A targeted ultrasound contrast agent carrying gene and cell-penetrating peptide: Preparation and gene transfection in vitro

A thrombus-targeted ultrasound contrast agent bound with tirofiban — a glycoprotein (GP) IIb/IIIa antagonist that can specifically bind to activated platelets in the thrombus — was designed to enhance both the image contrast and thrombolysis effect. In this study, we used 76 canine thrombi for investigation. The targeting ability to thrombi was confirmed by microphotography and high-frequency ultrasound (40 MHz) imaging. The effect of the targeted microbubbles on thrombolysis enhancement was investigated using an in vitro flow system: targeted and nontargeted microbubbles flowed through the clot for 30 seconds with a washing step; the microbubbles remained on the clot that were then cavitated by ultrasound (frequency = 1 MHz, MI = 1.2). The extent of thrombolysis was evaluated by weight reduction and histology analysis. The targeted microbubbles reduced the weight of thrombi by a factor of 1.7 times that of the nontargeted microbubbles. (clot weight reduction: 23.1 ± 5.3% versus 13.6 ± 4.9%, p < 0.01 between targeted and nontargeted group), and the signal enhancement was 3.34 ± 0.30 dB (mean ± SD, p < 0.01 compared to control). We conclude that targeted microbubbles are applicable not only for molecular imaging of thrombi but also for improving the effectiveness of ultrasound-assisted thrombolysis.

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A novel dual-targeted ultrasound contrast agent provides improvement of gene delivery efficiency in vitro

Tumor Biology ◽

10.1007/s13277-015-4681-7 ◽

2016 ◽

Vol 37 (7) ◽

pp. 8609-8619 ◽

Cited By ~ 4

Author(s):

Jinfeng Xu ◽

Xinxin Zeng ◽

Yingying Liu ◽

Hui Luo ◽

Zhanghong Wei ◽

...

Keyword(s):

Gene Delivery ◽

Contrast Agent ◽

Ultrasound Contrast Agent ◽

Ultrasound Contrast ◽

Targeted Ultrasound ◽

Delivery Efficiency

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Preparation and In Vitro Evaluation of a Nano Ultrasound Contrast Agent Targeting Pancreatic Cancer

Journal of Nanoscience and Nanotechnology ◽

10.1166/jnn.2021.18883 ◽

2021 ◽

Vol 21 (3) ◽

pp. 1413-1418

Author(s):

Wu Chen ◽

Xiaofang Liu ◽

Yiying Li ◽

Yongsheng Yang ◽

Kun Xu ◽

...

Keyword(s):

Pancreatic Cancer ◽

Particle Size ◽

Contrast Agent ◽

Drug Loading ◽

Ultrasound Contrast Agent ◽

Ultrasound Contrast ◽

Loading Efficiency ◽

Targeted Ultrasound ◽

Drug Loading Efficiency

To prepare a nano-sized ultrasound contrast agent that specifically targets pancreatic cancer cells and to evaluate its targeting effect In Vitro. PLGA-PEG-NHS was synthesized using PLGA, NHS, and PEG and detected using 1H-NMR. PLGA-PEG-NHS and PFOB were used to prepare PLGA nano contrast agent coated with PFOB by emulsification and volatilization, and then a hedgehog antibody was conjugated. The morphology of the nano contrast agent was observed using a transmission electron microscope, and its particle size and potential were measured using the dynamic light scattering method. The entrapment and drug loading efficiency of the nano contrast agent was measured using gas chromatography-mass spectrometry. The In Vitro release characteristics of the nano contrast agent was measured using the dialysis method. Human pancreatic cancer cell lines SW1990 and CFPAC1 were cultured in medium containing the nano contrast agent. The targeting ability of the nano contrast agent was qualitatively and quantitatively verified using fluorescence microscopy and flow cytometry. The average particle size of the targeted ultrasound contrast agent was 198.9 nm, zeta potential was −31.8 mv, entrapment rate was 63.7±3.9%, drug loading efficiency was 14.3±0.9%, and drug release was 85.3% in 48 h. In Vitro cell experiments showed that the targeted ultrasound contrast agent strongly bound to SW1990 cells with high expression of hedgehog antigen, but no specific binding was detected in CFPAC-1 cells which lack the hedgehog antigen. The nano ultrasound contrast agent prepared by emulsification and volatilization method can be potentially used for the diagnosis of pancreatic cancer.

