Single-walled carbon nanotube-induced mitotic disruption.

Carbon nanotubes were among the earliest products of nanotechnology and have many potential applications in medicine, electronics, and manufacturing. The low density, small size, and biological persistence of carbon nanotubes create challenges for exposure control and monitoring and make respiratory exposures to workers likely. We have previously shown mitotic spindle aberrations in cultured primary and immortalized human airway epithelial cells exposed to 24, 48 and 96 µg/cm(2) single-walled carbon nanotubes (SWCNT). To investigate mitotic spindle aberrations at concentrations anticipated in exposed workers, primary and immortalized human airway epithelial cells were exposed to SWCNT for 24-72 h at doses equivalent to 20 weeks of exposure at the Permissible Exposure Limit for particulates not otherwise regulated. We have now demonstrated fragmented centrosomes, disrupted mitotic spindles and aneuploid chromosome number at those doses. The data further demonstrated multipolar mitotic spindles comprised 95% of the disrupted mitoses. The increased multipolar mitotic spindles were associated with an increased number of cells in the G2 phase of mitosis, indicating a mitotic checkpoint response. Nanotubes were observed in association with mitotic spindle microtubules, the centrosomes and condensed chromatin in cells exposed to 0.024, 0.24, 2.4 and 24 µg/cm(2) SWCNT. Three-dimensional reconstructions showed carbon nanotubes within the centrosome structure. The lower doses did not cause cytotoxicity or reduction in colony formation after 24h; however, after three days, significant cytotoxicity was observed in the SWCNT-exposed cells. Colony formation assays showed an increased proliferation seven days after exposure. Our results show significant disruption of the mitotic spindle by SWCNT at occupationally relevant doses. The increased proliferation that was observed in carbon nanotube-exposed cells indicates a greater potential to pass the genetic damage to daughter cells. Disruption of the centrosome is common in many solid tumors including lung cancer. The resulting aneuploidy is an early event in the progression of many cancers, suggesting that it may play a role in both tumorigenesis and tumor progression. These results suggest caution should be used in the handling and processing of carbon nanotubes.

Morphological changes in the hypothalamic arcuate nucleus and median eminence in the golden hamster during the neonatal period.

The purpose of the present investigation was to study the ultrastructure of the arcuate nucleus (ARC) and median eminence of hamsters on days 1-15 of the neonatal period. From days 1-6, the neurons of the ARC had large nuclei and a small amount of cytoplasm which contained polysomes, mitochondria, RER, lysosomes and Golgi complexes. From days 7-15 there was an increase in the amount of cytoplasm as well as more extensive Golgi complexes and RER. Astrocytes were the predominant glial component in both the ARC and median eminence. Astrocytic processes were in juxtaposition to unmyelinated axons, dendrites, and synapses. Axodendritic and axosomatic synapses containing clear vesicles were observed in the neuropil on day 1. There was an increase in the number of dense-core vesicles in the axonal endings beginning on day 4. Concomitantly, there were increasing numbers of clear and dense-core vesicles (64-70 nm) in terminals of the external layer of the median eminence, whereas larger dense-core vesicles (105-140 nm) were distinguishable by day 10 immediately dorsal to the external layer. The capillaries of the median eminence were composed of nonfenestrated endothelium from days 1-9. Fenestrae began to appear about day 10. Ependymal cells lining the third ventricle had pinocytotic vesicles, microvilli, and bleb-like protrusions on their apical surfaces. Ependymal processes were adjacent to nerve processes in the neuropil of the ARC and in the external layer of the median eminence, where they contacted the perivascular space. Two types of supraependymal cells were seen in animals throughout the neonatal period. One resembled a neuron which sent processes along the ependymal surface and often between cells. The second type was similar to a macrophage. The results of this study demonstrate the maturation of the neural elements in the ARC/median eminence area of the neonatal hamster.