Krawinkel Judith, Torres-Mapa Maria Leilani, Werelius Kristian, Heisterkamp Alexander, Rüttermann Stefan, Romanos Georgios E, Gerhardt-Szép Susanne
Institute of Applied Optics, Friedrich-Schiller-University Jena, Fröbelstieg 1, Jena 07743, Germany.
Institute of Quantum Optics, Gottfried Wilhelm Leibniz University Hannover, Welfengarten 1, Hannover 30167, Germany.
Materials (Basel). 2016 May 20;9(5):397. doi: 10.3390/ma9050397.
Interaction of gold nanoparticles (AuNPs) in the vicinity of cells' membrane with a pulsed laser (λ = 532 nm, τ = 1 ns) leads to perforation of the cell membrane, thereby allowing extracellular molecules to diffuse into the cell. The objective of this study was to develop an experimental setting to deliver molecules into primary human gingival fibroblasts (pHFIB-G) by using ns-laser pulses interacting with AuNPs (study group). To compare the parameters required for manipulation of pHFIB-G with those needed for cell lines, a canine pleomorphic adenoma cell line (ZMTH3) was used (control group). Non-laser-treated cells incubated with AuNPs and the delivery molecules served as negative control. Laser irradiation (up to 35 mJ/cm²) resulted in a significant proportion of manipulated fibroblasts (up to 85%, compared to non-irradiated cells: < 0.05), while cell viability (97%) was not reduced significantly. pHFIB-G were perforated as efficiently as ZMTH3. No significant decrease of metabolic cell activity was observed up to 72 h after laser treatment. The fibroblasts took up dextrans with molecular weights up to 500 kDa. Interaction of AuNPs and a pulsed laser beam yields a spatially selective technique for manipulation of even primary cells such as pHFIB-G in high throughput.
细胞膜附近的金纳米颗粒（AuNPs）与脉冲激光（λ = 532 nm，τ = 1 ns）相互作用会导致细胞膜穿孔，从而使细胞外分子扩散进入细胞。本研究的目的是开发一种实验装置，通过使用与AuNPs相互作用的纳秒激光脉冲将分子递送至原代人牙龈成纤维细胞（pHFIB-G）（研究组）。为了比较操纵pHFIB-G与细胞系所需的参数，使用了犬多形性腺瘤细胞系（ZMTH3）（对照组）。用AuNPs和递送分子孵育的未激光处理的细胞作为阴性对照。激光照射（高达35 mJ/cm²）导致相当比例的成纤维细胞被操纵（高达85%，与未照射细胞相比：<0.05），而细胞活力（97%）没有显著降低。pHFIB-G与ZMTH3一样有效地被穿孔。激光处理后72小时内未观察到代谢细胞活性的显著下降。成纤维细胞摄取了分子量高达500 kDa的葡聚糖。AuNPs与脉冲激光束的相互作用产生了一种空间选择性技术，可用于高通量操纵甚至原代细胞，如pHFIB-G。
