Sviluppo di un nuovo sistema per la discriminazione tra il consumo di ossigeno e la variazione del flusso ematico regionale cerebrale

Aim:The possibility to discern between the oxygen consumption and regional blood flow, on specific cerebral district, would offer considerable advantages from the point of view of the in-vivo study of the energetic metabolism of the CNS, both in physiological conditions and in animal models of neurodegenerative diseases such as Parkinson's disease.Methods:the process of research and development, which involved different contexts and a multidisciplinary approach, led to the creation of a miniaturized prototype, fully portable, that combines the production of hydrogen and oxygen in physiological environment to the their concomitant reading. Oxygen microsensor has been attained modifying a previously developed O2sensor, a new microsensor for hydrogen detection was obtained by means of platinum electrodeposition on graphite and nano carbon particles. Hydrogen and oxygen used for sensors characterization and for in-vitro studies on cell cultures (PC12), have been produced in situ by electrolysis of a buffered solution at physiological pH using platinum microelectrodes. In-vitro experiments were done by coupling the previously described sensors with a microdialysis device used for cellular cultures (to simulate blood flow) and measuring gases levels through the technique of clearance of hydrogen and oxygen.Results:A decay of both signals was been observed. The oxygen decay has been related with both cellular metabolism and microdialysis flow, while the hydrogen signal was only dependent on the rate of perfusion. The enrichment with CNT resulted in a higher sensitivity of the oxygen sensor and a good sensitivity and stability for hydrogen sensor. PC12 oxygen consumption and its real-time fluctuations were measured, even after a pharmacological treatment targeting mitochondrial functions. With a constant perfusion flow, hydrogen clearance didn’t show statistically significant changes.Conclusion:The developed device has allowed the discrimination between oxygenin-vitroutilization and perfusion flow.