Platelet membrane bionic ultrasonic microbubble as well as preparation method and application thereof

A technology of ultrasonic microbubbles and platelets, which is applied in the fields of ultrasonic molecular imaging and biomedical engineering, can solve the problems of difficulty in detection, limitation of early diagnosis of SAKI, and insensitivity to changes in hysteresis urine volume, so as to achieve more retention, less impact, and good The effect of stability

Pending Publication Date: 2021-12-10
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

However, the uncertainty and hysteresis of the creatinine baseline value, insensitivity to changes in urine volume, and difficulty in detection greatly limit the early diagnosis of SAKI, and it is difficult to provide more accurate reference results for the early diagnosis of SAKI
As for late diagnosis, the delay in diagnosis time has caused patients to miss the best treatment period, and it is still difficult to avoid adverse outcomes.
That is, creatinine and urine volume in the prior art still have certain defects as diagnostic criteria, which is not conducive to the early diagnosis of SAKI
In addition, some existing technologies also study markers for the diagnosis of SAKI, including urinary microglobulin, urinary N-acetyl-'D-glucosamine, albumin, etc., but such markers There are also their own defects, such as: easily affected by confounding factors such as inflammatory state and nutritional level, it is still difficult to apply and realize effective early diagnosis of SAKI

Method used

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  • Platelet membrane bionic ultrasonic microbubble as well as preparation method and application thereof
  • Platelet membrane bionic ultrasonic microbubble as well as preparation method and application thereof
  • Platelet membrane bionic ultrasonic microbubble as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] (1) Extraction of platelet membrane

[0049] Whole blood was collected from SD rats by abdominal aortic puncture, and ethylenediaminediaminetetraacetic acid (EDTA) was used as an anticoagulant. Specifically, 10 mM EDTA was used for anticoagulation, the whole blood was centrifuged at 22°C (without braking) at 100 g for 20 min to remove red blood cells and white blood cells, and the obtained platelet-rich plasma PRP was centrifuged at 100 g for 20 min to remove residual red blood cells.

[0050] Add prostaglandin E1PGE1 at a final concentration of 1 μM to the purified PRP supernatant to inhibit platelet activation, and centrifuge at 800 g for 20 min. The precipitate obtained after centrifugation is platelets.

[0051] After the platelets were obtained, in this embodiment, the platelet membrane and organelles were further separated by repeated freezing and thawing. Platelets were resuspended in PBS containing protease inhibitors, quickly frozen in liquid nitrogen, and dis...

Embodiment 2

[0059] Characterization of Hybrid Platelet Membrane Biomimetic Ultrasonic Microbubbles Pla-MBs

[0060] (1) Scanning electron microscope (SEM) was used to observe the surface morphology and size of microbubbles, and transmission electron microscope (TEM) was used to observe the internal structure and size of microbubbles.

[0061] TEM results such as figure 1 As shown in a, both Pla-MBs and Con-MBs have a spherical structure, but the diameter of Pla-MBs is larger and the interior is more uneven, which may be the result of platelet membrane coating. In the figure, the representative diameter of Pla-MBs microbubbles is 0.3-3.5 μm.

[0062] SEM results such as figure 1 As shown in b, after the platelet membrane hybridization, the size of Pla-MBs is slightly larger and the surface is more heterogeneous than Con-MBs, which is consistent with the TEM results.

[0063] (2) Using a fluorescence microscope to observe the fusion of artificial phospholipids and platelet membranes on t...

Embodiment 3

[0076] In vitro stability, imaging ability, safety and platelet-mimetic function of platelet membrane ultrasound microbubbles Pla-MBs

[0077] (1) Detection of the stability and imaging ability of Pla-MBs obtained by mixing different proportions of platelet membrane and phospholipids

[0078] In order to evaluate the appropriate mixing ratio of platelet membrane and phospholipid, different ratios of platelet membrane and phospholipid were set, prepared according to the steps in Example 1, and bionic ultrasonic microbubbles of platelet membrane at different mixing ratios were obtained, and Pla-MBs ultrasonic imaging was observed in vitro. Specifically, for the platelet membrane biomimetic ultrasonic microbubbles in different mixing ratios, the same steps as below are taken for imaging detection.

[0079] Add 1ml platelet membrane bionic ultrasonic microbubble diluent into 2% agarose model, use small animal ultrasonic imaging system (MX250 high-frequency linear array probe, nonl...

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Abstract

The invention discloses a platelet membrane bionic ultrasonic microbubble. The platelet membrane bionic ultrasonic microbubble comprises a shell membrane material and filling gas; the shell membrane material comprises phospholipid and a platelet membrane; and the platelet membrane covers the phospholipid and / or is dispersed in the phospholipid. The platelet membrane bionic ultrasonic microbubble has good stability, ultrasonic imaging ability, cytocompatibility and collagen targeting in vitro, and the platelet membrane bionic ultrasonic microbubble has long in-vivo circulation time and can be remarkably retained in the kidney; and based on platelet membrane functional protein modification, ultrasonic targeted imaging and directional microbubble destruction technologies, detection of an early vascular injury adhesion signal of the sepsis renal injury can be realized, and early diagnosis of the sepsis renal injury is realized. Based on the platelet membrane bionic ultrasonic microbubble provided by the invention, a simple and convenient sepsis renal injury monitoring method can be provided in combination with ultrasonic equipment, and early diagnosis can be realized, so that the defects of diagnostic standards in the prior art can be overcome, and a patient can be correspondingly treated as soon as possible, thereby obtaining a good clinical result.

Description

technical field [0001] The invention relates to the fields of ultrasonic molecular imaging and biomedical engineering, and more specifically relates to a platelet membrane bionic ultrasonic microbubble and its preparation method and application. Background technique [0002] Acute kidney injury (acute kidney injury, AKI) refers to a short period of time, due to various reasons, the glomerular filtration rate suddenly drops, leading to the accumulation of nitrogenous substances, which leads to the rapid development of water, electrolytes, acid-base balance and systemic complications. sign. Sepsis and septic shock are the main causes of AKI in critically ill patients. The survey shows that more than 50% of AKI in ICU is related to sepsis. At the same time, sepsis-induced acute kidney injury (SAKI) is different from non-sepsis-induced AKI and has a unique pathophysiological mechanism. Great progress has been made, but it is still unable to meet the actual treatment needs, and...

Claims

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Application Information

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Patent Type & AuthorityApplications(China)
IPC IPC(8): A61K49/22
CPCA61K49/223
Inventor杨静缪小燕关宇陈素芳张政尹庭辉姚伟锋黑子清
OwnerTHE THIRD AFFILIATED HOSPITAL OF SUN YAT SEN UNIV