JNCI Journal of the National Cancer Institute

Files in this Data Supplement:

• Supplementary Table 1 - Intracellular DOX fluorescence (arbitray units) of tumor cells after DOX delivery in micelles or microbubbles combined or not combined with the application of ultrasound (designated as US). Intracellular DOX fluorescence was calculated as the difference between the measured fluorescence of the tumor cells and autofluorescence of the corresponding control tumor cells.
• Supplementary Fig. 1 - Effect of microbubbles and ultrasound on the intracellular DOX uptake by A2780 ovarian carcinoma cells in suspensions. DOX at a concentration of 0.75 mg/ml was encapsulated in 0.5% PEG-PLLA micelles or 0.5%PEG-PLLA /2% PFP microbubbles; microbubble concentration was 5´108/ml. Continuous wave focused 1.1 MHz ultrasound was applied for 30 s at a peak negative pressure of 2.9 MPa in the focal point (corresponding to a power density of 278 W/cm2). Acoustic droplet evaporation caused microbubble formation and inertial cavitation, which resulted in the enhancement of the intracellular drug uptake.
• Supplementary Fig. 2 - Effect of microbubbles and ultrasound on the intracellular DOX uptake by the cells of the excised MDA MB 231 tumor (sample I of Supplementary Table 1). DOX at a concentration of 0.75 mg/ml was encapsulated in 0.5% PEG-PLLA micelles or 0.5% PEG-PLLA /2% PFP microbubbles; microbubble concentration was 5´108/ml; samples were pre-incubated at 37 ēC for 15 min before ultrasound application; 3-MHz unfocused ultrasound was applied for a 30 s exposure time at a nominal output SATA power density of 2W/cm2 with a duty cycle of 50%.
• Supplementary Fig. 3 - Effect of microbubbles and ultrasound on the intracellular DOX uptake by the MDA MB231 tumors in vivo (sample III) of Table Sm1); 100 ml of DOX formulation was intravenously injected into four MDA MB231 breast cancer tumor bearing mice; a fifth tumor-bearing mouse was used as a control. Two mice were injected with a micellar DOX formulation (0.75 mg/ml DOX /0.5% PEG-PLLA) while two other mice were injected with the microbubble formulation (0.75 mg/ml DOX/0.5% PEG-PLLA/2% PFP) that comprised nanoparticles of two sizes, 250 nm and 750 nm as determined by dynamic light scattering at room temperature. Four hours after the drug injection, the tumor of one mouse in each treatment group was sonicated for 30 s ultrasound exposure time by 3-MHz ultrasound at a nominal output SATA power density of 2 W/cm2 and duty cycle of 20% while the other mouse was not sonicated. Ten minutes after sonication, all mice were sacrificed; tumors and other organs were excised and treated as described in Materials and Methods section to produce individual cells; cell fluorescence was measured by flow cytometry.