Mechanism of the ultrasonic activation of micellar drug delivery

Abstract

The mechanism of the ultrasonic enhancement of the uptake of cytotoxic drugs, doxorubicin (DOX) and ruboxyl (Rb) by HL-60 cells from Pluronic micelles was studied. DOX and Rb sorption from either PBS or micellar Pluronic solutions is described by Langmuir-type isotherms characteristic of substrates with limited number of sorption centers. The sorption limits for Rb from PBS and Pluronic were considerably higher than those for DOX, presumably due to much higher Rb partitioning into cell membranes. The overall number of drug sorption centers for both drugs decreased in the presence of Pluronic implying the effect of Pluronic on the DNA conformation, which was confirmed by the electron paramagnetic resonance (EPR) experiments using Rb as a spin probe. Ultrasound increased drug uptake by the cells from PBS and Pluronic solutions. The fluorescence microscopy and flow cytometry experiments using fluorescently-labeled Pluronic showed that ultrasound enhanced both the intracellular uptake of Pluronic micelles and Pluronic trafficking into cell nuclei. A scheme is suggested that describes various equilibria controlling drug/cell interactions and effect of ultrasound on these equilibria. Under the action of ultrasound, the equilibrium between the micellar-encapsulated and free drug is shifted in the direction of free drug due to micelle perturbation; the equilibrium between extracellular and internalized drug is shifted to the intracellular drug due to the ultrasound-induced cellular changes that enhance the accessibility of various cellular structures to drug. An important advantage offered by ultrasound is that the same degree of the intracellular drug uptake may be achieved at a substantially lower drug concentration in the incubation medium.