Investigating the impact of metallic nanoparticles on biological systems and lysosomal function

Cook, Sophie 2022. Investigating the impact of metallic nanoparticles on biological systems and lysosomal function. PhD Thesis, Cardiff University. Item availability restricted.

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Abstract

Metallic nanoparticles, including iron oxide nanoparticles (IONPs) are emerging as a component of particulate matter air pollution that is being linked to cellular and organism-level toxicity in experimental studies. While IONPs, in particular superparamagnetic iron oxide nanoparticles (SPIONs), such as magnetite, are also being developed as tools for bioscience-based research, and for these kinds of applications it is important to use nanoparticles with low cytotoxicity and high biocompatibility. IONPs can be endocytosed by cells and trafficked through the endolysosomal system to the lysosome. The acidic and degradative environment of the lysosome can promote degradation of IONPs, releasing free iron, which is linked to production of reactive oxygen species (ROS), mainly through Fenton reactions. The production of ROS is believed to be a main driver of IONP toxicity. This thesis aimed to develop cellular and in vivo (zebrafish) models of iron oxide nanoparticle (IONP) treatment and identify affected pathways, phenotypes and behavioural responses, identifying nanoparticle features associated with greater toxicity or greater biocompatible. Investigation of the effects of HEPES, and other cell culture buffers, suggested high concentrations of HEPES buffer and common buffer concentrations of PIPES buffer can affect lysosomal phenotypes in CHO H1 cells. This data being used to select cell culture buffers and concentrations for use in further experiments to prevent buffer-related lysosomal phenotypes. This lab has developed a magnetic lysosomal extraction technique using a dextran coated IONP, labelled LRL, that is able to extract high yields of lysosomes from storage disease cells that are difficult to isolate with other extraction methods due to changes in lysosomal density. Comparison of the LRL nanoparticle, with two similar dextran associated IONPs suggested strong differences in toxicity across a range of cellular pathways and phenotypes between the nanoparticles despite similarity in structure, while validating the biocompatibility of LRL nanoparticles. Therefore, the toxicity of LRL nanoparticles was also investigated in an embryonic zebrafish development model (in vivo), with zebrafish models being increasingly utilised for investigation of lysosomal storage diseases. These experiments determined that this nanoparticle was also highly biocompatible in the developing zebrafish and could also be used to develop a zebrafish magnetic lysosomal extraction protocol.

Item Type:	Thesis (PhD)
Date Type:	Completion
Status:	Unpublished
Schools:	Biosciences
Subjects:	Q Science > Q Science (General)
Date of First Compliant Deposit:	31 May 2022
Date of Acceptance:	31 May 2022
Last Modified:	31 May 2023 01:30
URI:	https://orca.cardiff.ac.uk/id/eprint/150170

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