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Purpose  Tuberculosis (TB) is a highly infectious disease caused by  Mycobacterium tuberculosis  ( Mtb ), which often parasites in macrophages. This study is performed to investigate the bactericidal effect and underlying mechanisms of low-frequency and low-intensity ultrasound (LFLIU) combined with levofloxacin-loaded PLGA nanoparticles (LEV-NPs) on  M. smegmatis  (a surrogate of  Mtb ) in macrophages.    Methods and results  The LEV-NPs were prepared using a double emulsification method. The average diameter, zeta potential, polydispersity index, morphology, and drug release efficiency in vitro of the LEV-NPs were investigated.  M. smegmatis  in macrophages was treated using the LEV-NPs combined with 42 kHz ultrasound irradiation at an intensity of 0.13 W/cm 2  for 10 min. The results showed that ultrasound significantly promoted the phagocytosis of nanoparticles by macrophages ( P  < 0.05). In addition, further ultrasound combined with the LEV-NPs promoted the production of reactive oxygen species (ROS) in macrophage, and the apoptosis rate of the macrophages was significantly higher than that of the control ( P  < 0.05). The transmission electronic microscope showed that the cell wall of  M. smegmatis  was ruptured, the cell structure was incomplete, and the bacteria received severe damage in the ultrasound combined with the LEV-NPs group. Activity assays showed that ultrasound combined with the LEV-NPs exhibited a tenfold higher antibacterial activity against  M. smegmatis  residing inside macrophages compared with the free drug.    Conclusion  These data demonstrated that ultrasound combined with LEV-NPs has great potential as a therapeutic agent for TB.

journal of nanobiotechnology

A synergistic bactericidal effect of low-frequency and low-intensity ultrasound combined with levofloxacin-loaded PLGA nanoparticles on M. smegmatis in macrophages