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Ultrasonic microbubble contrast medium containing gene

A technology of ultrasonic microbubbles and contrast agents, applied in echo/ultrasound imaging agents, gene therapy, hypodermic injection devices, etc.

Inactive Publication Date: 2006-10-18
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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  • Ultrasonic microbubble contrast medium containing gene
  • Ultrasonic microbubble contrast medium containing gene
  • Ultrasonic microbubble contrast medium containing gene

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0022] Example 1: The present invention is used for the synthesis of the self-made contrast agent of the encapsulation of gene

[0023] Lecithin, cholesterol, polyethylene glycol stearyl ethanolamine were dissolved in chloroform at a mass ratio of 1:3:3, and evaporated in a rotary vacuum to form a film; 0.9% sodium chloride solution, propylene glycol and glycerin (0.9% chlorine Sodium chloride solution: propylene glycol: glycerin = 8:1:1), shake to wash the membrane 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 microbub...

example 2

[0024] Example 2: Detection of Gene Adherence to Microbubble Contrast Agents

[0025] 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

[0026] Example 3: Synthesis of gene encapsulation in microbubble contrast agent

[0027] 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 in chloroform according to the mass ratio of 1:3:3, and evaporate in a rotary vacuum to form a film; add an appropriate amount of aqueous solvent: 0.9% sodium chloride solution; propylene glycol; glycerol=8:1:1 and shake the film to form liposomes Suspension; freeze overnight; after thawing, vibrate for 120 seconds at 35% of the maximum output power with a sonicator, and at the same time slowly pour about 6ml of the mixture of fluorocarbon liquid and gene into the bottom through the three-way tube to form lipids Somatic fluorocarbon contrast agent (the outer shell is liposome, and the inner package is solid fluorocarbon liquid and selected genes). Wash...

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Abstract

The invention relates to a method for transfecting genes in targeted tissues by using ultrasonic microbubble contrast agents as gene delivery vehicles for positioning and controlled release of genes. The gene-carrying ultrasonic microbubble contrast agent includes the situation that the gene is adhered to the surface of the contrast agent microbubble and / or the gene is encapsulated in the microbubble contrast agent. For intravenously injected microbubble contrast agents or gene-carrying microbubble contrast agents, use sonograms to monitor the arrival of the contrast agents to the target tissue. Monitor gene transfection and expression. The invention has the advantages of non-invasiveness, simple operation, good targeting, high transfection rate, safety, repeated use and the like.

Description

technical field [0001] The invention relates to an ultrasonic microbubble contrast agent as a gene carrier, in particular to an ultrasonic microbubble contrast agent carrying a gene and a preparation method thereof. The contrast agent can be guided by the sonogram, and the gene transfection can be completed by localizing and triggering the ultrasound with a certain energy. 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. [0003] Gene transfection refers to the process of introducing phage, virus or recombinants constructed with them into cells. At present, common gene transfer methods include virus-mediated ge...

Claims

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Application Information

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