Publication Date

12-2017

Date of Final Oral Examination (Defense)

12-8-2017

Type of Culminating Activity

Dissertation

Degree Title

Doctor of Philosophy in Biomolecular Sciences

Department

Biology

Supervisory Committee Chair

Denise G. Wingett, Ph.D.

Supervisory Committee Member

Daniel Fologea, Ph.D.

Supervisory Committee Member

Matthew L. Ferguson, Ph.D.

Supervisory Committee Member

Dmitri Tenne, Ph.D.

Abstract

Nanotechnology has grown exponentially since its inception in the early 1970’s. Since then, bionanotechnological devices and treatment options have significantly improved disease treatments and patient outcomes; however, this rapid growth in consumer related products has also prompted concern. Zinc oxide nanoparticles (nZnO), known for their inherent toxicity and prevalent global use in consumer products and medical applications, have received much of this attention. Significant research efforts have focused on both toxicity remediation through material property modification and the exploitation of these same factors to create potential cancer therapeutics. There is general agreement that the physicochemical properties of nZnO strongly contribute to NP-induced toxicity; however, inconsistencies in the material property characterization methods employed, and an understanding of how those properties influence cytotoxicity in mammalian cells has led to discrepancies in the literature. Additionally, more research is needed to connect the material properties of nZnO to downstream cellular responses. Here, a panel of variably synthesized nZnO was utilized to thoroughly investigate the material properties of the particles as they relate to cytotoxicity, oxidative stress, and transcriptome changes in different mammalian cell types. The goals of this study are three-fold: i) reduce NP agglomeration and sedimentation tendencies within complex media and achieve dispersion stability, ii) define which material property interactions have the greatest potential to affect cellular toxicity, and to iii) examine the preferential toxicity of nZnO towards Jurkat leukemic cells through genetic expression studies.

Chapter 2 highlights the importance of dispersion stability and the effect of fetal bovine serum (FBS) proteins on the dispersion stability, dosimetry and NP-induced cytotoxicity of nZnO in suspension and adherent in vitro cell culture models. The presence of surface adsorbed proteins from the FBS on the nZnO decreased agglomeration and sedimentation potential. Furthermore, FBS-stabilized nZnO dispersions resulted in toxicity increases in suspension cells when compared to unstable dispersions; however, toxicity was decreased in adherent cell models with stable dispersions. These observations indicate that improved dispersion stability leads to increased NP bioavailability for suspension cells and reduced NP sedimentation onto adherent cell layers resulting in more accurate in vitro toxicity assessments.

In Chapter 3, we utilized an expanded panel of nZnO synthesized through wet chemical and high temperature methods, followed by thorough characterization to examine the importance of material property changes in NP-induced toxicity. We found our diverse set of nZnO displayed significant differences in surface reactivity, dissolution potential and cytotoxicity towards cancerous and primary T cells. Additionally, principal component analysis (PCA) suggested that the synthesis procedure conferred unique material properties, and can be a determinant of cellular cytotoxicity. Furthermore, we showed that attributing NP-induced toxicity to one specific material property is shortsighted and that complex interactions between these properties needs to be considered.

Finally, Chapter 4 introduces future work dedicated to investigating transcriptome changes in cancerous and primary T cells exposed to nZnO. Both cell types demonstrated significant up- and down-regulation of genes in a dose-dependent manner. Many significant differentially expressed genes (SDEGs) corresponded to proteins involved in the sequestration and transport of ionic zinc confirming the importance of nZnO in the cytotoxic response. Additional analysis will focus on the importance of specific SDEGs involved in the regulation of oxidative stress pathways, cellular metabolism, inflammation, T cell activation, and protein misfolding in the NP-induced toxicity mechanism.

DOI

https://doi.org/10.18122/B2242C

Included in

Nanomedicine Commons

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