Ultrasonic destruction of albumin microbubbles enhances gene transfection and expression in cardiac myocytes

• Published in: Chinese medical journal Vol. 124; no. 9; pp. 1395 - 1400
• Main Authors: Wang, Guo-zhong, Liu, Jing-hua, Lü, Shu-zheng, Lü, Yun, Guo, Cheng-jun, Zhao, Dong-hui, Fang, Dong-ping, He, Dong-fang, Zhou, Yuan, Ge, Chang-jiang
• Format: Journal Article
• Language: English
• Published: China Department of Cardiology, Beijing Anzhen Hospital, Capital Medical University, Beijing Institute of Heart Lung and Vessel Diseases, Beijing 100029, China 01-05-2011
• Subjects: Albumins; Animals; beta-Galactosidase - genetics; beta-Galactosidase - metabolism; Cell Survival - genetics; Cell Survival - physiology; Cells, Cultured; Microbubbles; Myocytes, Cardiac - cytology; Myocytes, Cardiac - metabolism; Rats; Rats, Wistar; Transfection - methods; Ultrasonics - methods; 半乳糖苷酶; 基因转染; 微泡; 心肌细胞; 白蛋白; 磷酸钙沉淀; 超声波; 转基因表达; cardiac myocytes; microbubbles; gene transfer; ultrasound
• ISSN: 0366-6999; 2542-5641
• DOI: 10.3760/cma.j.issn.0366-6999.2011.09.022

Background It has been proven that ultrasonic destruction of microbubbles can enhance gene transfection efficiency into the noncardiac cells, but there are few reports about cardiac myocytes. Moreover, the exact mechanisms are not yet clear; whether the characteristic of microbubbles can affect the gene transfection efficiency or not is still controversial.This study was designed to investigate whether the ultrasound destruction of gene-loaded microbubbles could enhance the plasmids carried reporter gene transfection in primary cultured myocardial cell, and evaluate the effects of microbubbles characteristics on the transgene expression in cardiac myocytes.Methods The β-galactosidase plasmids attached to the two types of microbubbles, air-contained sonicated dextrose albumin （ASDA） and perfluoropropane-exposed sonicated dextrose albumin （PESDA） were prepared. The gene transfection into cardiac myocytes was performed in vitro by naked plasmids, ultrasound exposure, ultrasonic destruction of gene-loaded microbubbles and calcium phosphate precipitation, and then the gene expression and cell viability were analyzed.Results The ultrasonic destruction of gene-loaded microbubbles enhanced gene expression in cardiac myocytes compared with naked plasmid transfection （（51.95±2..41） U/g or （29.28±3.65） U/g vs. （0.84-0.21） U/g, P ＜0.01）, and ultrasonic destruction PESDA resulted in more significant gene expression than ASDA （（51.95e2.41） U/g vs. （29.28±3.65）U/g, P ＜0.05）. Ultrasonic destruction of microbubbles during calcium phosphate precipitation gene transfection enhanced 3-galactosidase activity nearly 8-fold compared with calcium phosphate precipitation gene transfection alone （（111.35±11.21） U/g protein vs. （14.13±2.58） U/g protein, P＜0.01）. Even 6 hours after calcium phosphate precipitation gene transfection, ultrasound-mediated microbubbles destruction resulted in more intense gene expression （（35.63±7.65）U/g vs. （14.13±2.58） U/g, P＜0.05 ）.Conclusions Ultrasonic destruction of microbubbles might be a promising method for the delivery of non-viral DNA into cardiac myocytes, and the gene tranfection is related to the characteristics of microbubbles.