A study of the ultrasound-targeted microbubble destruction based triplex-forming oligodexinucleotide delivery system to inhibit tissue factor expression

Liang,Weihua; Zhang,Weiwei; Zhao,Shifu; Li,Qianning; Yang,Yiming; Liang,Hua; Ceng,Rongchuan

doi:10.3892/mmr.2014.2822

A study of the ultrasound-targeted microbubble destruction based triplex-forming oligodexinucleotide delivery system to inhibit tissue factor expression

Authors:
- Weihua Liang
- Weiwei Zhang
- Shifu Zhao
- Qianning Li
- Yiming Yang
- Hua Liang
- Rongchuan Ceng
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Affiliations: Department of Neurology, Xinqiao Hospital, The Third Military Medical University, Chongqing 400038, P.R. China, Deparment of Neurology, General Hospital of Beijing PLA Military Region, Beijing 100700, P.R. China, Department of Internal Medicine, The Sixteenth Hospital of PLA, Altay, Xinjiang 836500, P.R. China, Department of Internal Medicine, 66083 Clinic of Beijing Military Region, Beijing 102488, P.R. China
Published online on: October 30, 2014 https://doi.org/10.3892/mmr.2014.2822
Pages: 903-909
Copyright: © Liang et al. This is an open access article distributed under the terms of Creative Commons Attribution License [CC BY_NC 3.0].

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Abstract

The efficiency of cellular uptake of triplex‑forming oligodexinucleotides (TFO), and the inhibition of tissue factor (TF) is low. The aim of the present study was to improve the absorption of TFO, and increase the inhibition of TF induced by shear stress both in vitro and in vivo, by using an ultrasound‑targeted microbubble destruction (UTMD)‑based delivery system. TFO‑conjugated lipid ultrasonic microbubbles (TFO‑M) were first constructed and characterised. The absorption of TFO was observed by a fluorescence‑based method, and the inhibition of TF by immunofluorescence and quantitative polymerase chain reaction. ECV304 human umbilical vein endothelial cells were subjected to fluid shear stress for 6 h after treatment with TFO conjugated lipid ultrasonic microbubbles without sonication (TFO‑M group); TFO alone; TFO conjugated lipid ultrasonic microbubbles, plus immediate sonication (TFO+U group and TFO‑M+U group); or mock treated with 0.9% NaCl only (SSRE group). The in vivo experiments were established in a similar manner to the in vitro experiments, except that TFO or TFO‑M was injected into rats through the tail vein. Six hours after the preparation of a carotid stenosis model, the rats were humanely sacrificed. The transfection efficiency of TFO in the TFO‑M+U group was higher as compared with the TFO‑M and TFO+U group (P<0.01). The protein and mRNA expression of TF in the TFO‑M+U group was significantly decreased both in vitro and in vivo (P<0.01), as compared with the TFO‑M, TFO+U and SSRE groups. The UTMD‑based TFO delivery system promoted the absorption of TFO and the inhibition of TF, and was therefore considered to be favorable for preventing thrombosis induced by shear stress.

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