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  • Efficiency of Chitosan/Hyaluronan-Based mRNA Delivery Systems In Vitro: Influence of Composition and Structure.

Efficiency of Chitosan/Hyaluronan-Based mRNA Delivery Systems In Vitro: Influence of Composition and Structure.

Journal of pharmaceutical sciences (2020-01-01)
Ousamah Younoss Soliman, Mohamad Gabriel Alameh, Gregory De Cresenzo, Michael D Buschmann, Marc Lavertu
RESUMO

Messenger RNA (mRNA)-containing nanoparticles were produced by electrostatic complexation with a library of pharmaceutical grade chitosans with different degrees of deacetylation and hyaluronic acids (HAs) (native vs. sulfated). Polymer length (Mn), HA degree of sulfation (DS), and amine to phosphate to carboxyl + sulfate (from HA) ratio (N:P:C) were controlled. In vitro transfections were performed in the presence/absence of trehalose and at different pH. Particle size and ζ-potential were correlated with transfection efficiency. Polymer length and charge densities (degree of deacetylation, degree of sulfation) of both HA and chitosan had a direct influence on transfection efficiency through modulation of avidity to mRNA. N:P:C ratio, trehalose, mixing concentration, and nucleic acid dose influenced transfection efficiency with optimized formulations reaching ∼60%-65% transfection efficiency relative to commercially available lipid control with no apparent toxicity for transfection at slightly acidic pH 6.5.

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Sigma-Aldrich
D-(+)-Trealose, ≥99% (HPLC), from Saccharomyces cerevisiae
Supelco
Ácido clorídrico, volumetric, 0.1 M HCl (0.1N), endotoxin free
Sigma-Aldrich
Hidróxido de sódio, puriss. p.a., ACS reagent, K ≤0.02%, ≥98.0% (T), pellets
Amersham Hybond® P Western blotting membranes, PVDF, pore size 0.45 μm, roll W × L 300 mm × 4 m, pkg of 1 ea