Figure 9. Nanoparticle Uptake in OVCAR5-WT and Hypoxic Cells.

Panel A: Nanoparticle formulations were loaded with rhodamine 123 (green). OVCAR5 wild type (WT) and hypoxic (HYP) cells were treated with targeted nanoparticles (T), targeted nanoparticles in the presence of excess competitive EGFR antibody (TC), non-targeted nanoparticles (N), and non-targeted nanoparticles in the presence of excess competitive EGFR antibody (NC). Cells were imaged after 15 minutes and 30 minutes of treatment (A). Cell nuclei were stained with Hoechst 33342 (blue). The fluorescent images represent a merge of the blue and green fluorescence; corresponding DIC images are also shown. Targeted nanoparticle uptake is apparent at the early time points and is blocked by the competitive antibody. Panel B: Cells were treated with the rhodamine 123 loaded formulations, and the amount of rhodamine 123 taken up by the cells was quantified and converted to a percentage of the administered dose. Normoxic treatments are shown in black while the hypoxic treatments are shown in blue. Cells were treated with the following nanoparticle formulations; non-targeted nanoparticles (solid white bar for normoxic cells; solid blue bar for hypoxic cells), targeted nanoparticles (solid black bar for normoxic cells; blue checkered bar for hypoxic cells), and targeted nanoparticles in the presence of excess competitive EGFR antibody (black/white speckled bar for normoxic cells; blue diagonal line bar for hypoxic cells). Cells were treated for 15 minutes, 30 minutes, 1 hour, 2 hours, 3 hours, and 6 hours. Fluorescence was quantified using a Bio-Tek Synergy HT plate reader. Each treatment represents an n=8.