Graduation Year

2009

Document Type

Dissertation

Degree

Ph.D.

Degree Granting Department

Chemical and Biomedical Engineering

Major Professor

Michael VanAuker, Ph.D.

Co-Major Professor

Mark Jaroszeski, Ph.D.

Committee Member

Joel Strom, M.D.

Committee Member

Nathan Gallant, Ph.D.

Committee Member

Ryan Toomey, Ph.D.

Keywords

targeted drug delivery, polyethylene glycol, nanoparticle, liposome, inflammation

Abstract

The research presented in this dissertation describes the creation and characterization of a novel antibody-vesicle conjugate modified with polyethylene glycol (PEG) that possesses enhanced binding to and uptake by inflammation-activated endothelial cells with improved storage stability and longer shelf-life and potential reduction in immunogenic potential compared to previous designs.

Targeted drug delivery provides an effective means of delivering therapeutic concentrations of a drug to the site or organ of action. The drug is delivered using a niosome, a vesicle with an aqueous core and a bilayer membrane composed of non-ionic surfactants and cholesterol. Antibodies that recognize specific cell antigens are attached to the niosome to complete the targeting molecule, an immuno-niosome (IN). When functionalized PEG, a water soluble, biologically inert polymer, is attached to proteins, it can protect the protein, increasing its half-life in vivo. The immuno-niosome synthesis process is modified to include PEG incorporation into the niosome membrane, a process known as PEGylation.

Since the vasculature connects the entire body, immuno-niosomes targeted to endothelial cells were used. When endothelial cells are activated during disease, stress and injury, certain receptors are expressed and upregulated. One such receptor, CD44, is upregulated in response to vascular inflammation associated with atherosclerosis.

The research hypothesis is that the addition of polyethylene glycol to the drug delivery vesicle (immuno-niosome) using cyanuric chloride linking chemistry will improve colloidal stability and binding performance of the PEGylated immuno-niosomes to endothelial cell surface receptors expressed during an inflammatory response. The research presented in this dissertation provides the following evidence to support this hypothesis:

Construction and characterization of a modified drug delivery vesicle using non-ionic surfactants conjugated with PEG and functionalized with antibodies against endothelial cell surface receptors (PEGylated immuno-niosomes) improves the colloidal stability over previously designed vesicles.

Binding of PEGylated CD44-IN to activated endothelial cells is improved over previously designed vesicles.

Binding of PEGylated CD44-IN to activated endothelial cells under physiological conditions (flow) is demonstrated.

Uptake of PEGylated immuno-niosomes by activated or injured endothelial cells is demonstrated using confocal microscopy.

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