Masters Theses
Keywords and Phrases
cancer; ionizable lipid nanoparticles; LNP; mRNA delivery; transfection
Abstract
"This thesis investigates the preparation, characterization, and transfection efficiency of various ionizable lipid nanoparticles (LNPs) formulated for the delivery of mRNA encoding enhanced green fluorescent protein (EGFP) in RAW 264.7 macrophages and MDA-MB-231 breast cancer cell lines. The ionizable cationic lipid studied are ALC-035, C12-200, C14-4, PPZ-A10, DLin-MC3-DMA. The study addresses the need for effective gene delivery systems to provide a safe and versatile platform that protects and transports nucleic acids into target cells. LNPs were synthesized using microfluidic mixing methods, incorporating lipid components such as the cationic lipids, DSPE, DOPE, cholesterol and PEG lipids to achieve controlled particle sizes, encapsulation efficiencies, and stability profiles. Dynamic light scattering (DLS) characterized the LNPs immediately post-synthesis and after a seven-day storage period. Particle sizes ranged from 185.7 nm to 247.0 nm initially, with polydispersity index (PI) values between 0.19 and 0.31, indicating stable and uniform formulations. Over a seven-day period, average particle sizes showed less than 10% size increases. Zeta potential values remained close to neutral, between -1.238 mV and -1.786 mV, supporting sustained stability. The formulation was encapsulated across all formulations as confirmed by gel electrophoresis. Transfection efficiency was evaluated using both fluorescence microscopy and flow cytometry. Both cell lines displayed significant expression 24 hours post-transfection with PPZ and C14. Flow cytometry analysis showed that PPZ, C14 formulations led to approximately 70% EGFP-positive cells in RAW 264.7 and 65% in MDA-MB-231, while. These results demonstrate the potential of LNPs as a robust mRNA delivery system, providing valuable insights into nanocarrier optimization for a variety of therapeutic applications especially in cancers"-- Abstract, p. iii
Advisor(s)
Yang, Hu
Committee Member(s)
Elazab, Hany A.
Wang, Jee-Ching
Department(s)
Chemical and Biochemical Engineering
Degree Name
M.S. in Chemical Engineering
Publisher
Missouri University of Science and Technology
Publication Date
Spring 2025
Pagination
x, 39 pages
Note about bibliography
Includes_bibliographical_references_(pages 36-38)
Rights
©2024 Lavanya Bhargava , All Rights Reserved
Document Type
Thesis - Open Access
File Type
text
Language
English
Thesis Number
T 12453
Recommended Citation
Bhargava, Lavanya, "Transfection Of Ionizable Lipid Nanoparticles On Raw 264.7 And Mda-Mb-231" (2025). Masters Theses. 8236.
https://scholarsmine.mst.edu/masters_theses/8236
