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Contributor: Ryan Williams, PhD., Assistant Professor, Biomedical Engineering, Grove School of Engineering, The City College of New York

Document Type

Thesis

Abstract

Mesoscale nanoparticles (MNPs) for the therapeutic relief of renal ailments can be modulated to deliver various payloads – including biomolecules such as mRNA. In preparing mRNA loaded-MNPs, we observed a point of saturation for mRNA loading of these particles, when aiming to increase the payload per particle. Here, we aimed to circumvent this limitation by incorporating various excipients that interact with mRNA for increased loading. These interactions involved the reduction of mRNA electrostatic repulsion and improving mRNA stability during formulation and release. Thereafter, we tested the encapsulation efficiency of these modified particles and compared it to our original formulation. Further, we performed cytotoxicity screens and executed functionality tests. These functionality tests included studies of pharmacokinetics, mRNA uptake studies in vitro using qPCR and protein expression through fluorescence microscopy and flow cytometry. Quality assurance to ensure that the particles maintained their mesoscale size range, necessary for kidney targeting, was also an important parameter in this study. Ultimately, we observed that our formulations modified with 1,2-dioleoyl-3-trimethylammonium-propane , trehalose or calcium acetate satisfied these requirements.

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