Abstract
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.
Highlights
Nanomedicine has been proposed as the superior approach within advanced drug therapy, able to respond to the ever growing demands of various diseases
Liposomes made of natural phospholipids offer reassuring safety profiles [4] and become one of the most studied nanomedicines
Considering liposomes as nanocarriers, liposomes are most often labeled by the fluorescent dye lipids incorporated within liposomal bilayers
Summary
Nanomedicine has been proposed as the superior approach within advanced drug therapy, able to respond to the ever growing demands of various diseases. It is important to consider that the findings are based on two assumptions: i) the dye does not alter the interactions of nanocarriers with cells/tissues; ii) the dye encapsulated/incorporated in the nanocarrier is stable and remains associated with the carrier throughout the process. These assumptions are not always correct [6]. The dye dissociations are not detected in the commonly applied in vitro assays used to determine dye leakage For both (i) dye-induced modification of cellular fate and (ii) dye leakage, the dye loses its primary function of tracking the system behavior; as a result, the fate of the dye is followed rather than the fate of the dye-labeled nanocarrier
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