In Vitro, Non-Invasive Imaging and Detection of Single Living Mammalian Cells Interacting with Bio-Nano-Interfaces
Date of Award:
Doctor of Philosophy (PhD)
S. Clay Isom
Charles D. Miller
Study of bio-nano-interfaces of living mammalian cells will help the identification of cellular alterations (e.g. nucleic acids, amino acids, biomechanics, etc.) due to external stimuli, the design of biomaterials (e.g. nanoparticles, nanotubes) and the investigation on the interaction between cells and bio-nano-interfaces (e.g. cell differentiation on 3D nanostructured materials). The chemical, physical, and mechanical changes of mammalian cells interacting with biomaterials can be investigated by analytical techniques. In this dissertation, cellular responses and property changes are evaluated by non-invasive spectroscopic technique, Raman microscopy (RM), and atomic force microscopy (AFM) combined with traditional biological methods. This dissertation includes the biophysical, biochemical and cytotoxic measurement of cells interacting with bio-nano-interfaces, and this work can be divided into three topics: biomechanics/cellular biopolymers measurement, bio-interfaces and nano-interfaces studies. Cellular biomechanical, biochemical and genetic changes were detected in the topic of biomechanics/cellular biopolymers measurement, and cellular differentiation can be identified by cellular biophysical and biochemical properties. For the bio-interfaces, cellular biochemical and biomechanical properties were affected by BRMS1 expression (a metastasis suppressor) through the study of five metastatic and non-metastatic cancer cells. However, both metastatic and non-metastatic cells exhibited similar patterns of reactive oxygen species (ROS), apoptosis expression and cell viability changes over doxorubicin (DOX) incubation time. Then A549 cells were incubated with diesel exhaust particles (DEP) and an antioxidant resveratrol (RES) to study the effect from DEP and RES, and it was found that RES can decrease DEP-induced destructive effect on cellular structure and enhance DEP-induced biomechanical and inflammatory changes. For the nano-interfaces, hybrid nanoparticles were first developed with the function of fluorescence imaging, Surface-enhanced Raman spectroscopy (SERS) detection and photothermal therapy (PTT). These nanoparticles were applied for single living cells analysis of epidermal growth factor receptor (EGFR) distribution and cancer therapy with high EGFR expression. Additionally, silica coated nanoparticles conjugated with anti-human epidermal growth factor receptor 2 (HER2) were synthesized to increase surface area, light-heat conversion efficiency and biocompatibility. Cancer cells with high-HER2 expression were killed upon an 808 nm laser irradiation. Moreover, trophoblast-derived stem-like cells were cultured on three-dimensional TiO2 nanotubes with Au to study cell viability, morphology and biochemical changes.
Li, Qifei, "In Vitro, Non-Invasive Imaging and Detection of Single Living Mammalian Cells Interacting with Bio-Nano-Interfaces" (2015). All Graduate Theses and Dissertations. 4520.
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