Duvshani-Eshet Maayan, Adam Dan, Machluf Marcelle
The Laboratory of Cancer Drug Delivery and Mammalian Cell Technology, Faculty of Biotechnology and Food Engineering, Technion-Israel Institute of Technology, Haifa 32000, Israel.
J Control Release. 2006 May 15;112(2):156-66. doi: 10.1016/j.jconrel.2006.02.013. Epub 2006 Feb 28.
The application of therapeutic ultrasound (TUS) in combination with contrast agents (USCA) to mediate gene delivery relies on the understanding of the bioeffects involved. The objective of this study was to evaluate the various bioeffects generated by albumin-coated microbubbles: Optison, an USCA, when applied with TUS operated for 10-30 min, on cells and on DNA transfection. This study reveals that Optison microbubbles were still acoustically active after long-term TUS application of 30 min. Optison enhances TUS-gene transfection by increasing the number of plasmids in the cells and also by distributing the plasmids to more cells, without significant decrease in cell viability. Optison also interacts with the DNA to further enhance transfection in a mechanism not necessarily involving cavitation. However, Optison affects mainly the cell cytoplasmatic membrane, without interfering with DNA intracellular trafficking. Using high-resolution scanning electron microscopy (HRSEM), the bioeffects on cell membrane induced by TUS-Optison were observed, demonstrating that Optison lead to a rougher surface, characterized by depressions that are reversible within 24-h post TUS. These effects are different from those observed when only TUS was applied. The findings from this study suggest that albumin-coated microbubbles enhances transfection when using TUS for 10-30 min, and that microbubbles play a major role in elevating cell transfection level and efficiency.
治疗性超声（TUS）联合造影剂（USCA）介导基因递送的应用依赖于对相关生物效应的理解。本研究的目的是评估白蛋白包被的微泡Optison（一种USCA）在与持续10 - 30分钟的TUS联合应用时，对细胞和DNA转染产生的各种生物效应。本研究表明，在30分钟的长期TUS应用后，Optison微泡仍具有声学活性。Optison通过增加细胞内质粒数量以及将质粒分布到更多细胞来增强TUS介导的基因转染，且细胞活力无显著下降。Optison还与DNA相互作用，以一种不一定涉及空化的机制进一步增强转染。然而，Optison主要影响细胞质膜，而不干扰DNA的细胞内运输。使用高分辨率扫描电子显微镜（HRSEM）观察了TUS - Optison对细胞膜的生物效应，结果表明Optison导致细胞膜表面更粗糙，其特征是出现凹陷，这些凹陷在TUS后24小时内可恢复。这些效应与仅应用TUS时观察到的不同。本研究结果表明，白蛋白包被的微泡在TUS持续10 - 30分钟时可增强转染，且微泡在提高细胞转染水平和效率方面起主要作用。
