Assessment of oxidative DNA damage and repair at single cellular level via real-time monitoring of 8-OHdG biomarker.
8-Hydroxydeoxyguanosine (8-OHdG) is the most important and best-documented biomarker of oxidative stress, which is involved in the instigation of various diseases. 8-OHdG levels correlate to oxidative DNA damage which is known to be the root cause of a variety of age-related chronic diseases. The purpose of our research was to develop a detection strategy capable of measuring 8-OHdG in real-time at the surface of a single cell. Activated carbon fiber microelectrodes were used as the sensing platform. The microelectrodes were used to measure 8-OHdG release from single lung epithelial cells under the influence of nicotine. In order to evaluate the direct role of nicotine in tobacco induced genotoxicity, we studied the influence of parameters such as nicotine concentration and exposure times on 8-OHdG secretion. 2-8 mM nicotine solutions induced dose-dependent DNA damage in single cells, which was observed via amperometric measurements of secreted 8-OHdG biomarker. Real-time 8-OHdG measurements from single cells exposed to 4 mM nicotine solution revealed cessation of 8-OHdG secretion after 110 min. We have successfully outlined a methodology to detect 8-OHdG at the surface of single cells. A similar protocol can be used to evaluate oxidative DNA damage and repair mechanisms in other disease models.
Prabhulkar
Shradha
S
Nanobioengineering/Bioelectronics Laboratory, Department of Biomedical Engineering, Florida International University, 10555 West Flagler Street, Miami, FL 33174, United States.
Li
Chen-Zhong
CZ
D000080242:8-Hydroxy-2'-Deoxyguanosine; D015415:Biomarkers / Q000032:analysis; D015374:Biosensing Techniques / Q000379:methods*; D002244:Carbon; D000077482:Carbon Fiber; D002478:Cells, Cultured; D003199:Computer Systems; D004249:DNA Damage; D004260:DNA Repair; D003849:Deoxyguanosine / Q000031:analogs & derivatives* / Q000032:analysis; D004847:Epithelial Cells / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D008839:Microelectrodes; D009538:Nicotine / Q000633:toxicity; D010084:Oxidation-Reduction; D011650:Pulmonary Alveoli / Q000166:cytology / Q000187:drug effects / Q000378:metabolism
D015415:Biomarkers; D000077482:Carbon Fiber; D009538:Nicotine; D002244:Carbon; D000080242:8-Hydroxy-2'-Deoxyguanosine; D003849:Deoxyguanosine