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Alexandra AP Mansur,1 Herman S Mansur,1 Sandhra M de Carvalho,1–3 Zélia IP Lobato,2 Maria IMC Guedes,2 Maria F Leite3 1Center of Nanoscience, Nanotechnology, and Innovation-CeNano2I, Department of Metallurgical and Materials Engineering, 2Department of Preventive Veterinary Medicine, Veterinary School, 3Department of Physiology and Biophysics, ICB, Federal University of Minas Gerais, Belo Horizonte, MG, Brazil Abstract: Herein, for the first time, we demonstrated that novel biofunctionalized semiconductor nanomaterials made of Cd-containing fluorescent quantum dot nanoconjugates with the surface capped by an aminopolysaccharide are not biologically safe for clinical applications. Conversely, the ZnS-based nanoconjugates proved to be noncytotoxic, considering all the parameters investigated. The results of in vitro cytotoxicity were remarkably dependent on the chemical composition of quantum dot (CdS or ZnS), the nature of the cell (human cancerous and embryonic types), and the concentration and time period of exposure to these nanomaterials, caused by the effects of Cd2+ on the complex nanotoxicity pathways involved in cellular uptake. Unexpectedly, no decisive evidence of nanotoxicity of CdS and ZnS conjugates was observed in vivo using intravenous injections in BALB/c mice for 30days, with minor localized fluorescence detected in liver tissue specimens. Therefore, these results proved that CdS nanoconjugates could pose an excessive threat for clinical applications due to unpredicted and uncorrelated in vitro and in vivo responses caused by highly toxic cadmium ions at biointerfaces. On the contrary, ZnS nanoconjugates proved that the “safe by design” concept used in this research (ie, biocompatible core–shell nanostructures) could benefit a plethora of applications in nanomedicine and oncology. Keywords: fluorescent nanoparticles, semiconductor quantum dots, nanotoxicity, bionanoconjugates, nanoprobes