Cai, Minming (2021) Characterization of Chitosan Nanocomplexes for the Delivery of Double Stranded MicroRNA to Glioblastoma Cells. MSc by research thesis, University of Leeds.
Abstract
Nanoscale drug delivery systems has gained significant traction, particularly for rare diseases still lacking a curative treatment such as glioblastoma. To complete the treating system of glioblastoma employing chitosan (CS) as a potential non-viral gene delivery vehicle, the present study aimed to assess the non-viral delivery of microRNA (miRNA) to glioblastoma cells (U251 cell line) and hence, its potential to inhibit the intracellular synthesis of proteins, while exerting a cytotoxic effect in this cell line. To this end a research, chitosan (molecular weight (Mw): 25 Kg/mol, degree of acetylation (DA): 29%) was used to form polyelectrolyte complexes with double-stranded miRNA (ds-miRNA) of [-NH3+]/[-PO4-] molar charge ratio (N/P = 20), and their transfection efficiency was assessed in vitro. The particle size distribution, ζ potential, stability and electrophoretic mobility of CS-miRNA complexes of varying N/P ratios were characterized. The transfection efficiency was evaluated from the cytotoxicity U251 cells using the MTT assay. A linear dose-response dependence was established and the cell viability upon treating the cells with CS-ds-miRNA complexes (200μM) for 72 hours was reduced up to 55.35%, thus indicating a targeted cytotoxic effect. Neither ds-miRNA nor chitosan applied in free form exerted a cytotoxic effect. The results demonstrate that chitosan (DA of 29%) has the ability to serve as a non-viral gene delivery vector for miRNA in glioblastoma cells. The in vitro proof of concept gained may serve as the basis for future animal and pre-clinical studies, in the roadmap for potential translation to clinical use in cancer therapy.
Metadata
Supervisors: | Goycoolea, Francisco M. and Guo, Yuan |
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Awarding institution: | University of Leeds |
Academic Units: | The University of Leeds > Faculty of Environment (Leeds) The University of Leeds > Faculty of Maths and Physical Sciences (Leeds) > Food Science (Leeds) |
Academic unit: | School of Food Science and Nutrition |
Depositing User: | Mr. MINMING CAI |
Date Deposited: | 15 Jul 2021 10:02 |
Last Modified: | 01 Jul 2022 00:28 |
Open Archives Initiative ID (OAI ID): | oai:etheses.whiterose.ac.uk:28862 |
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