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University of Sydney

1. Khanal, Dipesh. Toxicological considerations of nanoparticles using two and three dimensional liver models: Holistic approach to nanoparticle characterization and nanotoxicity assessment .

Degree: 2018, University of Sydney

Nanoparticles became ‘indispensable’ components of cosmetics, food products, biosensor and therapeutics. Daily consumption of nanoparticles exceeds 1 mg/kg of body mass for adults and more importantly 2 mg/kg of body mass for kids (few trillions of nanoparticles per day). There is increasing evidence that nanoparticles contribute to the loss of cell function and may be responsible for increase in auto immune disease, asthma and autism. The impact of nanomaterials on health still remains unexplored. Conventional nanosafety assessment tools have limited capability and resolution, these methods are incapable to provide robust evidence of the nano-biointeractions and the toxicity associated to these engineered nanoparticles precisely. In the current study, we developed a holistic approach to characterize the safety of nanoparticles which had the advantage of high-throughput screening abilities as well as sub nano level resolution. We confirmed the dynamic process of protein corona formation on the nanodiamond particles (NDs) with the tools as nanoIsothermal calorimetry (nanoITC), matrix assisted laser desorption/ionization time of flight spectrometry (MALDI-TOF). In addition, we used nanoscale infrared spectroscopy (AFM-IR) to detect interactions of single nanoparticle with proteins within cells. We demonstrated that NDs induced alterations in the secondary protein structure evidenced by the formation of antiparallel β-sheet and β-turns. For the first time Lorentz contact resonance spectroscopic technique (LCR) was used to investigate the uptake and localization of nanoparticles based on local viscoelasticity. Nanomechanical studies using molecular force probe (MFP) for cells treated with NDs showed concentration-dependent change in the cell stiffness. Since stiffness represents cell functionality these results imply that NDs induce some toxic effects. All the toxicity studies of NDs were conducted using 2D and 3D liver models (High-throughput screening tool). Monolayer cell culture model (Fao cells) and scaffold-free tissue model generated using magnetic levitation were used as 2D and 3D models. Nanotoxicity was assessed using:3D holographic tomographic microscopy and serial block face scanning electron microscopy (uptake and distribution), live cell imaging (cell morphology and migration), viability (live/dead), vitality (CCK-8), tight junction integrity (impedance measurement), 3D ring closure and spheroid dot assay. These in vitro assays showed that NDs induced concentration-dependent toxicity in both 2D & 3D models. In summary, our results highlight that the tools such as 3D ring closure and dot assay along with, nanoITC, MALDI-TOF, LCR, AFM-IR, MFP, not only provide fundamental understanding of interactions of material and biological milieu at nanolevel but also enable precise assessment of nanotoxicity.

Subjects/Keywords: Nanosafety; nanodiamond; AFM; biospectroscopy; Liver model; Nanocharacterization

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6th Edition):

Khanal, D. (2018). Toxicological considerations of nanoparticles using two and three dimensional liver models: Holistic approach to nanoparticle characterization and nanotoxicity assessment . (Thesis). University of Sydney. Retrieved from http://hdl.handle.net/2123/18637

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16th Edition):

Khanal, Dipesh. “Toxicological considerations of nanoparticles using two and three dimensional liver models: Holistic approach to nanoparticle characterization and nanotoxicity assessment .” 2018. Thesis, University of Sydney. Accessed September 19, 2019. http://hdl.handle.net/2123/18637.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7th Edition):

Khanal, Dipesh. “Toxicological considerations of nanoparticles using two and three dimensional liver models: Holistic approach to nanoparticle characterization and nanotoxicity assessment .” 2018. Web. 19 Sep 2019.

Vancouver:

Khanal D. Toxicological considerations of nanoparticles using two and three dimensional liver models: Holistic approach to nanoparticle characterization and nanotoxicity assessment . [Internet] [Thesis]. University of Sydney; 2018. [cited 2019 Sep 19]. Available from: http://hdl.handle.net/2123/18637.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Khanal D. Toxicological considerations of nanoparticles using two and three dimensional liver models: Holistic approach to nanoparticle characterization and nanotoxicity assessment . [Thesis]. University of Sydney; 2018. Available from: http://hdl.handle.net/2123/18637

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

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