Walter-Brendel-Centre of Experimental Medicine, Schillerstrasse 44, 80336 Munich, Germany.
Pharm Res. 2012 May;29(5):1282-94. doi: 10.1007/s11095-012-0678-8. Epub 2012 Jan 25.
Site specific vascular gene delivery is a promising tool for treatment of cardiovascular diseases. By combining ultrasound mediated microbubble destruction with site specific magnetic targeting of lentiviruses, we aimed to develop a technique suitable for systemic application.
The magnetic nanoparticle coupling to lipid microbubbles was confirmed by absorbance measurements. Association of fluorescent lentivirus to magnetic microbubbles (MMB) was determined by microscopy and flow cytometry. Functionality and efficiency of GFP-encoding lentiviral MMB transduction was evaluated by endothelial (HMEC) GFP expression and cytotoxicity was measured by MTT reduction.
Microbubbles with a mean diameter of 4.3 ± 0.04 μm were stable for 2 days, readily magnetizable and magnetically steerable in vitro and efficiently associated with lentivirus. Exposure of eGFP-encoding lentiviral MMB to human endothelial cells followed by application of an external static magnetic field (30 min) and ultrasonic destruction of the microbubbles did not markedly affect cellular viability. Finally, this combination led to a 30-fold increase in transduction efficiency compared to application of naked virus alone.
By associating microbubbles with magnetic iron nanoparticles, these function as carriers for lentiviruses achieving tissue specific deposition at the site of interest.
针对特定部位的血管基因传递是治疗心血管疾病的一种很有前途的工具。通过将超声介导的微泡破坏与慢病毒的特定部位磁靶向相结合，我们旨在开发一种适合系统应用的技术。
通过吸光度测量来确认磁性纳米颗粒与脂质微泡的结合。通过显微镜和流式细胞术确定荧光慢病毒与磁性微泡（MMB）的结合。通过内皮细胞（HMEC）GFP 表达评估 GFP 编码慢病毒 MMB 转导的功能和效率，并通过 MTT 减少测量细胞毒性。
平均直径为 4.3±0.04μm 的微泡稳定 2 天，易于磁化，在体外可磁引导，并且与慢病毒有效结合。将 eGFP 编码慢病毒 MMB 暴露于人内皮细胞，然后施加外部静态磁场（30 分钟）并超声破坏微泡，不会明显影响细胞活力。最后，与单独应用裸病毒相比，这种组合使转导效率提高了 30 倍。
通过将微泡与磁性铁纳米颗粒结合，这些颗粒可以作为慢病毒的载体，实现在感兴趣部位的组织特异性沉积。
