Single-walled carbon nanotubes (SWCNTs) are increasingly being investigated for biomedical imaging, sensing, and drug delivery. Cell types, cellular entry mechanisms, and SWCNT lengths dictate SWCNT uptake, subsequent intracellular trafficking, and retention. Specialized immune cells known as macrophages are capable of two size-dependent entry mechanisms: endocytosis of small particles (diameter < 200 nm) and phagocytosis of large particles (diameter > 500 nm). In comparison, fibroblasts uptake particles predominantly through endocytosis. We report dependence of cellular processing including uptake, subcellular distribution, and retention on the SWCNT length and immune cell-specific processes. We chose SWCNTs of three different average lengths: 50 nm (ultrashort, US), 150 nm (short) and 500 nm (long) to encompass two different entry mechanisms, and noncovalently dispersed them in water, cell culture media, and phosphate buffer (pH 5) with bovine serum albumin, which maintains the SWCNT optical properties and promotes their cellular uptake. Using confocal Raman imaging and spectroscopy, we quantified cellular uptake, tracked the intracellular dispersion state ( i.e. , individualized versus bundled), and monitored recovery as a function of SWCNT lengths in macrophages. Cellular uptake of SWCNTs increases with decreasing SWCNT length. Interestingly, short-SWCNTs become highly bundled in concentrated phase dense regions of macrophages after uptake and most of these SWCNTs are retained for at least 24 h. On the other hand, both US- and long-SWCNTs remain largely individualized after uptake into macrophages and are lost over a similar elapsed time. After uptake into fibroblasts, however, short-SWCNTs remain individualized and are exocytosed over 24 h. We hypothesize that aggregation of SWCNTs within macrophages but not fibroblasts may facilitate the retention of SWCNTs within the former cell type. Furthermore, the differential length-dependent cellular processing suggests potential applications of macrophages as live cell carriers of SWCNTs into tumors and regions of inflammation for therapy and imaging.
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Photoporation Using Carbon Nanotubes for Intracellular Delivery of Molecules and Its Relationship to Photoacoustic Pressure
Exposure of carbon‐black (CB) nanoparticles to near‐infrared nanosecond‐pulsed laser energy can cause efficient intracellular delivery of molecules by photoporation. Here, cellular bioeffects of multi‐walled carbon nanotubes (MWCNTs) and single‐walled carbon nanotubes (SWCNTs) are compared to those of CB nanoparticles. In DU145 prostate‐cancer cells, photoporation using CB nanoparticles transitions from (i) cells with molecular uptake to (ii) nonviable cells to (iii) fragmented cells with increasing laser fluence, as seen previously. In contrast, photoporation with MWCNTs causes uptake and, at higher fluence, fragmentation, but does not generate nonviable cells, and SWCNTs show little evidence of bioeffects, except at extreme laser conditions, which generate nonviable cells and fragmentation, but no significant uptake. These different behaviors cannot be explained by photoacoustic pressure output from the particles. All particle types emit a single, ≈100 ns, mostly positive‐pressure pulse that increases in amplitude with laser fluence. Different particle types emit different peak pressures, which are highest for SWCNTs, followed by CB nanoparticles and then MWCNTs, which does not correlate with cellular bioeffects between different particle types. This study concludes that cellular bioeffects depend strongly on the type of carbon nanoparticle used during photoporation and that photoacoustic pressure is unlikely to play a direct mechanistic role in the observed bioeffects.
more » « less- NSF-PAR ID:
- 10047637
- Publisher / Repository:
- Wiley Blackwell (John Wiley & Sons)
- Date Published:
- Journal Name:
- Advanced Healthcare Materials
- Volume:
- 7
- Issue:
- 5
- ISSN:
- 2192-2640
- Format(s):
- Medium: X
- Sponsoring Org:
- National Science Foundation
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