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||Assessment of nanoparticles toxicity by flow cytometry using human erythrocytes
||Tuşa I., Rugina A., Dobre A.-M., Ciotec A., Calu L., Stan L., Dumitrache F., Fleaca C.,Morjan I, Cotoraci C., Bratosin D.
||1National Institute for Biological Science Research and Development, Biotechnological center for cell therapy and regenerative medicine based on stem cell and apoptosis modulators” (BIOREGMED) Bucharest, Romania
2 The National Institute for Laser, Plasma and Radiation Physics, Bucharest, Romania
3”Vasile Goldis” Western University of Arad, Faculty of Medicine, Pharmacy and Dental Medicine, Arad,Romania
4”Vasile Goldis” Western University of Arad, Faculty of Natural Sciences, Arad, Romania
||The use of nanoparticles in biological and medical applications has increased in the last years. Understanding their potential toxicity and effect on the body is crucial before clinical use. The most used cell culture systems to test their cytotoxicity are of nonhuman origin. For assessing cytotoxicity of nanoparticles we developed a new experimental cell system, the RBCs, and we have evaluated the toxic effects by flow cytometric analysis. Recently, we and other researchers have shown that PCD of human erythrocyte is related to an apoptosis mechanism operating in the absence of mitochondria and lacking the terminal effectors of nuclear apoptosis (erythroptosis also called eryptosis) and to determine human RBC viability, we described a new rapid and sensitive calcein-AM assay by flow cytometric determination. Consequently, in the present study, to evaluate cell-nanoparticles interactions, human RBCs were exposed to different concentrations of nanoparticles and analyzed by flow cytometry, after 3h and 24h incubation endpoints for morphological changes (FSC/SSC) and viability (calcein-AM method). Our results indicate that flow cytometric analysis of RBCs viability using calcein-AM discrimination could provide a rapid and accurate experimental cellular model for effectively screening and evaluating biological responses for in vitro nanotoxicology.
||human erythrocyte, cell viability, flow cytometric analysis, nanotoxicity, nanoparticles.
||Daniela Bratosin, National Institute for Biological Science Research and Development, Biotechnological center for cell therapy and regenerative medicine based on stem cell and apoptosis modulators” (BIOREGMED) Bucharest, Romania Tel/Fax +40-(021)-2200881, email: firstname.lastname@example.org.