] 4. first prepare Fe by the method of embodiment 2 3 o 4 SiO 2 (FITC).

[0046] ⑤ The above PEI-FA and Fe 3 o 4 SiO 2 (FITC) was mixed according to the mass ratio of 1:5-10, and the magnetic stirring speed was 300rpm for 2h. After the re...

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Abstract

The invention relates to a preparation method and biomedical application of a Fe3O4SiO2 (FITC) nano-vector integrated with magnetic resonance imaging, fluorescence imaging and targeting gene therapy. The nano-vector is prepared by virtue of the following steps: taking silicone having low toxicity and excellent biocompatibility as a raw material; firstly, synthesizing Fe3O4SiO2 nanoparticles doped with a fluorescent dye by use of an improved St.ber method, and linking polyethyleneimine (PEI) capable of improving the transfection efficiency with folic acid (FA) capable of targeting a folate receptor by virtue of an amidation reaction to form a PEI-FA polymer; further modifying the Fe3O4SiO2 (FITC) nanoparticles by use of the PEI-FA polymer, and adsorbing the shRNA of Notch-1 to form the compound nano-vector. The nano-vector has the functions of magnetic resonance imaging and fluorescence imaging, and further is capable of conveying a targeting gene to a tumor part to silence the expression of a target gene and inhibit the growth of the tumor.

Description

technical field [0001] The invention belongs to the field of biological materials and relates to the synthesis of a nuclear magnetic resonance contrast agent, in particular to a method for preparing a targeted magnetic fluorescent nanocarrier carrying Notch-1 shRNA, and its application in nuclear magnetic resonance imaging, fluorescent imaging and targeted gene therapy, etc. applications in biomedicine. Background technique [0002] Magnetic resonance imaging is currently a widely used imaging tool in biomedicine, with the advantages of high spatial resolution, low biological invasiveness, and no ionizing radiation. The use of nuclear magnetic resonance contrast agents can significantly improve the specificity and sensitivity of imaging, thereby improving the imaging effect of MRI. 2 Contrast agents for weighted imaging. Ferroferric oxide nanoparticles are a kind of magnetic nanoparticles, which have multiple uses such as drug delivery, cell separation, and nuclear magneti...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K49/12A61K49/00A61K48/00A61K47/34A61P35/00
Inventor 刘贻尧李亭亭李颖杨红
Owner UNIV OF ELECTRONICS SCI & TECH OF CHINA
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