title: Cytotoxicity of water-soluble fullerene in vascular endothelial cells.
abstract: Nanoscale materials are presently under development for diagnostic (nanomedicine) and electronic purposes. In contrast to the potential benefits of nanotechnology, the effects of nanomaterials on human health are poorly understood. Nanomaterials are known to translocate into the circulation and could thus directly affect vascular endothelial cells (ECs), causing vascular injury that might be responsible for the development of atherosclerosis. To explore the direct effects of nanomaterials on endothelial toxicity, human umbilical vein ECs were treated with 1-100 microg/ml hydroxyl fullerene [C60(OH)24; mean diameter, 7.1 +/- 2.4 nm] for 24 h. C60(OH)24 induced cytotoxic morphological changes such as cytosolic vacuole formation and decreased cell density in a dose-dependent manner. Lactate dehydrogenase assay revealed that a maximal dose of C60(OH)24 (100 microg/ml) induced cytotoxic injury. Proliferation assay also showed that a maximal dose of C60(OH)24 inhibited EC growth. C60(OH)24 did not seem to induce apoptosis but caused the accumulation of polyubiquitinated proteins and facilitated autophagic cell death. Formation of autophagosomes was confirmed on the basis of Western blot analysis using a specific marker, light chain 3 antibody, and electron microscopy. Chronic treatment with low-dose C60(OH)24 (10 microg/ml for 8 days) inhibited cell attachment and delayed EC growth. In the present study, we have examined, for the first time, the toxicity of water-soluble fullerenes to ECs. Although fullerenes changed morphology in a dose-dependent manner, only maximal doses of fullerenes caused cytotoxic injury and/or death and inhibited cell growth. EC death seemed to be caused by activation of ubiquitin-autophagy cell death pathways. Although exposure to nanomaterials appears to represent a risk for cardiovascular disorders, further in vivo validations are necessary.
authors.affiliation: Dept. of Epidemiology, Research Institute, National Cardiovascular Center, Suita, Osaka 565-8565, Japan.
authors.forename: Hideyuki
authors.initials: H
authors.lastname: Yamawaki
authors.forename: Naoharu
authors.initials: N
authors.lastname: Iwai
mesh_terms: D001343:Autophagy / Q000187:drug effects; D015153:Blotting, Western; D049109:Cell Proliferation / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D042783:Endothelial Cells / Q000187:drug effects* / Q000648:ultrastructure*; D037741:Fullerenes / Q000494:pharmacology*; D006801:Humans; D046529:Microscopy, Electron, Transmission; D049329:Nanostructures / Q000009:adverse effects*; D014471:Umbilical Veins / Q000166:cytology
chemical_list: D037741:Fullerenes