title: Energy independent uptake and release of polystyrene nanoparticles in primary mammalian cell cultures.
abstract: 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.
authors.affiliation: Dipartimento di Biologia, Università di Napoli "Federico II", Complesso Universitario di Monte S Angelo,Via Cinthia 80126 Napoli, Italy.
authors.forename: Ilaria
authors.initials: I
authors.lastname: Fiorentino
authors.affiliation: Dipartimento di Biologia, Università di Napoli "Federico II", Complesso Universitario di Monte S Angelo,Via Cinthia 80126 Napoli, Italy.
authors.forename: Roberto
authors.initials: R
authors.lastname: Gualtieri
authors.affiliation: Dipartimento di Biologia, Università di Napoli "Federico II", Complesso Universitario di Monte S Angelo,Via Cinthia 80126 Napoli, Italy.
authors.forename: Vincenza
authors.initials: V
authors.lastname: Barbato
authors.affiliation: Italian Institute of Technology@CRIB Center for Advanced Biomaterials for Health Care, Largo Barsanti e Matteucci, 53, 80125 Napoli, Italy.
authors.forename: Valentina
authors.initials: V
authors.lastname: Mollo
authors.affiliation: Dipartimento di Biologia, Università di Napoli "Federico II", Complesso Universitario di Monte S Angelo,Via Cinthia 80126 Napoli, Italy.
authors.forename: Sabrina
authors.initials: S
authors.lastname: Braun
authors.affiliation: Dipartimento di Biologia, Università di Napoli "Federico II", Complesso Universitario di Monte S Angelo,Via Cinthia 80126 Napoli, Italy.
authors.forename: Alberto
authors.initials: A
authors.lastname: Angrisani
authors.affiliation: Dipartimento di Biologia, Università di Napoli "Federico II", Complesso Universitario di Monte S Angelo,Via Cinthia 80126 Napoli, Italy.
authors.forename: Mimmo
authors.initials: M
authors.lastname: Turano
authors.affiliation: Dipartimento di Biologia, Università di Napoli "Federico II", Complesso Universitario di Monte S Angelo,Via Cinthia 80126 Napoli, Italy.
authors.forename: Maria
authors.initials: M
authors.lastname: Furia
authors.affiliation: Dipartimento Ingegneria Chimica, dei Materiali e della Produzione Industriale -Piazzale Tecchio, 80126 Napoli, Italy.
authors.forename: Paolo A
authors.initials: PA
authors.lastname: Netti
authors.affiliation: Dipartimento Ingegneria Chimica, dei Materiali e della Produzione Industriale -Piazzale Tecchio, 80126 Napoli, Italy.
authors.forename: Daniela
authors.initials: D
authors.lastname: Guarnieri
authors.affiliation: Dipartimento Ingegneria Chimica, dei Materiali e della Produzione Industriale -Piazzale Tecchio, 80126 Napoli, Italy.
authors.forename: Sabato
authors.initials: S
authors.lastname: Fusco
authors.affiliation: Dipartimento di Biologia, Università di Napoli "Federico II", Complesso Universitario di Monte S Angelo,Via Cinthia 80126 Napoli, Italy. Electronic address: riccardo.talevi@unina.it.
authors.forename: Riccardo
authors.initials: R
authors.lastname: Talevi
mesh_terms: D000584:Amiloride / Q000031:analogs & derivatives / Q000494:pharmacology; D000818:Animals; D001692:Biological Transport; D019086:Bridged Bicyclo Compounds, Heterocyclic / Q000494:pharmacology; D002417:Cattle; D051242:Caveolin 1 / Q000037:antagonists & inhibitors / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism; D002478:Cells, Cultured; D003106:Colon / Q000166:cytology / Q000378:metabolism*; D016503:Drug Delivery Systems; D004705:Endocytosis; D004847:Epithelial Cells / Q000378:metabolism; D005260:Female; D005347:Fibroblasts / Q000378:metabolism; D006801:Humans; D006835:Hydrazones / Q000494:pharmacology; D018613:Microscopy, Confocal; D053758:Nanoparticles / Q000378:metabolism*; D009369:Neoplasms; D010057:Oviducts / Q000166:cytology / Q000378:metabolism*; D010316:Particle Size; D011137:Polystyrenes / Q000378:metabolism*; D061251:Primary Cell Culture; D013050:Spectrometry, Fluorescence; D013395:Sucrose / Q000494:pharmacology; D053778:Thiazolidines / Q000494:pharmacology
chemical_list: D019086:Bridged Bicyclo Compounds, Heterocyclic; D051242:Caveolin 1; D006835:Hydrazones; C511794:N'-(3,4-dihydroxybenzylidene)-3-hydroxy-2-naphthahydrazide; D011137:Polystyrenes; D053778:Thiazolidines; D013395:Sucrose; D000584:Amiloride; C037067:latrunculin A; C039614:ethylisopropylamiloride
keywords: Endocytosis/exocytosis; Nanomedicine; Polystyrene nanoparticles; Primary cell culture