Study of the effect of ascorbic acid on protection against UV radiation in cells through the development of electrochemical nanosensors for localized and real-time detection

• Grant number: 22/03643-9
• Support Opportunities: Scholarships in Brazil - Doctorate (Direct)
• Start date: December 01, 2022
• Status: Discontinued
• Field of knowledge: Physical Sciences and Mathematics - Chemistry - Analytical Chemistry
• Principal Investigator: Mauro Bertotti
• Grantee: Pedro Henrique Alves Damasceno
• Host Institution: Instituto de Química (IQ). Universidade de São Paulo (USP). São Paulo , SP, Brazil
• Associated research grant: 18/08782-1 - Miniaturized and integrated chemical sensors: new fabrication platforms for biological, clinical and environmental applications, AP.TEM
• Associated scholarship(s): 25/00212-5 - Scanning Electrochemical Microscopy (SECM) and Atomic Force Microscopy (AFM) to study electrochemical and morphological changes related to ascorbate uptake in biological cells., BE.EP.DD

Abstract

UV radiation is one of the leading environmental agents that causes cancer and photo-aging, the process responsible for the formation of wrinkles and blemishes. Several cellular antioxidants, enzymatic and non-enzymatic, are involved in protecting against these processes. Studies point to the important role of ascorbate in protecting against the formation of damage in biomolecules after UV exposure. Knowing the variations in ascorbate concentration before and after exposure to UV radiation, as well as the relationship between these concentrations and lesions in biomolecules, can provide important information about the role of this species in cellular protection. For studies in cellular environments, electrochemical microscopy Scanning (SECM) is a versatile technique that allows obtaining fast information with high spatial resolution. One of the most significant benefits of SECM is the exploration of microscopic domains with reduced sample volume. In this context, the project´s primary goal is to monitor the ascorbate concentration in HaCaT cells in regions close to their surface and interior, before and after being stimulated by UV light irradiation, using the SECM technique and nanosensors with high selectivity. and sensitivity. The levels of 8-oxo-2'-deoxyguanosine and adducts formed by the reaction of DNA with lipid peroxidation products will be measured to evaluate the redox stress in cell. For comparison, oxidized and reduced glutathione levels will also be assessed by LC/MS/MS. Such studies will contribute to understand the profile of vitamin C levels in HaCaT cells (intra and/or extracellular) throughout the oxidative stress induced by UV irradiation. The project is interdisciplinary, involving a group of biochemistry and analytical chemistry aiming to establish a cause and consequence relationship for the cellular disorders and the levels of vitamin C observed, as well as to define a recommended level of safety of this substance concerning prevention or remediation to cell damage. (AU)