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  • Title: Ultrasound-Targeted Microbubble Destruction Enhances Gene Expression of microRNA-21 in Swine Heart via Intracoronary Delivery.
    Author: Liu Y, Li L, Su Q, Liu T, Ma Z, Yang H.
    Journal: Echocardiography; 2015 Sep; 32(9):1407-16. PubMed ID: 25613289.
    Abstract:
    BACKGROUND: Ultrasound-targeted microbubble destruction (UTMD) has proved to be a promising method for gene delivery. However, the feasibility and efficacy of UTMD-mediated gene delivery to the heart of large animals remain unclear. The present study was to explore the probability of increasing the transfection of microRNA-21 (miR-21) in swine heart by UTMD, and to search for the most suitable transfection conditions. METHODS: We first optimized ultrasound intensity for successful miR-21 delivery. After intravenous injection of miR-21/microbubble mixture (miR-21/MB), transthoracic ultrasound irradiation (US) was applied from the left anterior chest using different intensities (1, 2, and 3 W/cm(2)). Then the efficacy of UTMD-mediated miR-21 delivery into myocardium via intracoronary injection was explored. Solution of miR-21/MB was infused intravenously or intracoronarily with US over the heart. Swine undergoing phosphate-buffered saline (PBS) injection, miR-21/MB injection via ear vein or coronary artery without US served as the control. The dynamic changes of left ventricular ejection fraction (LVEF) and serum troponin I (cTnI) after UTMD were detected, then the left ventricular myocardium was harvested for hematoxylin and eosin (H&E) staining 4 days later; the expression levels of miR-21 and programmed cell death 4 (PDCD4) were detected by quantitative real time polymerase chain reaction (qRT-PCR) and Western blot, respectively. RESULTS: Results showed that pulse ultrasound at an intensity of 2 W/cm(2) and a 50% duty ratio for 20 minutes, there was no increase in serum cTnI, no histological sign of myocardial damage, and no noted cardiac dysfunction with relatively higher miR-21 expression (P < 0.05). Compared to miR-21/MB alone, UTMD significantly increased gene expression in myocardium regardless of the delivery routes (P < 0.05). Interestingly, the transfection efficiency was found to be a little bit higher with intracoronary injection than that with intravenous injection, though the dose for intracoronary injection was half of the intravenous injection (P < 0.05). CONCLUSION: Under suitable conditions, UTMD can efficiently enhance gene expression in swine heart regardless of the delivery routes. The intravenous injection might be superior to intracoronary injection with less invasiveness and lower requirement of the technique. And for those undergoing percutaneous coronary intervention, intracoronary injection seems to be another alternative.
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