Turiel-Fernández D, Gutiérrez-Romero L, Corte-Rodriguez M, Bettmer J, Montes-Bayón M
Department of Physical and Analytical Chemistry. Faculty of Chemistry. University of Oviedo. Instituto de Investigación Sanitaria Del Principado de Asturias (ISPA), C/ Julián Clavería 8, 33006, Oviedo, Spain.
Department of Physical and Analytical Chemistry. Faculty of Chemistry. University of Oviedo. Instituto de Investigación Sanitaria Del Principado de Asturias (ISPA), C/ Julián Clavería 8, 33006, Oviedo, Spain.
Anal Chim Acta. 2021 May 15;1159:338356. doi: 10.1016/j.aca.2021.338356. Epub 2021 Mar 19.
Ultrasmall iron oxide nanoparticles (<10 nm) were explored here as nanotransporters of cis-diamminetetrachloroplatinum (IV) (a cisplatin prodrug) in cellular models. The coating of the particles containing reactive carboxylic acid groups enabled the formation of a stable conjugate between the prodrug and the nanoparticles using one pot reaction. The nanoconjugate was characterized by different techniques exhibiting diameters of about 6.6 ± 1.0 nm. The use of a hyphenated strategy based on high performance liquid chromatography (HPLC) coupled to inductively coupled plasma mass spectrometry (ICP-MS) permitted the quantitative evaluation of Fe and Pt in the nanoconjugate. Furthermore, the cellular uptake of the synthetic nanoconjugate was explored by single cell-ICP-MS (SC-ICP) which was used for the first time in this type of studies. The experiments in A2780 and A2780cis, sensitive and resistant ovarian cancer cell models respectively, revealed intracellular platinum concentrations of 12 fg/cell and 4 fg/cell, respectively which were 4-fold higher with respect to the uptake of cisplatin in both models. Intracellular drug release from the nanoconjugate was proved by measuring DNA platination in the same cells. In this case, levels of about 250 ng Pt/mg DNA were observed, about 5-fold higher when the nanoconjugate was used in comparison to cisplatin. Furthermore, the differences between the two lines turned to be significantly smaller than in the case of using cisplatin. The quantitative analytical tools developed here provided essential information required to fully characterize the developed nanoplatforms particularly important to overcome drug resistance.
在此研究中，超小氧化铁纳米颗粒（<10纳米）被用作顺二氯二氨铂（IV）（一种顺铂前药）在细胞模型中的纳米转运体。含有反应性羧酸基团的颗粒涂层能够通过一锅法反应在纳米颗粒与前药之间形成稳定的共轭物。该纳米共轭物通过不同技术进行表征，其直径约为6.6±1.0纳米。基于高效液相色谱（HPLC）与电感耦合等离子体质谱（ICP-MS）联用的联用策略允许对纳米共轭物中的铁和铂进行定量评估。此外，通过单细胞ICP-MS（SC-ICP）探索了合成纳米共轭物的细胞摄取情况，这是首次在这类研究中使用。分别在敏感和耐药的卵巢癌细胞模型A2780和A2780cis中进行的实验表明，细胞内铂浓度分别为12 fg/细胞和4 fg/细胞，在这两种模型中，相对于顺铂的摄取量均高出4倍。通过测量同一细胞中的DNA铂化证明了纳米共轭物在细胞内的药物释放。在这种情况下，观察到约250 ng Pt/mg DNA的水平，与使用顺铂相比，使用纳米共轭物时高出约5倍。此外，两条细胞系之间的差异比使用顺铂时明显更小。此处开发的定量分析工具提供了全面表征所开发纳米平台所需的重要信息，这对于克服耐药性尤为重要。
