Intracellular trafficking and cellular uptake mechanism of PHBV nanoparticles for targeted delivery in epithelial cell lines

Abstract

BackgroundNanotechnology is a science that involves imaging, measurement, modeling and a manipulation of matter at the nanometric scale. One application of this technology is drug delivery systems based on nanoparticles obtained from natural or synthetic sources. An example of these systems is synthetized from poly(3-hydroxybutyrate-co-3-hydroxyvalerate), which is a biodegradable, biocompatible and a low production cost polymer. The aim of this work was to investigate the uptake mechanism of PHBV nanoparticles in two different epithelial cell lines (HeLa and SKOV-3).ResultsAs a first step, we characterized size, shape and surface charge of nanoparticles using dynamic light scattering and transmission electron microscopy. Intracellular incorporation was evaluated through flow cytometry and fluorescence microscopy using intracellular markers. We concluded that cellular uptake mechanism is carried out in a time, concentration and energy dependent way. Our results showed that nanoparticle uptake displays a cell-specific pattern, since we have observed different colocalization in two different cell lines. In HeLa (Cervical cancer cells) this process may occur via classical endocytosis pathway and some internalization via caveolin-dependent was also observed, whereas in SKOV-3 (Ovarian cancer cells) these patterns were not observed. Rearrangement of actin filaments showed differential nanoparticle internalization patterns for HeLa and SKOV-3. Additionally, final fate of nanoparticles was also determined, showing that in both cell lines, nanoparticles ended up in lysosomes but at different times, where they are finally degraded, thereby releasing their contents.ConclusionsOur results, provide novel insight about PHBV nanoparticles internalization suggesting that for develop a proper drug delivery system is critical understand the uptake mechanism.Electronic supplementary materialThe online version of this article (doi:10.1186/s12951-016-0241-6) contains supplementary material, which is available to authorized users.

Interaction of γ-Fe2O3 nanoparticles with Citrus maxima leaves and the corresponding physiological effects via foliar application

“…Nowadays, nanomaterials become a hotspot of research interests and attract the attention of many researchers. A variety of nanoparticles (NPs) have been studied on human cells [4, 5], animal cells [6] and plants [7] about their toxicity or applications. As one of the most widely explored and applied nanomaterials, iron oxide nanoparticles (γ-Fe 2 O 3 NPs) are widely used in medical diagnostics, controlled drug release, separation technologies and environmental engineering [8].…”

Bilirubin detoxification using different phytomaterials: characterization and in vitro studies

“…It has a large size distribution of 170-700 nm, with average size of 344 nm. 18,40 Figure 3A shows that most particles were ,200 nm, with an average size of 135 nm. Figure 3B shows a narrow size distribution, with an average size of 284 nm.…”

“…Figure 3C shows a large size distribution of 170-700 nm, with average size of 344 nm. 18,40 Figure 4 shows ζ-potential values of -10.1 mV for date-pit nano-AC, -26.3 mV for jojoba-seed nano-AC, and -42.7 mV for microalgae nano-AC in H 2 O suspension. Datepit nano-AC had small negative charge on the surface, which indicates less stability and affinity to agglomerate with time into clusters, whereas the microalgae nano-AC produced had high stability and homogeneity, due to the repulsive force among particles.…”

“…28,37 Cells were washed and permeabilized with 0.1% Triton X-100 for 5 minutes, fluorescein isothiocyanate (FITC)-labeled phalloidin added, and kept incubated for 40 minutes. [38][39][40] Cells were washed with DPBS and 0.1% Triton X-100 for 4 minutes. Samples were mounted with DAPI-Fluoroshield (Sigma-Aldrich) for 10 minutes, then washed to remove the excess DAPI.…”

“…Samples were mounted with DAPI-Fluoroshield (Sigma-Aldrich) for 10 minutes, then washed to remove the excess DAPI. 40 Finally, coverslips were fixed on frosted slides with the DPX mountant. Similarly, the 3D cultured cells were allowed to grow in Matrigel and collagen for 10 days.…”