Publication details

Monitoring of cell adhesion and cytotoxicity effect of derivative vitamine E using a piezoelectric biosensor



Year of publication 2006
Type Article in Proceedings
Conference The 9th World Congress on Biosensors - Delegate Manual
MU Faculty or unit

Faculty of Science

Field Biochemistry
Keywords Piezoelectric biosensor; Resistance; Cytotoxicity assay
Description The quartz crystal microbalance (QCM) was used to construct a piezoelectric biosensor utilizing living rat epithelial cells WB F344 and lung melanoma cells B16F10 as biorecognition elements. The resonant frequency delta-f and resistance delta-R changes were measured during adhesion of the cells on the gold surfaces modified by the layers of either hydrophobic polystyrene or extracellular matrix proteins as fibronectin, laminin and vitronectin. The piezoelectric biosensor was furthermore utlized for monitoring of apoptosis initiated by the addition of vitamin E derivatives as a-tocopheryl amidomalate (a-TAM). Next, the impedance scans were used to followed the conductivity of gap junctions of the WB F344 cell line before and after the cytotoxic effects of several anticancer drugs. The 10 MHz quartz crystal with the gold electrodes on the both sides was placed at the bottom of the thermostated measurement chamber. The cells adhered on the gold surface were identified by staining with MitoTracker Red fluorescent dye. The concentration of a-TAM added to the cell medium was 300 uM. The process on the sensor was simultaneously observed and compared on microplates. The initial cell attachment and spreading induced a decrease of frequency and increase of resistance. The steady-state of both shifts was achieved after a few hours. The presence of cells on the surface was confirmed by fluorescent microscopy. The apoptosis was morphologically characterized by severe alterations in the cell shape like shrinkage and disintegration of cell-cell contacts and therefore it was sensitively measured by QCM. Consequently, the decreased frequency and increased resistance values were observed after the addition of a-TAM. The information from the simultaneous on-line monitoring of both f and R changes revealed viscoelastic properties of the cell monolayer on the sensing surface. As the attached cells formed an insulating layer, the observation of the conductivity of the gap junction after the drug treatment was shown as the increase of the conductivity. The results indicate that the piezoelectric biosensor is suitable for real-time studies of cell adhesion and for identification and screening of biologically active drugs and other biomolecules affecting cellular shape and attachment.
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