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Preparation and in vitro evaluation of pancreatic cancer-targeted nano-scale ultrasound contrast agent

Academic Journal of Second Military Medical University ◽

10.3724/sp.j.1008.2015.01277 ◽

2015 ◽

Vol 36 (12) ◽

pp. 1277

Author(s):

Chu-ling HU ◽

Zhong-jian CHEN ◽

Zong-guang TAI ◽

Yuan GAO ◽

Shen GAO ◽

...

Keyword(s):

Pancreatic Cancer ◽

Contrast Agent ◽

Ultrasound Contrast Agent ◽

In Vitro Evaluation ◽

Nano Scale ◽

Ultrasound Contrast

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Novel Difolate Targeting Nano-Level Ultrasound Contrast Agent for Therapy of Breast Cancer Tumor Cells

Science of Advanced Materials ◽

10.1166/sam.2021.4038 ◽

2021 ◽

Vol 13 (7) ◽

pp. 1295-1303

Author(s):

Guangheng Liu ◽

Xiangfeng Yang ◽

Qiming Niu ◽

Wenkui Sun

Keyword(s):

Breast Cancer ◽

Contrast Agent ◽

Tumor Cells ◽

Folate Receptor ◽

Ultrasound Contrast Agent ◽

Hydroxyl Group ◽

Binding Ability ◽

Ultrasound Contrast ◽

Mcf 7

ABSTRACTA new type of difolate targeting nano-level ultrasound contrast agent ((folate molecule, FOL)2-TUAs) was prepared, so as to investigate its targeted binding effect with human breast cancer mammary carcinoma cells (MCF-7) in vitro. L-2-aminoadipic acid (L-2-AD) as a branch unit was inserted at the hydroxyl end of distearoyl phosphatidylethanolamine (DISP)-PEG2000-COOH to construct a tree structure. At this time, the free hydroxyl group in the distearoyl phosphatidylethanolamine (DISP)-PEG2000-COOH structure modified the FOL with the help of N-Hydroxysuccinimide/N,N'-dicyclohexylcarbodiimide (NHS/DCC). Each 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethylene glycol)-2000] (DISP-PEG2000) connected two FOLs to generate difolate targeted nanomaterials. Nano laser particle size (PS) and Zeta potential analyzer (ZPA) were applied to analyze the physical characteristics of the material such as PS and dispersion, and the enhanced development effect in vitro was detected by the ultrasonic diagnostic instrument. Besides, the targeted binding ability of the contrast agent based on this material to folate receptor (FR) overexpressing MCF-7 cells was analyzed by flow cytometry (FCM) and fluorescence microscope. In the experiment, hydrogen-1 nuclear magnetic resonance (1H NMR) demonstrated that (FOL)2-TUAs was successfully synthesized. The surface of this material was round and uniformly distributed without aggregation. According to the relative number of FOL molecules, non-targeted nano-agent (U-TUA), monofolate targeted nano-agent (FOL-TUA), and difolate targeted nano-agent ((FOL)2-TUA) were obtained. The in vitro imaging showed that different materials exhibited enhanced imaging effects in ultrasonic diagnostic equipment. FCM and fluorescence microscopy both indicated that the difolate TUA could achieve a good binding to MCF-7 cells. Most of the nano-agents were attached to the cell membrane, surrounded by red fluorophore, namely increasing the FOL content of DISP-PEG2000 chain could enhance the targeted binding ability of tumor cells.

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