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A New Hyaluronic Acid Derivative Obtained from Atom Transfer Radical Polymerization as a siRNA Vector for CD44 Receptor Tumor Targeting.

A New Hyaluronic Acid Derivative Obtained from Atom Transfer Radical Polymerization as a siRNA Vector for CD44 Receptor Tumor Targeting.

Macromolecular bioscience (2015-07-03)

Fabio Salvatore Palumbo, Antonella Bavuso Volpe, Flavia Bongiovì, Giovanna Pitarresi, Gaetano Giammona

ABSTRACT

Two derivatives of hyaluronic acid (HA) have been synthesized by atom transfer radical polymerization (ATRP), starting from an ethylenediamino HA derivative (HA-EDA) and by using diethylaminoethyl methacrylate (DEAEMA) as a monomer for polymerization. Both samples, indicated as HA-EDA-pDEAEMA a and b, are able to condense siRNA, as determined by gel retardation assay and resulting complexes show a size and a zeta potential value dependent on polymerization number, as determined by dynamic light scattering measurements. In vitro studies performed on HCT 116 cell line, that over express CD44 receptor, demonstrate a receptor mediated uptake of complexes, regardless of their surface charge.

MATERIALS

Product Number

Brand

Product Description

Sigma-Aldrich

Acetone, ACS reagent, ≥99.5%

Sigma-Aldrich

Acetone, puriss., meets analytical specification of Ph. Eur., BP, NF, ≥99% (GC)

Sigma-Aldrich

Acetone, ACS reagent, ≥99.5%

Sigma-Aldrich

Acetone, Laboratory Reagent, ≥99.5%

Sigma-Aldrich

Acetone, histological grade, ≥99.5%

Sigma-Aldrich

Acetone, puriss. p.a., ACS reagent, reag. ISO, reag. Ph. Eur., ≥99.5% (GC)

Sigma-Aldrich

Tetrabutylammonium hydroxide solution, 1.0 M in methanol

Sigma-Aldrich

Tetrabutylammonium hydroxide solution, 53.5-56.5% in H₂O

Sigma-Aldrich

Tetrabutylammonium hydroxide solution, 40 wt. % in H₂O

Sigma-Aldrich

Copper(II) bromide, 99%

Sigma-Aldrich

Copper(II) bromide, 99.999% trace metals basis

Sigma-Aldrich

2-Bromo-2-methylpropionic acid, 98%

Sigma-Aldrich

Diethylamine, purified by redistillation, 99.5%

Sigma-Aldrich

Deuterium oxide, 99.9 atom % D, contains 1 % (w/w) 3-(trimethylsilyl)-1-propanesulfonic acid, sodium salt (DSS)

Sigma-Aldrich

Ethylenediamine, purified by redistillation, ≥99.5%

Sigma-Aldrich

Dimethyl sulfoxide, anhydrous, ≥99.9%

Sigma-Aldrich

8-Octanoyloxypyrene-1,3,6-trisulfonic acid trisodium salt, suitable for fluorescence, ≥90% (HPCE)

Sigma-Aldrich

Deuterium oxide, 99.9 atom % D

Sigma-Aldrich

Diethylamine, puriss. p.a., ≥99.5% (GC)

Sigma-Aldrich

Deuterium oxide, 99.8 atom % D

Sigma-Aldrich

Ethylenediamine, meets USP testing specifications

Sigma-Aldrich

Deuterium oxide, 99.9 atom % D, contains 0.75 wt. % 3-(trimethylsilyl)propionic-2,2,3,3-d₄ acid, sodium salt

Sigma-Aldrich

N-(3-Dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride, ≥99.0% (AT)

Sigma-Aldrich

Dimethyl sulfoxide solution, 50 wt. % in H₂O

Sigma-Aldrich

L-Glutamine

Sigma-Aldrich

Dimethyl sulfoxide, puriss. p.a., ACS reagent, ≥99.9% (GC)

Sigma-Aldrich

Dimethyl sulfoxide, puriss. p.a., dried, ≤0.02% water

Sigma-Aldrich

Ethylenediamine, BioXtra

Sigma-Aldrich

N-(3-Dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride, commercial grade, powder

Sigma-Aldrich

L-Glutamine, γ-irradiated, BioXtra, suitable for cell culture