ABSTRACT
Over the last few years, considerable efforts are taken, in order to find a molecular fluorescent probe fulfilling their applicability requirements. Due to a good optical properties and affinity to biological structures conjugated oligoelectrolytes (COEs) can be considered as a promising dyes for application in fluorescence-based bioimaging. In this work, we synthetized COEs with phenylenevinylene core (PV-COEs) and applied as fluorescent membranous-specific probes. Cytotoxicity effects of each COE were probed on cancerous and non-cancerous cell types and little to no toxicity effects were observed at the high range of concentrations. The intensity of cell fluorescence following the COE staining was determined by the photoluminescence analysis and fluorescence activated cell sorting method (FACS). Intercalation of tested COEs into mammalian cell membranes was revealed by fluorescent and confocal microscopy colocalization with commercial dyes specific for cellular structures including mitochondria, Golgi apparatus and endoplasmic reticulum. The phenylenevinylene conjugated oligoelectrolytes have been found to be suitable for fluorescent bioimaging of mammalian cells and membrane-rich organelles. Due to their water solubility coupled with spontaneous intercalation into cells, favorable photophysical features, ease of cell staining, low cytotoxicity and selectivity for membranous structures, PV-COEs can be applied as markers for fluorescence imaging of a variety of cell types.
Subject(s)
Electrolytes/chemistry , Fluorescent Dyes/chemistry , Microscopy, Confocal , Animals , BALB 3T3 Cells , Cell Line , Cell Membrane/chemistry , Cell Membrane/metabolism , Cell Survival , Electrolytes/toxicity , Flow Cytometry , Fluorescent Dyes/toxicity , HeLa Cells , Humans , Intercalating Agents/chemistry , K562 Cells , MiceABSTRACT
OBJECTIVES: Metal oxide nanoparticles (ZnO-NPs and Al2O3-NPs) are used in many fields, including consumer products and biomedical applications. As a result, exposure to these NPs is highly frequent, however, no conclusive information on their potential cytotoxicity and genotoxicity mechanisms are available. For this reason, we studied cytotoxic and genotoxic effects of ZnO-NPs and Al2O3-NPs on human peripheral blood lymphocytes. MATERIALS AND METHODS: We obtained our goals by using MTT assay, Annexin V-FITC flow cytometry, and alkaline, neural and pH 12.1 versions of comet assay. RESULTS: Exposure of lymphocytes to both NPs for 24 h slightly decreased viability of lymphocytes at ≥ 0.5 mM. For the first time, we revealed using the comet assays that both ZnO-NPs and Al2O3-NPs caused a concentration-dependent increase of DNA single-strand breaks, but not alkali-labile sites. Treatment with DNA glycosylases showed that the NPs induced oxidative DNA damage. DNA damage caused by both nanoparticles at 0.05 mM was removed within 120 min, however lymphocytes did not repair DNA damage induced by 0.5 mM NPs. Studied nanoparticles did not induce apoptosis in lymphocytes. CONCLUSION: Our results suggest that ZnO-NPs and Al2O3-NPs at concentration up to 0.5 mM did not exhibit cytotoxic effect but may exert genotoxic effect on lymphocytes, at least partially by the generation of oxidative DNA damage and strand breaks.
Subject(s)
Aluminum Oxide/toxicity , Apoptosis/drug effects , DNA Damage , Lymphocytes/drug effects , Metal Nanoparticles/toxicity , Oxidative Stress/drug effects , Zinc Oxide/toxicity , Adult , Cell Survival/drug effects , Cells, Cultured , Comet Assay , Consumer Product Safety , DNA Breaks, Single-Stranded , DNA Repair/drug effects , Drug Delivery Systems , Humans , Kinetics , Oxidation-Reduction , Toxicity Tests , Young AdultABSTRACT
Conjugated phenylenevinylene oligoelectrolytes, which consist of a phenylenevinylene core equipped at each end with hydrophilic pendent groups, are shown to be good candidates for mammalian cell membrane staining. When used in the micromolar concentration range, they express low to moderate cell toxicity for selected regular and cancerous cell lines as tested for adherent and suspension cells.
