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超声靶向破坏微泡增强体内兔跟腱中裸露质粒 DNA 的转染。

Ultrasound-targeted microbubble destruction enhances naked plasmid DNA transfection in rabbit Achilles tendons in vivo.

机构信息

Department of Ultrasound, West China Hospital of Sichuan University, Chengdu, China.

出版信息

Gene Ther. 2012 Jul;19(7):703-10. doi: 10.1038/gt.2011.165. Epub 2011 Oct 27.

Abstract

The study was to investigate the probability of increasing the transfection of the gene in tendons by ultrasound-targeted microbubble destruction (UTMD), and to search for the most suitable transfection conditions. A mixture of microbubbles and enhanced green fluorescent protein (EGFP) plasmids was injected into rabbit Achilles tendons by different administration routes and the tendons were ultrasound pulse by different ultrasonic conditions in order to determine the most appropriate conditions. Then, the rabbits were divided into four groups: (1) ultrasound + microbubbles + plasmid; (2) ultrasound+ plasmid; (3) microbubble + plasmid; (4) plasmid only. EGFP expression in the tendons and other tissues, and the damage to tendon and paratenon were all observed. The results showed that EGFP expression in the tendon was higher by ultrasound pulse with 2 W cm(-2) of output intensity and a 20% duty cycle for 10 min. Local injection was determined to be the better administration route. Among the four groups, EGFP expression in Group 1 was higher than that in other groups. EGFP expression was highest on seventh day, then it gradually decrease over time, and lasted more than 56 days. EGFP expression was not found in other tissues. There was no obvious injury caused by UTMD. Under suitable conditions, it is feasible to use UTMD as a safe and effective gene transfection therapy for tendon injuries.

摘要

该研究旨在探讨超声靶向微泡破坏(UTMD)是否能提高肌腱内基因转染的概率,并寻找最适合的转染条件。通过不同的给药途径将微泡和增强型绿色荧光蛋白(EGFP)质粒的混合物注入兔跟腱中,并采用不同的超声条件对肌腱进行超声脉冲,以确定最适合的条件。然后,将兔子分为四组:(1)超声+微泡+质粒;(2)超声+质粒;(3)微泡+质粒;(4)仅质粒。观察肌腱和其他组织中的 EGFP 表达情况,以及肌腱和腱旁组织的损伤情况。结果表明,输出强度为 2 W/cm²、占空比为 20%、持续时间为 10 min 的超声脉冲可使肌腱中的 EGFP 表达更高。局部注射被确定为更好的给药途径。在四组中,第 1 组的 EGFP 表达高于其他组。EGFP 表达在第 7 天最高,随后逐渐下降,持续时间超过 56 天。在其他组织中未发现 EGFP 表达。UTMD 未引起明显的组织损伤。在合适的条件下,UTMD 作为一种安全有效的基因转染治疗肌腱损伤的方法是可行的。

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