Kulbacka Julita, Pucek Agata, Kotulska Małgorzata, Dubińska-Magiera Magda, Rossowska Joanna, Rols Marie-Pierre, Wilk Kazimiera Anna
Department of Medical Biochemistry, Medical University, Chalubinskiego 10 St., 50-368 Wrocław, Poland.
Department of Organic and Pharmaceutical Technology, Faculty of Chemistry, Wrocław University of Technology, Wybrzeże Wyspiańskiego 27, 50-370 Wrocław, Poland.
Bioelectrochemistry. 2016 Aug;110:19-31. doi: 10.1016/j.bioelechem.2016.02.013. Epub 2016 Feb 27.
Nanocarriers and electroporation (also named electropermeabilization) are convenient methods to increase drug transport. In the current study, we present an effective support of drug delivery into cancer cells, utilizing these methods. We compare the efficiency of each of them and their combination. Multifunctional solid lipid nanoparticles (SLNs) loaded with a cyanine-type IR-780 - acting as a diagnostic agent and a photosensitizer, and a flavonoid derivative - baicalein (BAI) or fisetin (FIS) as a therapeutic cargo - were fabricated via solvent-diffusion method. A therapy supplemented with flavonoids may provide a more precise method to apply desirable lower drug doses and is more likely to result in lower toxicity and a decrease in tumor growth. The SLNs were stabilized with Phospholipon 90G at various concentrations; cetyl palmitate (CP) was applied as a solid matrix. The obtained nanosystems were characterized by dynamic light scattering (size along with size distribution), UV-vis (cargos encapsulation efficiency) and atomic force microscopy (morphology and shape). The obtained SLNs were used as drug carriers alone and in combination with electropermeabilization induced by millisecond pulsed electric fields of high intensity. Two cell lines were selected for the study: LoVo and CHO-K1. The viability was assessed after electroporation alone, the use of electroporation and nanoparticles, and nanoparticles or drugs alone. The intracellular accumulation of cyanine IR-780 and the impact on intracellular structure organization of cytoskeleton was visualized with confocal microscopy method with alpha-actin and beta-tubulin. In this study, the efficacy of nanoparticles with mixed cargo, additionally enhanced by electroporation, is demonstrated to act as an anticancer modality to eliminate cancer cells.
纳米载体和电穿孔(也称为电渗透)是增加药物转运的便捷方法。在本研究中,我们利用这些方法为药物递送至癌细胞提供了一种有效的支持。我们比较了它们各自以及它们组合的效率。通过溶剂扩散法制备了负载花青型IR-780(用作诊断剂和光敏剂)以及类黄酮衍生物黄芩素(BAI)或漆黄素(FIS)(用作治疗性药物)的多功能固体脂质纳米粒(SLN)。补充类黄酮的疗法可能提供一种更精确的方法来应用所需的较低药物剂量,并且更有可能降低毒性并减少肿瘤生长。SLN用不同浓度的磷脂90G稳定;十六烷基棕榈酸酯(CP)用作固体基质。通过动态光散射(尺寸及其分布)、紫外可见光谱(药物包封效率)和原子力显微镜(形态和形状)对所得纳米系统进行表征。所得的SLN单独用作药物载体,并与高强度毫秒脉冲电场诱导的电渗透联合使用。选择了两种细胞系进行研究:LoVo和CHO-K1。在单独进行电穿孔、使用电穿孔和纳米颗粒以及单独使用纳米颗粒或药物后评估细胞活力。用共聚焦显微镜方法结合α-肌动蛋白和β-微管蛋白观察花青IR-780的细胞内积累以及对细胞骨架细胞内结构组织的影响。在本研究中,具有混合药物的纳米颗粒的功效,通过电穿孔进一步增强,被证明可作为一种抗癌方式来消除癌细胞。