Multi-walled carbon nanotubes (MWNTs) pose a respiratory hazard because they can cause pulmonary fibrosis, which may lead to mesothelioma. The Bionanosciences Group at UT Dallas is interested in understanding the interactions of triblock polymer Pluronic® F108-coated MWNTs with macrophages that are the first responders to nanoparticles encountered in the body. This information is relevant for improving the biomedical efficacy of MWNT therapeutics, for understanding mechanisms of MWNT biocompatibility, and for developing methods to ameliorate MWNT toxicity. To better understand mechanisms of MWNT toxicity, it is important to know whether MWNTs physically enter cells and where they locate inside cells. We have developed procedures to detect internalized MWNTs and reconstruct 3D images of RAW 264.7 mouse macrophages with cell-associated MWNTs by laser scanning confocal Raman microscopy. 3D images of cells are reconstructed with stacks of optical sections from confocal planes to place the subcellular MWNT locations in the context of the intact cell. The 3D MWNT Raman images demonstrated that Pluronic® F108-coated pristine MWNTs (P-MWNTs) and carboxylated MWNTs (C-MWNTs) exposed to RAW 264.7 cells for 24 h at 37 °C accumulated inside the cells within punctate vesicles, most likely in the endosome/lysosome system, but not in the cytoplasm. These results are consistent with our previous observations that RAW 264.7 cells accumulate ~100 times more C-MWNTs suspended in the surfactant Pluronic® F-108 than corresponding P-MWNTs during a 24 h incubation at 37 °C. Future work will involve assessing the intracellular distributions of various functionalized MWNTs and evaluating their fate after internalization, and whether they are degraded, secreted, or if they persist within cells. Also, Raman imaging with live cells will be used to access whether accumulation of MWNTs induce the release of cytokines that may cause lysosomal membrane damage and result in the redistribution of MWNTs to the cytoplasm.

Label-free three-dimensional Raman imaging of carbon nanotubes in mammalian cells

Macromolecular Chemistry: The Second Century

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ACS Spring 2020 National Meeting & Expo

The official archive and repository for all ACS Meetings & Expositions abstracts and select posters or presentations.