Detection of cellular damage by hydrogen peroxide using SV40-T2 cells on shear horizontal surface acoustic wave (SH-SAW) sensor.
The rat lung epithelial cell line SV40-T2 was used to develop a cellular biosensing system to assay for environmental toxicants. The novel approach on which this system is based involves direct attachment of cultured rat or human cells onto a cell-adhesive matrix on the device through which shear horizontal surface acoustic waves (SH-SAW) are transmitted using 50 MHz SAW resonator. This novel design enables sensitive monitoring of changes of the electrophysical characteristics of cells, such as their conductivity and relative permittivity. A time-dependent change of phase of SAW and change of insertion loss (change of amplitude) were observed when the cells were treated with 0.5 or 1.0 mM H2O2. The change of insertion loss was biphasic, with an early phase (1-3 h) and a late phase (3-6 h). The late phase coincided with the destruction of cell-cell tight junctions detected by measurement of the transepithelial electrical resistance and paracellular permeability; in contrast, the early phase coincided with the destruction of intracellular actin filaments by H2O2. The early-phase effect of H2O2 on phase shift may be attributable to the change of intracellular permittivity by a change of cellular polarity. Immunofluorescence microscopy showed the disappearance of zonula occludens protein 1 from the region of cell-cell contact. These results suggest the correlation between the change of insertion loss as an SAW parameter and the destruction of tight junctions of the cells on the SH-SAW device in the late phase.
Faculty of Science and Technology, Department of Intelligent Machines and System Engineering, Hirosaki University, Japan.
Takumi
T
Higashiyama
Faculty of Agriculture and Life Science, Department of Biochemistry and Molecular Biology, Hirosaki University, 3 Bunkyo-cho, Hirosaki 036-8561, Japan.
Akihiro
A
Katsuyama
Faculty of Science and Technology, Department of Intelligent Machines and System Engineering, Hirosaki University, Japan.
Hideki
H
Otori
Faculty of Agriculture and Life Science, Department of Biochemistry and Molecular Biology, Hirosaki University, 3 Bunkyo-cho, Hirosaki 036-8561, Japan.
Toru
T
Kamimura
Faculty of Science and Technology, Department of Intelligent Machines and System Engineering, Hirosaki University, Japan.
Atsushi
A
Uehara
Faculty of Science and Technology, Department of Intelligent Machines and System Engineering, Hirosaki University, Japan.
Miho
M
Kainuma
Faculty of Agriculture and Life Science, Department of Biochemistry and Molecular Biology, Hirosaki University, 3 Bunkyo-cho, Hirosaki 036-8561, Japan.
Ryo
R
Takumi
Faculty of Agriculture and Life Science, Department of Biochemistry and Molecular Biology, Hirosaki University, 3 Bunkyo-cho, Hirosaki 036-8561, Japan.
Yukako
Y
Kudo
Faculty of Agriculture and Life Science, Department of Biochemistry and Molecular Biology, Hirosaki University, 3 Bunkyo-cho, Hirosaki 036-8561, Japan.
Masayuki
M
Ebina
National Institute for Environmental Studies, 16-2, Onogawa, Tsukuba-shi, Ibaraki 305-8506, Japan.
Katsumi
K
Mochitate
RIVER ELETEC Corporation, 2-1-11 Fujimigaoka, Nirasaki, Yamanashi 407-8502, Japan.
Tasuku
T
Kon
Faculty of Science and Technology, Department of Intelligent Machines and System Engineering, Hirosaki University, Japan.
Yasubumi
Y
Furuya
Faculty of Agriculture and Life Science, Department of Biochemistry and Molecular Biology, Hirosaki University, 3 Bunkyo-cho, Hirosaki 036-8561, Japan. Electronic address: hkikuchi@cc.hirosaki-u.ac.jp.
Hideaki
H
Kikuchi
D000162:Acoustics / Q000295:instrumentation*; D000199:Actins / Q000187:drug effects; D000818:Animals; D015374:Biosensing Techniques / Q000295:instrumentation*; D002460:Cell Line; D002478:Cells, Cultured; D017097:Electric Impedance; D004847:Epithelial Cells / Q000000981:diagnostic imaging* / Q000187:drug effects*; D006861:Hydrogen Peroxide / Q000633:toxicity*; D008856:Microscopy, Fluorescence; D051381:Rats; D033081:Shear Strength; D019108:Tight Junctions / Q000000981:diagnostic imaging / Q000187:drug effects; D014463:Ultrasonography
D000199:Actins; D006861:Hydrogen Peroxide
Epithelial cell; Hydrogen peroxide; Shear horizontal surface acoustic wave; Tight junction; Transepithelial electrical resistance