Graduation Year

2020

Document Type

Thesis

Degree

M.S.

Degree Name

Master of Science (M.S.)

Degree Granting Department

Medical Sciences

Major Professor

Subhra Mohapatra, Ph.D.

Committee Member

Shyam Mohapatra, Ph.D.

Committee Member

Eleni Markoutsa, Ph.D.

Keywords

Nanoparticles, Transfection, Posterior eye diseases, Non-viral vectors

Abstract

Gene therapy holds promise for treating a wide range of diseases including cancer and several other polygenic and monogenic disorder arising in different organs. Due to its immune privilege nature, eye is an ideal organ for gene therapy,as various visual pathologies arise from gene defect and can lead to partial or complete vision loss. The last decade, several nanoparticles have been developed and tested for their transfection efficiency in vitro and in vivo. Among them, dendrimers with small size, good physicochemical properties and endosomal escape activity have shown high transfection efficiency in various cell lines. In an attempt to further increase the transfection efficiency of the dendrimers, other molecules can be conjugated on the surface of dendrimers to facilitate more efficient cell entry and transfection. Herein, β-cyclodextrin was conjugated on the surface of the PAMAM dendrimer and the effect of this conjugation was tested for transfection efficiency, cellular uptake and toxicity of PAMAM dendrimer in Retinal Epithelium Pigment cells (ARPE). The successful conjugation of cyclodextrin on the PAMAM surface was confirmed by NMR. Different cyclodextrin to PAMAM ratios were also tried in an attempt to optimize the nanoformulation. The cyclodextrin-conjugated dendrimers were then characterized for particle size, zeta potential and morphology. Furthermore, cyclodextrin-conjugated dendrimers were complexed with plasmid and characterized using gel electrophoresis to confirm the successful formation of the complex. In conclusion, the cyclodextrin-PAMAM (CyD-PAMAM) conjugation improves the uptake and transfection efficiency of PAMAM dendrimer without increasing its cell toxicity significantly.

Download

Share

COinS