Department of Ultrasound, The Fourth Affiliated Hospital of Nantong University (The First People's Hospital of Yancheng), Jiangsu Province, China.
Bosn J Basic Med Sci. 2011 Aug;11(3):147-52. doi: 10.17305/bjbms.2011.2565.
Ultrasound-targeted microbubble destruction(UTMD) has been utilized to deliver naked siRNA into cells in in vitro settings. But whether UTMD can safely deliver naked siRNA into in vivo cells have remained unknown. This work was performed to investigate the feasibility of UTMD-enhanced naked siRNA transduction (or combined with Lipofectamine 2000) in vivo retinal cells and compare the performance between UTMD and ultrasonic irradiation alone in this enhancing effect. A dose of Cy3-labeled siRNA was injected into the vitreous cavity of rat eyes under the different conditions of Lipofectamine 2000 or/and UTMD. Transduction efficiency was assessed by fluorescence microscopy and flow cytometry. Cell and tissue damage was assessed by trypan blue exclusion test and hematoxylineosin staining, respectively. The quantity and the density of transducted cells in the group received Lipofectamine 2000 and UTMD was far more than that in other groups. The number of transducted cells in the group received Lipofectamine 2000 and ultrasonic irradiation alone was slightly more than that in the group received Lipofectamine 2000. Cy3-siRNA-positive cells can also seen in the group received UTMD alone, although the transduction efficiency is extremely low. Cell viability in each group was more than 90%, and retinal architecture in each group was well preserved. These results indicated that UTMD, with a significantly higher performance than ultrasonic irradiation alone, can effectively enhance the Lipofectamine 2000-mediated naked siRNA transduction in vivo reinal cells without any cell or tissue damage. This method can serve as a novel approach to treat the diseases of eye ground.
超声靶向微泡破坏（UTMD）已被用于将裸 siRNA 递送到体外细胞中。但 UTMD 是否能安全地将裸 siRNA 递送到体内细胞中尚不清楚。本研究旨在探讨 UTMD 增强裸 siRNA 转导（或与 Lipofectamine 2000 联合）在体内视网膜细胞中的可行性，并比较 UTMD 和单独超声辐射在这种增强效果中的性能。在不同的 Lipofectamine 2000 或/和 UTMD 条件下，将一定剂量的 Cy3 标记的 siRNA 注射到大鼠眼玻璃体腔中。通过荧光显微镜和流式细胞术评估转导效率。通过台盼蓝排斥试验和苏木精-伊红染色分别评估细胞和组织损伤。接受 Lipofectamine 2000 和 UTMD 的组中转导细胞的数量和密度远远高于其他组。接受 Lipofectamine 2000 和单独超声辐射的组中转导细胞的数量略高于接受 Lipofectamine 2000 的组。虽然转导效率极低，但在接受 UTMD 单独治疗的组中也可以看到 Cy3-siRNA 阳性细胞。各组细胞活力均超过 90%，各组视网膜结构均得到良好保存。这些结果表明，UTMD 可显著提高性能，优于单独超声辐射，可有效增强体内视网膜细胞中 Lipofectamine 2000 介导的裸 siRNA 转导，而不会造成任何细胞或组织损伤。该方法可作为治疗眼底疾病的新方法。
