Yao Cuiping, Rahmanzadeh Ramtin, Endl Elmar, Zhang Zhenxi, Gerdes Johannes, Hüttmann Gereon
Key Laboratory of Biomedical Information Engineering of Ministry of Education, China.
J Biomed Opt. 2005 Nov-Dec;10(6):064012. doi: 10.1117/1.2137321.
Irradiation of nanoabsorbers with pico- and nanosecond laser pulses could result in thermal effects with a spatial confinement of less than 50 nm. Therefore absorbing nanoparticles could be used to create controlled cellular effects. We describe a combination of laser irradiation with nanoparticles, which changes the plasma membrane permeability. We demonstrate that the system enables molecules to penetrate impermeable cell membranes. Laser light at 532 nm is used to irradiate conjugates of colloidal gold, which are delivered by antibodies to the plasma membrane of the Hodgkin's disease cell line L428 and/or the human large-cell anaplastic lymphoma cell line Karpas 299. After irradiation, membrane permeability is evaluated by fluorescence microscopy and flow cytometry using propidium iodide (PI) and fluorescein isothiocyanate (FITC) dextran. The fraction of transiently permeabilized and then resealed cells is affected by the laser parameter, the gold concentration, and the membrane protein of the different cell lines to which the nanoparticles are bound. Furthermore, a dependence on particle size is found for these interactions in the different cell lines. The results suggest that after optimization, this method could be used for gene transfection and gene therapy.
用皮秒和纳秒激光脉冲照射纳米吸收体可能会产生空间限制小于50纳米的热效应。因此，吸收性纳米颗粒可用于产生可控的细胞效应。我们描述了一种激光照射与纳米颗粒相结合的方法，这种方法会改变质膜通透性。我们证明该系统能使分子穿透不可渗透的细胞膜。使用532纳米的激光照射胶体金偶联物，这些偶联物通过抗体递送至霍奇金病细胞系L428和/或人大细胞间变性淋巴瘤细胞系Karpas 299的质膜。照射后，使用碘化丙啶（PI）和异硫氰酸荧光素（FITC）葡聚糖通过荧光显微镜和流式细胞术评估膜通透性。瞬时通透然后重新封闭的细胞比例受激光参数、金浓度以及纳米颗粒所结合的不同细胞系的膜蛋白影响。此外，在不同细胞系中发现这些相互作用对颗粒大小存在依赖性。结果表明，经过优化后，该方法可用于基因转染和基因治疗。
