Drug Transport and Delivery Research Group, Department of Pharmacy, Faculty of Health Sciences, University of Tromsø The Arctic University of Norway, N-9037 Tromsø, Norway.
Women's Health and Perinatology Research Group, Department of Clinical Medicine, Faculty of Health Sciences, University of Tromsø The Arctic University of Norway, N-9037 Tromsø, Norway.
Int J Mol Sci. 2020 Jul 9;21(14):4847. doi: 10.3390/ijms21144847.
The rather limited success of translation from basic research to clinical application has been highlighted as a major issue in the nanomedicine field. To identify the factors influencing the applicability of nanosystems as drug carriers and potential nanomedicine, we focused on following their fate through fluorescence-based assays, namely flow cytometry and imaging. These methods are often used to follow the nanocarrier internalization and targeting; however, the validity of the obtained results strictly depends on how much the nanosystem's fate can be inferred from the fate of fluorescent dyes. To evaluate the parameters that affect the physicochemical and biological stability of the labeled nanosystems, we studied the versatility of two lipid dyes, TopFluor-PC and Cy5-DSPE, in conventional liposomes utilizing well-defined in vitro assays. Our results suggest that the dye can affect the major characteristics of the system, such as vesicle size and zeta-potential. However, a nanocarrier can also affect the dye properties. Medium, temperature, time, fluorophore localization and its concentration, as well as their interplay, affect the outcome of tracing experiments. Therefore, an in-depth characterization of the labeled nanosystem should be fundamental to understand the conditions that validate the results within the screening process in optimization of nanocarrier.
从基础研究到临床应用的转化相当有限，这已成为纳米医学领域的一个主要问题。为了确定影响纳米系统作为药物载体和潜在纳米药物适用性的因素，我们专注于通过荧光检测法（即流式细胞术和成像）来跟踪纳米系统的命运。这些方法常用于跟踪纳米载体的内化和靶向性；然而，获得的结果的有效性严格取决于可以从荧光染料的命运推断出纳米系统的命运有多少。为了评估影响标记纳米系统理化和生物学稳定性的参数，我们使用明确的体外检测方法研究了两种脂质染料，即 TopFluor-PC 和 Cy5-DSPE，在常规脂质体中的多功能性。我们的结果表明，染料可以影响系统的主要特征，如囊泡大小和 zeta 电位。然而，纳米载体也可以影响染料的性质。介质、温度、时间、荧光团定位及其浓度，以及它们之间的相互作用，都会影响跟踪实验的结果。因此，对标记纳米系统进行深入的表征应该是理解筛选过程中验证结果的条件以及优化纳米载体的基础。
