ORCID Identifier(s)

0000-0003-4824-9502

Graduation Semester and Year

Summer 2025

Document Type

Thesis

Degree Name

Doctor of Philosophy in Biomedical Engineering

Department

Bioengineering

First Advisor

Dr. Kytai T. Nguyen

Second Advisor

Dr. George Alexandrakis

Third Advisor

Dr. Liping Tang

Fourth Advisor

Dr. Hao Xu

Fifth Advisor

Dr. Yaowu Hao

Abstract

Peripheral arterial disease (PAD) is a progressive vascular disorder marked by the narrowing or occlusion of peripheral arteries, leading to chronic ischemia, tissue damage, and in severe cases, limb amputation. Current therapeutic strategies, including pharmacological treatments and surgical revascularization, often fall short—particularly for elderly patients or those with significant comorbidities—underscoring the need for minimally invasive, site-specific alternatives.

We hypothesized that targeted delivery of therapeutic genes using nanoparticles engineered for active endothelial binding could enhance site-specific accumulation in ischemic tissue and improve treatment outcomes in PAD. To test this, we developed poly(lactic-co-glycolic acid) (PLGA) nanoparticles functionalized with anti-ICAM-1 antibodies to actively target inflamed endothelium, where ICAM-1 is significantly upregulated during PAD progression.

Nanoparticles were synthesized via a double emulsion solvent evaporation method and characterized using dynamic light scattering, zeta potential analysis, transmission electron microscopy, and biochemical assays to evaluate size, charge, morphology, and antibody conjugation. To overcome limitations of conventional ensemble-averaged techniques, we further applied emerging single-particle analytical methods—solid-state nanopores, plasmonic nanopores, and nanopipette sensing—to investigate nanoparticle heterogeneity at the individual particle level. These platforms revealed variability in size, charge, and ligand conjugation efficiency, demonstrating their value in quality control and formulation refinement.

In vitro studies using TNF-α–activated endothelial progenitor cells showed that ICAM-1–targeted nanoparticles achieved superior cellular uptake and gene transfection compared to non-targeted controls. In vivo biodistribution in a mouse hindlimb ischemia model confirmed selective accumulation of targeted nanoparticles in ischemic muscle, validating the targeting strategy and highlighting the potential for tissue-specific delivery.

In conclusion, this dissertation presents a multifaceted approach that combines nanoparticle engineering, molecular targeting, and advanced characterization to address current barriers in PAD therapy. The findings support the feasibility and promise of ICAM-1–targeted PLGA nanoparticles as a platform for minimally invasive, precision gene delivery in ischemic vascular disease.

Keywords

nanomedicine, active targeted nanoparticles, peripheral arterial disease

Disciplines

Biological Engineering

License

Creative Commons Attribution 4.0 International License
This work is licensed under a Creative Commons Attribution 4.0 International License.

Available for download on Wednesday, August 11, 2027

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