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Real time monitoring of Cell-Nanoparticles interaction and tracking internalization process by mechanical probing using Atomic Force Microscopy

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Date Issued:
2014
Abstract/Description:
With extensive development of nanotechnology in last few years, scientists have discovered that nanoparticles (NPs) can be used as an efficient Drug Delivery System (DOS). In order to develop better NPs based drug delivery tool, it is imperative to understand the interaction between the NPs and the cell membrane. In our earlier studies, cerium oxide nanoparticles (CNPs) have been reported to have therapeutic properties, specifically against abnormalities associated with oxidative stress. Therefore, CNPs with different sizes and morphology were selected to understand the interaction with cell. We analyzed the mechanical property of human nasal septum tumor cells membranes using Atomic Force Microscopy (AFM) with and without CNPs. In particular, Force-Distance spectroscopy mode was used to estimate the elasticity of cells membrane. Different concentrations (0, 50, 125 and 250 ?M) of CNPs were added to the cells (squamous cells; CCL30) and incubated for different time periods (0, 15, 30 and 60 minutes). Cell membrane elasticity/Young's modulus was calculated using a modified Hertz model. Changes in the cell elasticity were observed in high concentration of CNPs when treated with one hour. Significant changes in cell elasticity were observed at high concentration of CNPs for one hour of incubation. No significant change in cell elasticity was observed over one hour time period for 50 ?M of CNPs. Moreover, by using selected inhibitors to block different cell mediated internalization pathways, we also investigated the correlation between the cellular uptake and the tracking of NPs with their size. Specifically, similar change in cell elasticity was observed after blocking the cell energy production for CNPs with smaller diameter (3-5 nm). On the other hand, bigger size NPs (20-30 nm) showed no change in cell elasticity after blocking the cell energy production. These results indicate that 3-5 nm particles internalize cell by non-energy dependent pathway i.e. passive diffusion whereas 20-30 nm particles entered in cell by energy dependent pathways i.e. endocytosis of particles. Further, we have also identified the cellular uptake of 20-30 nm particles is by enclosing those CNPs in membrane vesicles in caveolae-mediated endocytosis mechanism. In summary, these results indicate that the nanoparticles-cell interaction has pronounced influence on the shape and size of the nanoparticles. These interactions can be further monitored by real time mechanical property measurement of cell membrane.
Title: Real time monitoring of Cell-Nanoparticles interaction and tracking internalization process by mechanical probing using Atomic Force Microscopy.
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Name(s): Ly, Anh, Author
Seal, Sudipta, Committee Chair
Zhai, Lei, Committee Member
Heinrich, Helge, Committee Member
, Committee Member
University of Central Florida, Degree Grantor
Type of Resource: text
Date Issued: 2014
Publisher: University of Central Florida
Language(s): English
Abstract/Description: With extensive development of nanotechnology in last few years, scientists have discovered that nanoparticles (NPs) can be used as an efficient Drug Delivery System (DOS). In order to develop better NPs based drug delivery tool, it is imperative to understand the interaction between the NPs and the cell membrane. In our earlier studies, cerium oxide nanoparticles (CNPs) have been reported to have therapeutic properties, specifically against abnormalities associated with oxidative stress. Therefore, CNPs with different sizes and morphology were selected to understand the interaction with cell. We analyzed the mechanical property of human nasal septum tumor cells membranes using Atomic Force Microscopy (AFM) with and without CNPs. In particular, Force-Distance spectroscopy mode was used to estimate the elasticity of cells membrane. Different concentrations (0, 50, 125 and 250 ?M) of CNPs were added to the cells (squamous cells; CCL30) and incubated for different time periods (0, 15, 30 and 60 minutes). Cell membrane elasticity/Young's modulus was calculated using a modified Hertz model. Changes in the cell elasticity were observed in high concentration of CNPs when treated with one hour. Significant changes in cell elasticity were observed at high concentration of CNPs for one hour of incubation. No significant change in cell elasticity was observed over one hour time period for 50 ?M of CNPs. Moreover, by using selected inhibitors to block different cell mediated internalization pathways, we also investigated the correlation between the cellular uptake and the tracking of NPs with their size. Specifically, similar change in cell elasticity was observed after blocking the cell energy production for CNPs with smaller diameter (3-5 nm). On the other hand, bigger size NPs (20-30 nm) showed no change in cell elasticity after blocking the cell energy production. These results indicate that 3-5 nm particles internalize cell by non-energy dependent pathway i.e. passive diffusion whereas 20-30 nm particles entered in cell by energy dependent pathways i.e. endocytosis of particles. Further, we have also identified the cellular uptake of 20-30 nm particles is by enclosing those CNPs in membrane vesicles in caveolae-mediated endocytosis mechanism. In summary, these results indicate that the nanoparticles-cell interaction has pronounced influence on the shape and size of the nanoparticles. These interactions can be further monitored by real time mechanical property measurement of cell membrane.
Identifier: CFE0005204 (IID), ucf:50637 (fedora)
Note(s): 2014-05-01
M.S.M.S.E.
Engineering and Computer Science, Materials Science Engineering
Masters
This record was generated from author submitted information.
Subject(s): AFM
Persistent Link to This Record: http://purl.flvc.org/ucf/fd/CFE0005204
Restrictions on Access: campus 2015-05-15
Host Institution: UCF

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