Departments of Ophthalmology, The Second Affiliated Hospital of Chongqing Medical University, 74# Linjiang Road, Chongqing, 400010, People's Republic of China.
Lasers Med Sci. 2010 Jul;25(4):587-94. doi: 10.1007/s10103-010-0773-1. Epub 2010 Mar 20.
Laser-mediated gene transfection has received much attention as a new method for targeted gene therapy because of the high controllability of laser energy and direction. In this report, we describe a combination laser-microbubble system that enables membrane-impermeable molecules to penetrate cell membranes. The main theories we apply are optical breakdown and photoacoustic generation, which are induced by laser irradiation. Firstly, different types of laser light (Ar-green, Novus Varia poly-wavelength and Nd:YAG laser) were adopted to blast liposome microbubble contrast medium; subsequently, the Nd:YAG laser (1064 nm, 4 ns), which could successfully blast microbubbles, and ultrasound were used in combination to irradiate a mixture of liposome microbubbles and retinoblastoma (Rb) cells. After irradiation, membrane permeability was evaluated by flow cytometric assay using propidium iodide (PI) and fluorescein diacetate (FDA). The proportion of permeabilized resealed cells was affected by changes in the light energy. All of the Nd:YAG laser, Nd:YAG combination laser-microbubble and combination ultrasound-microbubble systems were able to permeabilize the Rb cells. These results suggest that this combination laser-microbubble system is a new means of delivering exogenous materials into living cells.
激光介导的基因转染作为一种新的靶向基因治疗方法受到了广泛关注，因为激光能量和方向具有高度可控性。在本报告中，我们描述了一种组合激光-微泡系统，该系统能够使细胞膜不可渗透的分子穿透细胞膜。我们应用的主要理论是由激光照射诱导的光击穿和光声产生。首先，采用不同类型的激光（Ar-绿光、Novus Varia 多波长和 Nd:YAG 激光）爆破脂质体微泡造影剂；随后，采用能够成功爆破微泡的 Nd:YAG 激光（1064nm，4ns）与超声联合照射脂质体微泡和视网膜母细胞瘤（Rb）细胞混合物。照射后，通过碘化丙啶（PI）和荧光素二乙酸（FDA）的流式细胞术检测评估细胞膜通透性。再透化细胞的比例受光能量变化的影响。所有 Nd:YAG 激光、Nd:YAG 联合激光-微泡和联合超声-微泡系统都能够使 Rb 细胞透化。这些结果表明，这种组合激光-微泡系统是将外源性物质递送入活细胞的一种新方法。
