title: Low molecular weight poly (2-dimethylamino ethylmethacrylate) polymers with controlled positioned fluorescent labeling: Synthesis, characterization and in vitro interaction with human endothelial cells.
abstract: Poly (2-dimethylamino ethylmethacrylate) (PDMAEMA) is an attractive non-degradable polymer studied as nonviral vector for gene delivery but it can be also adopted for delivery of other biopharmaceutical drugs. As a parenteral carrier, the PDMAEMA free form (FF) might interact with tissues and cells. Few data are available on its selective internalization and efflux from cells, while the majority of studies published have followed the distribution of DNA complexed with PDMAEMA. In order to address polycation safety, the first aim was to synthesize by atom transfer radical polymerisation (ATRP) fluorescent labeled PDMAEMA of low molecular weight (Mw) (below 15 kDa), controlling the position and density of fluorescein. The second goal was to analyze the possible difference in uptake and subcellular distribution of this labeled FF polycation between human umbilical vein endothelial cells (HUVEC) and hCMEC/D3 cells. These two cell lines have been chosen in order to detect selectivity towards the blood-brain barrier (BBB). In both cases, polycation was detected along the plasma membrane followed by progressive migration to the peri-nuclear region, where it overlapped with lysosomal structures. The analysis by fluorescence-activated cell sorting (FACS) of the PDMAEMA uptake by hCMEC/D3 cells showed a significant (p<0.05) inhibition (40%) in presence of 2-dexoxy-D-glucose inhibitor, a result supporting an energy-dependence mechanism(s). Cytotoxicity study showed that low Mw PDMAEMA (10 kDa) lead to a minor cytotoxicity compared to the higher ones. As main conclusion this study highlights the similitude in cell trafficking of FF PDMAEMA and data previously reported for PDMAEMA/DNA complexes.
authors.affiliation: Interfacultary Research Center of Biomaterials (CEIB), University of Liège, Institute of Chemistry, Building B6C, Sart-Tilman, Liège (ZIP code 4000), Belgium. Electronic address: luca.flebus@ulg.ac.be.
authors.forename: Luca
authors.initials: L
authors.lastname: Flebus
authors.affiliation: Interfacultary Research Center of Biomaterials (CEIB), University of Liège, Institute of Chemistry, Building B6C, Sart-Tilman, Liège (ZIP code 4000), Belgium. Electronic address: flombart@ulg.ac.be.
authors.forename: François
authors.initials: F
authors.lastname: Lombart
authors.affiliation: Interfacultary Research Center of Biomaterials (CEIB), University of Liège, Institute of Chemistry, Building B6C, Sart-Tilman, Liège (ZIP code 4000), Belgium. Electronic address: csevrin@ulg.ac.be.
authors.forename: Chantal
authors.initials: C
authors.lastname: Sevrin
authors.affiliation: Cardiovascular Research Center, Avenue de l'hôpital 13, Liège (ZIP code 4000), Belgium. Electronic address: jo.defraigne@ulg.ac.be.
authors.forename: Jean-Olivier
authors.initials: JO
authors.lastname: Defraigne
authors.affiliation: Hemostasis and Thrombosis center, CHU, Avenue de l'hôpital 13, Liège (ZIP code 4000), Belgium. Electronic address: Pierre.Peters@chu.ulg.ac.be.
authors.forename: Pierre
authors.initials: P
authors.lastname: Peters
authors.affiliation: Centre for Pharmaceutical Nanotechnology and Nanotoxicology, Department of Pharmacy, Faculty of Health and Medical Sciences and NanoScience Centre, University of Copenhagen, Universitetsparken 2, Copenhagen (ZIP code 2100), Denmark. Electronic address: lpar@nanotech.dtu.dk.
authors.forename: Ladan
authors.initials: L
authors.lastname: Parhamifar
authors.affiliation: Department of Physiology, CARIM, Maastricht University, Universiteitssingel 50, ER Maastricht (ZIP code 6229ER),The Netherlands. Electronic address: d.molin@maastrichtuniversity.nl.
authors.forename: Daniel G M
authors.initials: DGM
authors.lastname: Molin
authors.affiliation: Interfacultary Research Center of Biomaterials (CEIB), University of Liège, Institute of Chemistry, Building B6C, Sart-Tilman, Liège (ZIP code 4000), Belgium. Electronic address: C.Grandfils@ulg.ac.be.
authors.forename: Christian
authors.initials: C
authors.lastname: Grandfils
mesh_terms: D001692:Biological Transport; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D019793:Fluorescein / Q000737:chemistry / Q000494:pharmacology; D005456:Fluorescent Dyes / Q000737:chemistry / Q000494:pharmacology; D061307:Human Umbilical Vein Endothelial Cells / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D008689:Methacrylates / Q000737:chemistry / Q000494:pharmacology; D008970:Molecular Weight; D009757:Nylons / Q000737:chemistry / Q000494:pharmacology
chemical_list: D005456:Fluorescent Dyes; D008689:Methacrylates; D009757:Nylons; C407037:poly(2-(dimethylamino)ethyl methacrylate); D019793:Fluorescein
keywords: Biomedical applications; Cell trafficking; Functional polymers; Poly(2-dimethylamino-ethylmethacrylate); Polycations