Location controlled releasing method for ultrasonic microbubble contrast medium

A technology of ultrasonic microbubbles and contrast agents, applied in echo/ultrasonic imaging agents, pharmaceutical formulations, hypodermic injection devices, etc.

Inactive Publication Date: 2005-01-05
王志刚
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no study on the synthesis of gene-carrying or gene-targeted ultrasound microbubble contrast agents and their targeted release methodology, and there is no methodology for monitoring their release and expression in tissues using sonographic and tissue characterization methods

Method used

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  • Location controlled releasing method for ultrasonic microbubble contrast medium
  • Location controlled releasing method for ultrasonic microbubble contrast medium
  • Location controlled releasing method for ultrasonic microbubble contrast medium

Examples

Experimental program
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example 1

[0026] Example 1: Synthesis of Microbubble Contrast Agents of the Invention for Adhesion of Genes

[0027] Lecithin, cholesterol, polyethylene glycol stearyl ethanolamine are dissolved in chloroform in a ratio of 1:3:3 by weight and number, and evaporated in a rotary vacuum to form a film; add 0.9% sodium chloride solution, propylene glycol and glycerol (0.9 % sodium chloride solution: propylene glycol: glycerin=8:1:1), wash the membrane by shaking to form a liposome suspension. Freeze overnight. After thawing, vibrate for 80s with a vibrator at 30% of the maximum output power, and at the same time slowly fill 0.6ml of perfluoropropane gas below through a three-way tube to form lipid fluorocarbon microbubbles. Use a microporous membrane to filter out microbubbles with larger particle sizes. The concentration of lipid microbubble contrast agent was determined by Kool particle counter, and the particle size was determined by microscope. The measured microbubble size is 2-4um,...

example 2

[0028] Example 2: Detection of Gene Adherence to Microbubble Contrast Agents

[0029] The lipid shell of the microbubble contrast agent synthesized in Example 1 was fluorescently labeled, and the genes were stained with propidium iodide (PI). Mix 3ml of microbubble contrast agent with 1ml of gene at 4°C for 2 hours, and detect the number of microbubbles emitting two kinds of fluorescence by immunofluorescence method, which is the number of microbubbles adhered to the gene.

example 3

[0030] Example 3: Production of gene encapsulation in microbubble contrast agent

[0031] Mix perfluoropropane liquid and green fluorescent protein gene at a volume ratio of 3:1 at 4°C to form a mixture of fluorocarbon liquid and gene, using lipid materials lecithin, cholesterol, polyethylene glycol stearyl ethanolamine Dissolve with chloroform at a ratio of 1:3:3 by weight and number of parts, and then evaporate in a rotary vacuum to form a film; add an appropriate amount of aqueous phase solvent in proportion: the volume and number of parts is 0.9% sodium chloride solution: propylene glycol: glycerol=8:1: 1 Oscillate to wash the membrane to form a liposome suspension; freeze overnight; after thawing, vibrate with a sonicator at 35% of the maximum output power for an appropriate time of 120s, and at the same time slowly pour in the fluorocarbon liquid and About 6ml of the gene mixture forms a liposome fluorocarbon contrast agent (the outer shell is a liposome, and the solid f...

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Abstract

A method for using the ultrasonic microvesicle contrast medium as the carrying tool of gene to locate the gene and control to release it to target tissue for transfection. After the contrast medium reaches the target tissue under themonitor of audio-video image, the ultrasonic waves are used to destroy the microvesicle for releasing the gene and performing the gene transfection.

Description

technical field [0001] The invention relates to the use of ultrasonic microbubble contrast agent as a carrier tool for genes, guided by ultrasonic images, and triggered by certain energy ultrasonic positioning to carry out gene transfection, and provides a method for positioning and monitoring by using ultrasonic images and ultrasonic tissue characterization methods methods for targeted gene transfer. Background technique [0002] Ultrasonic microbubble contrast agent, as a new type of acoustic contrast agent, has played a huge role in the ultrasound imaging of organs and tissues. With the development of ultrasound technology such as second harmonic and triggered imaging and the improvement of contrast agent preparation technology, contrast agent can greatly improve the diagnosis rate of various tissue lesions, and the frequency of ultrasound routinely used for diagnosis exceeds 1MHz. [0003] Gene transfection refers to the process of introducing phage, virus or recombinan...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K48/00A61K49/22A61M5/00
Inventor 王志刚冉海涛任红凌智瑜
Owner 王志刚
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