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Cell Uptake and Validation of Novel PECs for Biomedical Applications.

Journal of drug delivery (2011-08-31)
Ilaria E Palamà, Mariarosaria Musarò, Addolorata M L Coluccia, Stefania D'Amone, Giuseppe Gigli
ABSTRACT

This pilot study provides the proof of principle for biomedical application of novel polyelectrolyte complexes (PECs) obtained via electrostatic interactions between dextran sulphate (DXS) and poly(allylamine hydrochloride) (PAH). Scanning electron microscopy (SEM) and atomic force microscopy (AFM) showed that DXS/PAH polyelectrolyte complexes were Monodispersed with regular rounded-shape features and average diameters of 250 nm at 2 : 1 weight ratios of DXS/PAH. Fluorescently labelled DXS and fluorescein-isothiocyanate- (FITC-)conjugate DXS were used to follow cell uptake efficiency of PECs and biodegradability of their enzymatically degradable DXS-layers by using confocal laser scanning microscopy (CLSM). Moreover, quantitative MTT and Trypan Blue assays were employed to validate PECs as feasible and safe nanoscaled carriers at single-cell level without adverse effects on metabolism and viability.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Thiazolyl Blue Tetrazolium Bromide, powder, BioReagent, suitable for cell culture, suitable for insect cell culture, ≥97.5% (HPLC)
Sigma-Aldrich
Thiazolyl Blue Tetrazolium Bromide, 98%
Sigma-Aldrich
Fetal Bovine Serum, Australia origin, USDA approved, sterile-filtered, suitable for cell culture
Sigma-Aldrich
Fetal Bovine Serum, USA origin, suitable for cell culture
Sigma-Aldrich
Fetal Bovine Serum, USA origin, Heat Inactivated, sterile-filtered, suitable for cell culture
Sigma-Aldrich
Poly(allylamine hydrochloride), average Mw ~17,500 (GPC vs. PEG std.)
Sigma-Aldrich
Poly(allylamine hydrochloride), Mw 40,000-150,000