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The influence of reactive oxygen species (ROS) on the surface modification of titanium implants and osseointegration is unclear. The aim of this study was to evaluate the ability of titanium dioxide (TiO2) to generate ROS in the presence of H2O2 and to determine whether any ROS thus generated play a role in osseointegration, as measured by electron spin resonance (ESR) spin-trapping with 5,5-dimethyl-1-pyrolline-N-oxide (DMPO). We demonstrate that TiO2 together with H2O2 generated hydroxyl radicals (HO*), as shown by a time-dependent increase in the spin concentration of the ESR signal for the DMPO-OH spin adduct, indicating HO* generation. Interestingly, irradiated TiO2 with H2O2 generated the superoxide (O2*-), as shown by an increase in the spin concentration of the signal for the DMPO-OOH spin adduct, indicating O2*- generation during the period of irradiation (0-5 min). These results suggest that ROS generated from the TiO2 layer may be involved in creating appropriate conditions for the osseointegration of dental implants into alveolar bone tissues.

A number of researchers have reported that free radicals generated in the brain are involved in various brain dysfunctions, including ischemia-reperfusion injury, brain tumors, and neurodegenerative diseases. It has been reported that the spin probe MC-PROXYL can penetrate the blood-brain barrier and can be useful for evaluating oxidative stress in the brain. Preliminary comparisons were made by ESR imaging of the heads of live mice and isolated rat brains using the spin probe MC-PROXYL and the blood-brain-barrier impermeable probe carbamoyl-PROXYL. The results showed that MC-PROXYL, but not carbamoyl-PROXYL, was widely distributed in the brain. These methods were also applied for the imaging of brains from stroke-prone spontaneously hypertensive rats (SHRSPs). The rapid decay of 2D ESR images of MC-PROXYL in isolated SHRSP-brain was observed, compared to Wistar-Kyoto rats (WKYs), using the ESR imaging system. Furthermore, we provide evidence, by using L-band ESR non-invasively, that the decay rate of MC-PROXYL in the head region is faster in live SHRSPs than in live WKYs. Taken together, the high oxidative stress sustained by oxygen radical generation in SHRSPs may cause the alteration of MC-PROXYL metabolism in the brain. Our results suggest that in vivo ESR could be applied to the assessment of antioxidant effects on oxidative stress in the brain in animal disease models, such as the SHRSP.

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