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ESR technique for noninvasive way to quantify cyclodextrins effect on cell membranes
Authors:A. Grammenos  A. Mouithys-Mickalad  M. Lismont  M. Hoebeke
Affiliation:a Laboratory of Biomedical Spectroscopy, Department of Physics, B5, University of Liège, Sart-Tilman, Belgium
b Center of Oxygen, Research and Development (CORD), Department of Chemistry, B6a, University of Liège, Sart-Tilman, Belgium
c Laboratory of Pharmaceutical Technology, Department of Pharmacy, CHU, B36, University of Liège, 1 Av. de l’Hopital, Belgium
Abstract:A new way to study the action of cyclodextrin was developed to quantify the damage caused on cell membrane and lipid bilayer. The Electron Spin Resonance (ESR) spectroscopy was used to study the action of Randomly methylated-beta-cyclodextrin (Rameb) on living cells (HCT-116). The relative anisotropy observed in ESR spectrum of nitroxide spin probe (5-DSA and cholestane) is directly related to the rotational mobility of the probe, which can be further correlated with the microviscosity. The use of ESR probes clearly shows a close correlation between cholesterol contained in cells and cellular membrane microviscosity. This study also demonstrates the Rameb ability to extract cholesterol and phospholipids in time- and dose-dependent ways. In addition, ESR spectra enabled to establish that cholesterol is extracted from lipid rafts to form stable aggregates. The present work supports that ESR is an easy, reproducible and noninvasive technique to study the effect of cyclodextrins on cell membranes.
Keywords:Cholesterol   Cyclodextrin   ESR   Lipid rafts   Membrane viscosity   Spin label
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