Cell cycle-dependent endocytosis of DNA-wrapped single-walled carbon nanotubes by neural progenitor cells.

Abstract

While exposure of C17.2 neural progenitor cells (NPCs) to nanomolar concentrations of carbon nanotubes (NTs) yields evidence of cellular substructure reorganization and alteration of cell division and differentiation, the mechanisms of NT entry are not understood. This study examines the entry modes of (GT)₂₀ DNA-wrapped single-walled carbon nanotubes (SWCNTs) into NPCs. Several endocytic mechanisms were examined for responsibility in nanomaterial uptake and connections to alterations in cell development via cell-cycle regulation. Chemical cell-cycle arrest agents were used to synchronize NPCs in early G₁, late G₁/S, and G₂/M phases at rates (>80%) aligned with previously documented levels of synchrony for stem cells. Synchronization led to the highest reduction in SWCNT internalization during the G₁/S transition of the cell cycle. Concurrently, known inhibitors of endocytosis were used to gain control over established endocytic machineries (receptor-mediated endocytosis (RME), macropinocytosis (MP), and clathrin-independent endocytosis (CIE)), which resulted in a decrease in uptake of SWCNTs across the board in comparison with the control. The outcome implicated RME as the primary mechanism of uptake while suggesting that other endocytic mechanisms, though still fractionally responsible, are not central to SWCNT uptake and can be supplemented by RME when compromised. Thereby, endocytosis of nanomaterials was shown to have a dependency on cell-cycle progression in NPCs.