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Bionic lipid microbubble ultrasonic contrast agent constructed by cell membranes and preparation method of contrast agent

An ultrasonic contrast agent and microbubble technology, which is applied in the fields of ultrasonic molecular imaging and biomedical engineering, can solve the problems of poor stability and lack of natural targeting of microbubble ultrasonic contrast agents, and achieve the effect of overcoming low biocompatibility

Active Publication Date: 2020-06-26
THE THIRD AFFILIATED HOSPITAL OF SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] One of the objectives of the present invention is to provide a bionic microbubble ultrasound contrast agent for the defects of lack of natural targeting and poor stability of existing microbubble ultrasound contrast agents

Method used

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  • Bionic lipid microbubble ultrasonic contrast agent constructed by cell membranes and preparation method of contrast agent
  • Bionic lipid microbubble ultrasonic contrast agent constructed by cell membranes and preparation method of contrast agent
  • Bionic lipid microbubble ultrasonic contrast agent constructed by cell membranes and preparation method of contrast agent

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0067] Example 1: Preparation of ultrasound contrast agent imitating leukocyte microbubbles

[0068] Rats were intraperitoneally injected with LPS 1.5 mg / kg. Six hours later, after sevoflurane inhalation anesthesia, the abdominal cavity was opened to expose the abdominal aorta to collect blood in anticoagulant blood collection tubes. Take another 50ml centrifuge tube and add the same volume of rat peripheral blood leukocyte separation medium (Solarbio) as the blood sample; use a Pasteur pipette to draw the blood sample, spread it carefully on the surface of the separation medium, and centrifuge at 1000g for 30min. After centrifugation, two ring-shaped milky white cell layers will appear in the centrifuge tube, the upper layer of cells is a mononuclear cell layer, and the lower layer of cells is a white blood cell layer. Use a pipette to carefully absorb the white blood cell layer in the separation solution, add 3 times the cell volume of red blood cell lysate (Solarbio), gentl...

Embodiment 2

[0076] Example 2: Detection of Biomimetic Microvesicle Targeting Ligands

[0077] The control and biomimetic microbubbles prepared above were purified by centrifugation (2000 rpm, 2 min), the lower layer liquid was discarded, resuspended in 500 μl PBS, and 4.5 μg CD45 flow cytometry antibody was added, and incubated at room temperature for 30 minutes. Then centrifuge again (2000rpm, 2min), discard the lower liquid to remove free antibody, add 500μl PBS to resuspend, dilute to 10 5 cells / ml, the expression of microvesicle CD45 was detected by flow cytometry.

[0078] The results showed that: biomimetic microbubbles (MB m ) CD45 expression level (30.6%) was significantly higher than control microvesicles (MB con )(3.34%)( image 3 ), indicating that the biomimetic microvesicles carry the leukocyte membrane-specific protein CD45.

[0079] The control and biomimetic microbubbles prepared above were purified by centrifugation (2000 rpm, 2 min), the lower layer liquid was discar...

Embodiment 3

[0082] Example 3: Ultrasound imaging ability detection of bionic microbubble ultrasound contrast agent

[0083] Establish rat liver ischemia-reperfusion injury (ischemia-reperfusion injury, IRI) model: rats were fasted for 8-12 hours before surgery, and anesthetized by intraperitoneal injection of a compound solution of ketamine (60mg / kg) and xylazine (100mg / kg) . Fix the rat on the operating table, make a median abdominal incision, expose the abdominal cavity, dissociate the hilar blood vessels, clamp the blood flow in the left lobe and middle lobe of the liver with blood vessel clips, remove the blood vessel clips after 60 min, and suture the incision. The operation of ischemia 0min in the control group was performed after laparotomy.

[0084] After the animals were anesthetized, they were fixed on the operating table. Cut the suture along the midline abdominal incision, and fix the ultrasound probe (color Doppler ultrasound diagnostic system, Philips EPQ7, 5-12L high-freq...

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Abstract

The invention belongs to the field of medicines, and relates to a bionic lipid microbubble ultrasonic contrast agent constructed by cell membranes and a preparation method of the contrast agent. The preparation method comprises the following steps: firstly, separating cells; extracting cell membranes; ultrasonically vibrating the cell membranes and phospholipids to prepare liposomes of mixed cellmembranes and phospholipids; and finally, introducing gas into the liposomes, and performing mechanical vibration to prepare the targeted microbubble ultrasonic contrast agent. According to the bioniclipid microbubble ultrasound contrast agent disclosed by the invention, the biological characteristics of the cell membranes are integrated into the ultrasound contrast agent, so that the applicationrange of the traditional microbubble ultrasound contrast agent is broken through. The preparation method of the bionic liposome has the advantages of natural targeting and easy clinical transformation.

Description

technical field [0001] The invention relates to the fields of ultrasonic molecular imaging and biomedical engineering, in particular to a bionic microbubble ultrasonic contrast agent and a preparation method thereof. [0002] technical background [0003] Ultrasound molecular imaging is a molecular imaging method that uses targeted microbubble contrast agents as molecular probes, and uses contrast-enhanced ultrasound as an imaging mode to conduct qualitative and quantitative research on biochemical processes in vivo at the cellular and molecular levels, which can improve the diagnostic accuracy of ultrasound. Accuracy and sensitivity have important application prospects in the fields of cardiovascular, inflammatory, tumor and other diseases. [0004] The advantage of ultrasound imaging is that its operation is simple and fast, and the dynamic development of vascular injury lesions can be observed in real time, which is economical and reduces the economic pressure of patients....

Claims

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Application Information

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IPC IPC(8): A61K49/22
CPCA61K49/223
Inventor 尹庭辉张政缪小燕游宇佳郑荣琴周慧超任杰邱晨姚伟锋杨静黑子清
Owner THE THIRD AFFILIATED HOSPITAL OF SUN YAT SEN UNIV
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