Development of nano-scale observation system for adhesion and uptake of potholes on/into living cells
| Project/Area Number |
18360394
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Biofunction/Bioprocess
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| Research Institution | Kyoto University |
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Principal Investigator |
HIGAHITANI Ko Kyoto University, Dpt. of Chem. Eng, Professor (10039133)
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| Co-Investigator(Kenkyū-buntansha) |
SHINTO Hiroyuki Kyoto Univ, Dpt. of Chem. Eng, Assistant Prof (80324656)
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| Project Period (FY) |
2006 – 2007
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| Project Status |
Completed (Fiscal Year 2007)
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| Budget Amount *help |
¥16,800,000 (Direct Cost: ¥15,900,000、Indirect Cost: ¥900,000)
Fiscal Year 2007: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000)
Fiscal Year 2006: ¥12,900,000 (Direct Cost: ¥12,900,000)
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| Keywords | Nan biology / Scanning probe microscopy / Confocal laser scanning microscopy / Surface / interface property / Cells and tissues / Nanoparticles / Adhesion / Uptake |
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| Research Abstract |
We developed an integrated nano-scale observation system of an Atomic Force Microscope and a Confocal Laser Scanning Microscope (CLSM) with the functionalized colloid AFM probes. This system has been used to obtain the following information: (1) adhesion forces between engineered materials and living cells; (2) optimal size and surface property of particles for cellular uptake; (3) processes of cellular uptake and discharge; (4) movement of the target cells and their neighboring cells. Our purpose is to obtain the quantitative fundamental data for efficient design for DOS particles using our developed system and to understand the mechanisms of adhesion, uptake, and cytotoxicity of particles for living cells.
First, we carried out AFM measurement of the adhesion forces between mouse skin cancer cells (B16F10) and particles with different surface properties of charge density, hydorophilicity/hydrophobicity, and functional group. Second, we performed CLSM observation of the living cells after exposure to culture media including fluorescent nanoparticles with various surface functionalities and sizes. Also, we assessed the cytotoxicity of the fluorescent nanoparticles using the trypan blue dye method.
It was found that the surface functionality of nanoparticles affects the uptake region of cells (the outer surface of cell membrane or the cytoplasm) to determine the cytotoxicity. These uptake region and cytotoxicity were affected by the culture conditions such as temperature and serum addition. There were no clear relationship between the uptake amount and the adhesion forces, indicating that the cellular uptake of particles is not satisfactorily explained by the adhesion forces and additional factor(s) should be therefore took into account.
In the future study, we should carry out the experiments focusing on the receptors residing in/on cell membrane (i.e., the membrane proteins)▲to understand the detailed mechanisms of cellular uptake of particles.
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Report
(3 results)
Research Products
(58 results)
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[Journal Article] Chemical groups that adhere to the surfaces of living malignant cells2007
Author(s)
C.E. McNamee, Y., Aso, S., Yamamoto, Y., Fukumori, H., Ichikawa, K., Higashitani
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Journal Title
Pharmaceutical Research 24
Pages: 2370-2380
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Effect of the cell type and cell density on the binding of living cells to a silica particle : An atomic force microscope study2006
Author(s)
N., Pyo, S., Tanaka, C.E., McNamee, Y., Kanda, Y., Fukumori, H., Ichikawa, K., Higashitani
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Journal Title
Colloid Surfaces 53
Pages: 278-287
Description
「研究成果報告書概要(欧文)」より
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[Presentation] AFM/CLSM Study of Adhesion and Uptake of Particles on/by Melanoma Cells2007
Author(s)
H., Shinto, Y., Aso, J., Tsujimura, Y., Ohta, K., Higashitani
Organizer
Nano-interface workshop "Recent progress in Japan"
Place of Presentation
University of South Australia, Adelaide
Year and Date
2007-12-07
Description
「研究成果報告書概要(欧文)」より
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[Presentation] Adhesion of particles on cancer cells and the size effect (in Japanese)2007
Author(s)
H., Shinto, Y., Aso, J., Tsujimura, C.E., McNamee, K., Higashitani
Organizer
39th autumn meeting of the Society of Chemical Engineers, (Japan)
Place of Presentation
Hokkaido University
Year and Date
2007-09-15
Description
「研究成果報告書概要(欧文)」より
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[Presentation] AFM/LSM study of adhesion and uptake of particles on/into living cells2007
Author(s)
H., Shinto, Y., Aso, J., Tsujimura, C.E., McNamee, K., Higashitani
Organizer
Summer school of "Colloid interface phenomena in soil, water, and life environment"
Place of Presentation
Tsukuba University
Year and Date
2007-08-25
Description
「研究成果報告書概要(欧文)」より
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[Presentation] Adhesion and Uptake of Particles on/by Living Cells and the Cytotoxicity2007
Author(s)
H., Shinto, Y., Aso, J., Tsujimura, Y., Ohta, K., Higashitani
Organizer
Core-to-Core : Young Researchers Symposium
Place of Presentation
The University of Melbourne, Australia
Description
「研究成果報告書概要(欧文)」より
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