Merck

Energy independent uptake and release of polystyrene nanoparticles in primary mammalian cell cultures.

Experimental cell research (2014-09-24)
Ilaria Fiorentino, Roberto Gualtieri, Vincenza Barbato, Valentina Mollo, Sabrina Braun, Alberto Angrisani, Mimmo Turano, Maria Furia, Paolo A Netti, Daniela Guarnieri, Sabato Fusco, Riccardo Talevi
RESUMEN

Nanoparticle (NPs) delivery systems in vivo promises to overcome many obstacles associated with the administration of drugs, vaccines, plasmid DNA and RNA materials, making the study of their cellular uptake a central issue in nanomedicine. The uptake of NPs may be influenced by the cell culture stage and the NPs physical-chemical properties. So far, controversial data on NPs uptake have been derived owing to the heterogeneity of NPs and the general use of immortalized cancer cell lines that often behave differently from each other and from primary mammalian cell cultures. Main aims of the present study were to investigate the uptake, endocytosis pathways, intracellular fate and release of well standardized model particles, i.e. fluorescent 44 nm polystyrene NPs (PS-NPs), on two primary mammalian cell cultures, i.e. bovine oviductal epithelial cells (BOEC) and human colon fibroblasts (HCF) by confocal microscopy and spectrofluorimetric analysis. Different drugs and conditions that inhibit specific internalization routes were used to understand the mechanisms that mediate PS-NP uptake. Our data showed that PS-NPs are rapidly internalized by both cell types 1) with similar saturation kinetics; 2) through ATP-independent processes, and 3) quickly released in the culture medium. Our results suggest that PS-NPs are able to rapidly cross the cell membrane through passive translocation during both uptake and release, and emphasize the need to carefully design NPs for drug delivery, to ensure their selective uptake and to optimize their retainment in the targeted cells.

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Trypan Blue solution, 0.4%, liquid, sterile-filtered, suitable for cell culture
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5-(N-Ethyl-N-isopropyl)amiloride
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Trypan Blue solution, 0.4%, for microscopy
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