Over the past few years, Quantum Dots have been tested in most biotechnological applications that use fluorescence, including DNA array technology, immunofluorescence assays, cell and animal biology. Quantum Dots tend to be brighter than conventional dyes, because of the compounded effects of extinction coefficients that are an order of magnitude larger than those of most dyes. Their main advantage resides in their resistance to bleaching over long periods of time (minutes to hours), allowing the acquisition of images that are crisp and well contrasted. This increased photostability is especially useful for three-dimensional (3D) optical sectioning, where a major issue is bleaching of fluorophores during acquisition of successive z-sections, which compromises the correct reconstruction of 3D structures. The long-term stability and brightness of Quantum Dots make them ideal candidates also for live animal targeting and imaging. The vast majority of the papers published to date have shown no relevant effects on cells viability at the concentration used for imaging applications; higher concentrations, however, caused some issues on embryonic development. Adverse effects are due to be caused by the release of cadmium, as surface PEGylation of the Quantum Dots reduces these issues. A recently published paper shows evidences of an epigenetic effect of Quantum Dots treatment, with general histones hypoacetylation, and a translocation to the nucleus of p53. In this study, mice treated with Quantum Dots for imaging purposes were analyzed to investigate the impact on protein expression and networking. Differential monoand bidimensional electrophoresis assays were performed, with the individuation of differentially expressed proteins after intravenous injection and imaging analysis; further, as several authors indicate an increase in reactive oxygen species as a possible mean of damage due to the Quantum Dots treatment, we investigated the signalling pathway of APE1/Ref1, a protein involved in the response to oxidative stress. Our results, although preliminary, suggest several interesting point of discussion on Quantum Dots imaging for in vivo diagnostic application, but also for a new therapeutic approach.
Molecular and biochemical methods to assess the biological impact of Quantum Dots
CELLINI, Barbara;CALDERAN, Laura;BOSCHI, Federico;SBARBATI, Andrea;OSCULATI, Francesco
2009-01-01
Abstract
Over the past few years, Quantum Dots have been tested in most biotechnological applications that use fluorescence, including DNA array technology, immunofluorescence assays, cell and animal biology. Quantum Dots tend to be brighter than conventional dyes, because of the compounded effects of extinction coefficients that are an order of magnitude larger than those of most dyes. Their main advantage resides in their resistance to bleaching over long periods of time (minutes to hours), allowing the acquisition of images that are crisp and well contrasted. This increased photostability is especially useful for three-dimensional (3D) optical sectioning, where a major issue is bleaching of fluorophores during acquisition of successive z-sections, which compromises the correct reconstruction of 3D structures. The long-term stability and brightness of Quantum Dots make them ideal candidates also for live animal targeting and imaging. The vast majority of the papers published to date have shown no relevant effects on cells viability at the concentration used for imaging applications; higher concentrations, however, caused some issues on embryonic development. Adverse effects are due to be caused by the release of cadmium, as surface PEGylation of the Quantum Dots reduces these issues. A recently published paper shows evidences of an epigenetic effect of Quantum Dots treatment, with general histones hypoacetylation, and a translocation to the nucleus of p53. In this study, mice treated with Quantum Dots for imaging purposes were analyzed to investigate the impact on protein expression and networking. Differential monoand bidimensional electrophoresis assays were performed, with the individuation of differentially expressed proteins after intravenous injection and imaging analysis; further, as several authors indicate an increase in reactive oxygen species as a possible mean of damage due to the Quantum Dots treatment, we investigated the signalling pathway of APE1/Ref1, a protein involved in the response to oxidative stress. Our results, although preliminary, suggest several interesting point of discussion on Quantum Dots imaging for in vivo diagnostic application, but also for a new therapeutic approach.File | Dimensione | Formato | |
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