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Fibrous protein targeted multi-modal nano particles for micro-thrombus detection and application thereof

A technology of fibrin and nanoparticles, applied in the field of clinical diagnosis and molecular imaging, to avoid oxidative inactivation, ensure specificity and sensitivity, and ensure spatial resolution and sensitivity

Inactive Publication Date: 2014-09-03
ZHONGSHAN HOSPITAL FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] However, there is no connection or combination of superparamagnetic iron ferric oxide nanoparticles, near-infrared fluorescent material IR783, and tumor-homing short peptide CREKA to prepare multimodal Related reports on state nanoprobes

Method used

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  • Fibrous protein targeted multi-modal nano particles for micro-thrombus detection and application thereof
  • Fibrous protein targeted multi-modal nano particles for micro-thrombus detection and application thereof
  • Fibrous protein targeted multi-modal nano particles for micro-thrombus detection and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0063] Example 1 Synthesis of multifunctional nanoprobe IR783-R-SPIO-PEG-CREKA (1)

[0064] The schematic diagram of the synthesis process is attached figure 1 . The fluorescent dye is carried on the surface of SPIO, and then SPIO is modified with PEG, and Mal-PEG is covalently combined with CREKA to obtain IR783-R-SPIO-PEG-CREKA (SPIO-CREKA for short). Specific steps are as follows:

[0065] (1) Synthesis of IR783-SPIO: Take 20mg SPIO, that is, 4ml SPIO solution (5mg / ml) and replace it with PBS buffer at pH 7.4 after ultrafiltration; take 20μl IR783-NHS (10mg / ml) and add it to the SPIO solution, 25°C, 50W sonication for 2h; the reaction resultant was passed through a Hitrap column equilibrated with PBS buffer at pH 7.4 to remove unlinked IR783; and the IR783-NHS linking efficiency was measured. See attached figure 2 a.

[0066] (2) Synthesis of IR783-R-SPIO: Add 40 μl rhodamine-NHS to the IR783-SPIO solution, 25°C, 50W ultrasonication for 2 hours; take the reaction ...

Embodiment 2

[0069] Example 2 Particle size distribution and Zeta potential of multifunctional nanoprobes

[0070] Take 800 μl of SPIO, IR783-R-SPIO, IR783-R-SPIO-PEG and IR783-R-SPIO-PEG-CREKA solutions respectively, and use a particle size / Zeta potential measuring instrument to measure the light scattering particle size and particle distribution width. Take 1ml of SPIO, IR783-R-SPIO, IR783-R-SPIO-PEG, and IR783-R-SPIO-PEG-CREKA solutions, respectively, and replace the buffer with a Hitrap column equilibrated with 0.001M NaCl solution in advance. Diameter / Zeta potential measuring instrument to measure its Zeta potential.

[0071] The results show that the hydrodynamic particle size of the nanoprobe measured by dynamic light scattering method is 103.6±4.7nm, and the average Zeta potential is -2.2±0.7mV. See attached image 3 .

Embodiment 3

[0072] Example 3 Detection of the binding ability of multifunctional nanoprobes to thrombus in vitro

[0073]Fluorescence microscope, optical imaging system and MRI imaging system were used to detect the in vitro binding ability of multifunctional nanoprobes to thrombus fibrin. Specific steps are as follows:

[0074] (1) Fluorescence microscope detection: add 20 μl human fresh plasma, 2 μl CaCl on the glass slide 2 (0.4mol / L) solution and 2μl thrombin (0.1U / μl); place the slide in a 37°C incubator and incubate for 60 minutes; add 20μl PBS, SPIO-PEG (5mg / ml SPIO) and SPIO-CREKA respectively After (5mg / ml SPIO), place in a 37°C incubator and incubate for 15 minutes; wash with PBS for 5min×3 times and observe under a fluorescence microscope (N3 channel, excitation 546±6nm, excitation 600±20nm).

[0075] The results showed that in the SPIO-CREKA group, red fluorescence could be observed on the surface of the fibrin plug, while there was no obvious red fluorescence in the PBS g...

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Abstract

The invention relates to fibrous protein targeted multi-modal nano particles for micro-thrombus detection and application thereof. The multi-modal nano particles comprise a magnetic resonance probe, a near-infrared fluorescent probe, fluorescent dye, a surface modification molecule and a fibrous protein targeted molecule, wherein the magnetic resonance probe refers to superparamagnetic ferroferric oxide nano particles; the near-infrared fluorescent probe is IR783; the fluorescent dye is rhodamine; the surface modification molecule is polyethylene glycol; and the fibrous protein targeted molecule is homing peptide. According to the multi-modal nano particles, the magnetic resonance probe and the near-infrared fluorescent probe are combined with each other, multi-modal synchronous noninvasive detection of thrombus can be realized, the detected space resolution and sensitivity are guaranteed, the high-density CREKA is distributed on the nano particles, and the specificity and sensitivity are further guaranteed. Therefore, the nano particles are particularly suitable for micro-thrombus detection with small size and disperse distribution, the fluorescent dye is connected to the nano particles, so that in-vitro detection is conveniently realized.

Description

technical field [0001] The invention relates to the technical field of clinical diagnosis and molecular imaging, in particular to the synthesis of a multifunctional nanoparticle with active targeting effect on fibrin, which is used for non-invasive multimodal detection of microthrombosis. Background technique [0002] Thrombosis is the main pathogenesis of acute ischemic cardiovascular and cerebrovascular diseases (such as unstable angina, acute myocardial infarction, stroke and pulmonary embolism, etc.). Recent studies have shown that microthrombosis also plays an important role in cardiovascular diseases. Participate in the occurrence of coronary no-reflow, cardiac X syndrome, coronary microvascular spasm and other diseases. In addition, one month before the onset of acute myocardial infarction, microthrombi had appeared on unstable atherosclerotic plaques. [0003] Commonly used clinical imaging methods (such as ultrasound, CTA, intravascular ultrasound (IVUS), optical ...

Claims

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Application Information

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IPC IPC(8): A61K49/12A61K49/00A61K49/18A61K9/14
Inventor 葛均波钱菊英黄浙勇宋亚楠庞志清
Owner ZHONGSHAN HOSPITAL FUDAN UNIV
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