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Aggregation-induced emission nanoparticle and preparation method and application thereof

A technology of aggregation-induced luminescence and nano-particles, which is applied in the field of medical materials, can solve the problems of interfering with the normal physiological functions of cells, the toxicity of quantum dots with heavy metals, and the reduction of fluorescence intensity, so as to achieve cell survival and differentiation ability without interference and good tracer effect, the effect of high luminous efficiency

Active Publication Date: 2016-11-09
SOUTH CHINA UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, fluorescent protein and luciferase require gene transfection, which may interfere with the normal physiological functions of cells; quantum dots are toxic to heavy metals, and their fluorescence intensity decreases sharply during cell differentiation; the fluorescence of traditional organic small molecule dyes has aggregation Induced quenching defect, when stained at high density, its fluorescence will undergo obvious self-quenching

Method used

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  • Aggregation-induced emission nanoparticle and preparation method and application thereof
  • Aggregation-induced emission nanoparticle and preparation method and application thereof
  • Aggregation-induced emission nanoparticle and preparation method and application thereof

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Embodiment 1

[0039] A method for preparing aggregation-induced luminescence nanoparticles, comprising the following steps:

[0040] (1) Distearoylphosphatidylethanolamine-polyethylene glycol-2000 (1mg), distearoylphosphatidylethanolamine-polyethylene glycol-2000-maleic anhydride (1mg), the compound shown in formula I (R 1 -R 9 All hydrogen) (1 mg) was dissolved in 1 mL of tetrahydrofuran, and added to 9 mL of distilled water under ultrasound (80% output, SCIENTZ-II D ultrasonic instrument), and the ultrasound was continued for 2 minutes to prepare AIE nanoparticles with maleic anhydride on the surface.

[0041] (2) Subsequently, tetrahydrofuran was removed by volatilization, and further filtered through a 0.2 μm filter head to remove precipitates and large particles. Cell-penetrating polypeptide Tat(RKKRRQRRRC) (SEQ ID No.1) was added to the filtrate, and after overnight reaction at room temperature, excess polypeptide was removed by dialysis to prepare AIE nanoparticles that could be us...

Embodiment 2

[0044] A method for preparing aggregation-induced luminescence nanoparticles, comprising the following steps:

[0045] (1) distearoylphosphatidylethanolamine-polyethylene glycol-2000 (1mg), distearoylphosphatidylethanolamine-polyethylene glycol-2000-carboxyl (1mg), the compound shown in formula I (R 3 tert-butyl, other substituents are hydrogen) (1 mg) was dissolved in 1 mL of tetrahydrofuran, and added into 9 mL of distilled water under ultrasound (80% output, SCIENTZ-II D ultrasonic instrument), and the surface-modified horses were directly prepared by nano-precipitation method AIE nanoparticles of toric anhydride.

[0046] (2) Subsequently, tetrahydrofuran was removed by volatilization, and further filtered through a 0.2 μm filter head to remove precipitates and large particles. To the filtrate was added the cell penetrating polypeptide Tat(RKKRRQRRRC) (SEQ ID No.1), 69 μg 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride and 78 μg N-hydroxy After reacting with ...

Embodiment 3

[0048] A method for preparing aggregation-induced luminescence nanoparticles, comprising the following steps:

[0049] (1) distearoylphosphatidylethanolamine-polyethylene glycol-2000 (1mg), distearoylphosphatidylethanolamine-polyethylene glycol-2000-amino (1mg), the compound shown in formula I (R 3 Carboxyl, other substituents are hydrogen) (1mg) dissolved in 1mL tetrahydrofuran, under ultrasonic (80% output, SCIENTZ-II D ultrasonic instrument) into 9mL distilled water, directly prepared by nano-precipitation method with maleic anhydride AIE nanoparticles.

[0050] (2) Subsequently, tetrahydrofuran was removed by volatilization, and further filtered through a 0.2 μm filter head to remove precipitates and large particles. To the filtrate was added the cell penetrating polypeptide Tat(RKKRRQRRRC) (SEQ ID No.1), 69 μg 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride and 78 μg N-hydroxy After reacting with sulfosuccinimide overnight at room temperature, the excess pep...

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Abstract

The invention discloses an aggregation-induced emission nanoparticle and preparation method and application thereof. The preparation method comprises the following steps: dissolving an aggregation-induced emission compound shown in a formula I and an amphiphilic polymer in a water-soluble organic solvent, then adding the mixture to water, and conducting ultrasonic reaction; removing organic solvents by volatilization to form nanoparticles loaded with the aggregation-induced emission compound; adding cell penetrating peptides in aqueous solution of the nanoparticle, reacting to form aggregation-induced emission nanoparticles capable of efficiently staining cells. The fluorescent particles can be used for tracing long-term differentiation of stem cells, especially the tracing of mouse bone marrow mesenchymal stem cells in the process of osteogenic differentiation. Compared with the commercial tracer reagent in the prior art, which can only trace 6 generations of cell proliferation, the nanoparticle has a better tracing effect, which can trace to more than 12 generations of cell proliferation. The aggregation-induced emission nanoparticle has the advantages of high cell staining efficiency, long-term tracing ability, high light stability and low cell toxicity.

Description

technical field [0001] The invention belongs to the field of medical materials, and in particular relates to an aggregation-induced luminescent nanoparticle and its preparation method and its application in stem cell tracing, especially its ability in tracing the differentiation process of mouse bone marrow mesenchymal stem cells to osteoblasts use. Background technique [0002] Bone injuries caused by trauma or disease are very common clinically. Traditional clinical treatment is mainly to achieve repair through autologous bone grafting. However, there are many defects in bone grafting, such as limited bone source, long operation time, and comprehensive complications after operation. [0003] In recent years, the application of stem cell therapy in bone repair has attracted increasing attention. For example, bone marrow mesenchymal stem cells are rich in sources, have strong differentiation and proliferation ability, can be cultured, expanded and differentiated in large ...

Claims

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Application Information

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IPC IPC(8): C09K11/07A61K49/00G01N21/64
CPCA61K49/0021C09K11/07G01N21/6486
Inventor 唐本忠高蒙赵祖金任力王迎军陈军建
Owner SOUTH CHINA UNIV OF TECH
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