A method for tracing deep cervical lymph nodes and meningeal lymphatic vessels using nanoprobes

A nano-fluorescent probe and fluorescence technology, applied in the field of nano-probes, can solve problems such as presenting the complete structure of the central nervous system

Active Publication Date: 2021-06-08
SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This major discovery fills the gap in the central nervous system, but there is still no definite evidence to present the complete structure of the central nervous system

Method used

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  • A method for tracing deep cervical lymph nodes and meningeal lymphatic vessels using nanoprobes
  • A method for tracing deep cervical lymph nodes and meningeal lymphatic vessels using nanoprobes
  • A method for tracing deep cervical lymph nodes and meningeal lymphatic vessels using nanoprobes

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1 Preparation and characterization of nanoprobes loaded with Cy5

[0027] In this example, by optimizing the ratio of aqueous phase / organic phase and the liquid flow rate of each channel, a nanoprobe with a particle size of 42.4 nm and a surface potential of -21.4 mV was finally prepared. The specific implementation steps are as follows: ① Take 6 mg PLGA, 0.9mg Cy5 was dissolved in 3 mL organic solvent acetonitrile, vortexed and mixed; 15 mg PF127 and 1 mg DSPE-PEG were dissolved in 3 mL ultrapure water, shaken and mixed; ②The acetonitrile solution containing PLGA and ICG was passed through Channel 1 (flow rate 10 mL / min), aqueous solution containing PF127 and DSPE-PEG passed through channel 2 (flow rate 10 mL / min), 12 mL ultrapure water passed through channels 3 and 4 (flow rate 40 mL / min), respectively, and sent Perform high-speed mixing and solvent exchange in a multi-channel vortex mixer to obtain unpurified nanoparticles; ③Use a dialysis bag with a molecula...

Embodiment 2

[0028] Example 2 Real-time imaging of deep cervical lymph nodes and meningeal lymph nodes after injection of nanoprobe CNP

[0029] In this example, the nanoprobe prepared in Example 1 (the concentration of Cy5 is 200 μg / mL) was injected subcutaneously in the right neck of BALB / c mice, and the deep layer of the neck of the mice was surgically exposed Lymph nodes and meningeal lymphatic vessels were observed in real time using an in vivo confocal microscope for changes in their structure and drug concentration. The collective implementation steps are as follows: ① The mice were anesthetized with isoflurane, and 0.2 mL of the nanoprobe CNP was subcutaneously injected near the neck bone of the mouse on the right side of the neck, so that the dose of the probe received by the mice was 2 mg / kg; ② The mice were cut open. Neck skin, blunt dissection of mouse neck musculature, exposing neck deep lymph nodes and nearby meningeal lymphatic vessels; Real-time imaging of deep lymph nodes...

Embodiment 3

[0030] Example 3 Dosage of Nanoprobe CNP in Peripheral Lymph Nodes at Different Time Points

[0031] This example aims to explore the time point when the nanoprobe reaches the highest dose enrichment in the deep cervical lymph nodes. The collective implementation steps are as follows: ①Take 12 mice and divide them into 4 groups randomly, 3 in each group; ②Take one of them The mice in the group were used as the blank control group. After the mice were killed by neck dislocation, a pair of superficial cervical lymph nodes, deep cervical lymph nodes, left axillary lymph nodes, and right axillary lymph nodes were removed from the mice, and the tissue homogenate was broken. The light absorption value of each sample at 650 nm was detected by a functional microplate reader; ③The other three groups of mice were injected with 0.2 mL of CNP, and a group of mice were sacrificed at 12 hours, 24 hours and 48 hours, and the small A pair of superficial cervical lymph nodes, deep cervical lym...

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Abstract

The invention belongs to the technical field of nanoprobes, and specifically relates to a method for tracing deep cervical lymph nodes and meningeal lymphatic vessels using nanotechnology. The method is constructed with polyglycolide lactide and pluronic F127 as main raw materials The resulting core-shell nanoparticles are used as a model nano drug-loading system (particle size 20-50 nm), and the near-infrared fluorescent dye Cy5 is loaded to form a nanoprobe. After the nanoprobe was injected subcutaneously in the right neck, the nanoprobe was enriched to the superficial cervical lymph nodes, deep cervical lymph nodes and axillary lymph nodes through the local lymphatic drainage of the tissue, and the deep cervical lymph nodes and meningeal lymphatic vessels directly connected. Therefore, nanoprobes can be rapidly transported to the meningeal lymphatic vessels for real-time imaging and tracing of deep cervical lymph nodes and meningeal lymphatic vessels, providing a basis for research on the lymphatic transport of nanoparticles in the neck and the physiology and pathology of the brain lymphatic system. Simple and direct means of detection.

Description

technical field [0001] The invention relates to the technical field of nanoprobes, in particular to a method for tracing cervical deep lymph nodes and meningeal lymphatic vessels by using nanoprobes. Background technique [0002] The lymphatic system is another set of liquid reticular circulatory system independent of the blood circulatory system, including lymphatic tissue, lymphatic vessels and lymph fluid. meaning. Studies have shown that the lymphatic system has an important impact on the occurrence and development of diseases. For example, malignant tumors often infiltrate into the lymph nodes surrounding the primary site before metastasizing to the tissue. Therefore, it is particularly important to prevent and treat diseases by developing and imaging the lymphatic system to study the structure and physiological functions of the lymphatic system. [0003] The research on the lymphatic system of the human body has been relatively thorough, but the research on the lymp...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K49/00
CPCA61K49/0032A61K49/0034A61K49/0054A61K49/0067A61K49/0093
Inventor 赵鹏飞陈永明刘立新
Owner SUN YAT SEN UNIV
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