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. 2007:19:3136-3141.
doi: 10.1002/adma.200701974.

Gold Nanorods Mediate Tumor Cell Death by Compromising Membrane Integrity

Ling Tong, Yan Zhao, Terry B Huff, Matthew N Hansen, Alexander Wei, Ji-Xin Cheng

Gold Nanorods Mediate Tumor Cell Death by Compromising Membrane Integrity

Ling Tong et al. Adv Mater. 2007.
No abstract available

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Figures

Figure 1
Figure 1
Left, Transmission electron micrograph (JEOL 2000FX, 200 kV) of gold nanorods (λmax = 765 nm), prepared by seeded growth method. Right, folate-oligoethyleneglycol ligands, conjugated onto gold nanorod surfaces by in situ dithiocarbamate formation.
Figure 2
Figure 2
Targeted adsorption and uptake of folate-conjugated GNRs (F-NRs, red) by KB cells overexpressing folate receptors (imaged in transmission mode, grey). (A) A high density of F-NRs was observed on the surface of KB cells after 6 h incubation at 37 °C. (B) F-NRs were internalized into KB cells and delivered to the perinuclear region after 17 h incubation. (C) No binding was observed of F-NRs to NIH-3T3 cells, which express folate receptors at a low level. Bar = 10 μm.
Figure 3
Figure 3
Site-dependent photothermolysis mediated by F-NRs (red). (A,B) Cells with membrane-bound F-NRs exposed to cw NIR laser irradiation experienced membrane perforation and blebbing at 6 mW power. The loss of membrane integrity was indicated by EB staining (yellow). (C,D) Cells with internalized F-NRs required 60 mW to produce a similar level of response. (E,F) F-NRs internalized in KB cells labeled by folate-Bodipy (green) were exposed to laser irradiation at 60 mW, resulting in both membrane blebbing and disappearance (melting) of the F-NRs. (G,H) NIH-3T3 cells were unresponsive to F-NRs, and did not suffer photoinduced damage upon 60 mW laser irradiation. (I,J) Cells with membrane-bound F-NRs exposed to fs-pulsed laser irradiation produced membrane blebbing at 0.75 mW. (K,L) Cells with internalized F-NRs remained viable after fs-pulsed irradiation at 4.50 mW, as indicated by a strong calcein signal (green).
Figure 4
Figure 4
F-NR-mediated disruption of actin filaments in actin-GFP (green) transfected KB cells. (A) KB cell with internalized F-NRs (red) before cw laser irradiation. (B) Membrane blebbing accompanied by redistribution of actin-GFP and loss of fluorescence, after an 81.4-s exposure to cw irradiation at 90 mW. (C, D) KB cells without F-NRs, which did not experience membrane blebbing after exposure to cw irradiation at 90 mW. (E) Histogram showing the decrease in actin-GFP fluorescence intensity in cells with and without F-NR labeling (N=5) after cw irradiation. The minor reduction of fluorescence in cells without F-NR labeling is attributed to photobleaching. (F) Blebbing, redistribution of actin-GFP, and loss of fluorescence in KB cells after 2-h treatment with cytochalasin D (5 μg/mL). Bar = 10 μm.
Figure 5
Figure 5
Membrane blebbing is induced by Ca2+ influx during F-NR-mediated photothermolysis. (A,B) Cells with membrane-bound F-NRs (red) in PBS containing 0.9 mM Ca2+ (100 mg/L CaCl2) exhibited blebbing after exposure to fs-pulsed laser irradiation at 3 mW for 61.5 s. (C) Incubation with 2.5 μM EB (red) and 2 μM Oregon Green 488 for 20 min indicated a compromise in membrane integrity and an elevation in intracellular Ca2+, respectively. (D, E) Cells with membrane-bound F-NRs in Ca2+-free PBS were visibly unchanged by fs-pulsed laser irradiation at 3 mW. (F) Incubation with 2.5 μM EB for 15 min revealed perforation of the cell membrane. (G-I) F-NR labeled KB cells in Ca2+-free PBS were unaffected by fs-pulsed irradiation as described above, but immediately produced blebs upon exposure to 0.9 mM Ca2+. For all experiments, cells were incubated with F-NRs for 6 h, then washed 5 times in PBS with 0.9 mM Ca2+ (A-C) or without Ca2+ (D-H). Amounts of dyes and reagents are described as final concentrations in the cell culture medium. Bar = 10 μm.
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