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Characterization by Electron Spin Resonance Spectroscopy of Reactive Oxygen Species Generated by Titanium Dioxide and Hydrogen Peroxide

M.-C. Lee^1,*,, F. Yoshino^1,, H. Shoji¹, S. Takahashi¹, K. Todoki¹, S. Shimada¹, and K. Kuse-Barouch^1,2

¹ Department of Clinical Care Medicine, Division of Pharmacology and ESR Laboratories, Kanagawa Dental College, 82 Inaoka-cho, Yokosuka, Kanagawa, 238-8580, Japan; and
² Center for Implantology, Boston University Goldman School of Dental Medicine, 100 East Newton Street, Boston, MA 02118-2392, USA;

View larger version (18K): [in a new window]   Figure 1. Schematic diagram of the experimental protocols used for non-irradiated and irradiated TiO2. Experimental protocol for non-irradiated TiO2 (A) or for irradiated (365 nm) TiO2 (B) in the presence of H2O2 for ESR measurement.

View larger version (25K): [in a new window]   Figure 2. Hydroxyl radical (HO•) generation from TiO2 in the presence of H2O2. (A) The time course of ESR spectrum from TiO2 measured by EPR spin-trapping following the addition of H2O2 at time 0 and with DMPO as a spin trap. (B) Monitoring of the spin concentration of the HO• generated from TiO2 in the presence of H2O2. All data are presented as the mean (n = 3–5). Experimental conditions are described in "MATERIALS & METHODS" and illustrated in Fig. 1.

View larger version (27K): [in a new window]   Figure 3. Superoxide (O2•-) generation from irradiated TiO2 in the presence of H2O2. (A) The time course of ESR spectrum during and after irradiation (365 nm) of TiO2 measured by EPR spin-trapping following the addition of H2O2 at time 0 and with DMPO as a spin trap. (B) Monitoring of the spin concentration of the O2•- generated from irradiated TiO2 (5 min) in the presence of H2O2. Each column represents the mean (n = 5). Experimental conditions are described in "MATERIALS & METHODS" and illustrated in Fig. 1.

View larger version (20K): [in a new window]   Figure 4. Dose-dependent effects of H2O2 on reactive oxygen species (ROS) generation from irradiated TiO2. The ESR spectrum of irradiated (365 nm) TiO2 (0.15%, wt/vol) measured by EPR spin-trapping following the addition of different concentrations of H2O2 (0.0003%–30%) at 1 min and with DMPO (880 mM) as the spin trap.