Objective: Prepare a multifunctional ultrasound
molecular probe,
hyaluronic acid-mediated
cell-penetrating peptide-modified 10-hydroxycamptothecin-loaded phase-transformation
lipid nanoparticles (HA/CPPs-10-
HCPT-NPs), and to combine HA/CPPs-10-
HCPT-NPs with low-intensity focused ultrasound (LIFU) for precision
theranostics against
hepatocellular carcinoma (HCC). Methods: HA/CPPs-10-
HCPT-NPs were prepared using thin-film dispersion, ultrasound emulsification, and electrostatic effects. HA/CPPs-10-
HCPT-NPs were characterized for particle size, zeta potential, encapsulation efficiency and drug-loading efficiency. In vitro, HA/CPPs-10-
HCPT-NPs were tested for acoustic droplet vaporization (ADV) at different time points/acoustic intensities; the ability of HA/CPPs-10-
HCPT-NPs to target SMMC-7721 cells was detected by confocal
laser scanning microscopy (CLSM); the penetrating ability of CG-TAT-GC-modified NPs was verified by CLSM in a 3D multicellular
tumor spheroid (MCTS) model; the effect of HA/CPPs-10-
HCPT-NPs combined with LIFU on killing SMMC-7721 cells was measured by
CCK-8 and flow cytometry. In vivo, the
tumor-target efficiency of HA/CPPs-10-
HCPT-NPs was evaluated by a small-animal fluorescence imaging system and CLSM; the enhanced ultrasound imaging efficiency of HA/CPPs-10-
HCPT-NPs combined with LIFU was measured by an ultrasound imaging analyzer; the
therapeutic effect of HA/CPPs-10-
HCPT-NPs combined with LIFU was evaluated by
tumor volume,
tumor inhibition rate, and staining (
hematoxylin and
eosin (H & E),
proliferating cell nuclear antigen (
PCNA) and TUNEL). Results: Mean particle size and mean zeta potential of HA/CPPs-10-
HCPT-NPs were 284.2±13.3 nm and - 16.55±1.50 mV, respectively. HA/CPPs-10-
HCPT-NPs could bind to SMMC-7721 cells more readily than CPPs-10-HCPT-NPs. Penetration depth into 3D MCTS of HA/CPPs-10-
HCPT-NPs was 2.76-fold larger than that of NPs without CG-TAT-GC. HA/CPPs-10-
HCPT-NPs could enhance ultrasound imaging by undergoing ADV triggered by LIFU. HA/CPPs-10-
HCPT-NPs+LIFU group demonstrated significantly higher efficiency of anti-proliferation and apoptosis percentage than all other groups. In mouse liver
tumor xenografts, HA/CPPs-10-
HCPT-NPs could target
tumor sites and enhance ultrasound imaging under LIFU. HA/CPPs-10-
HCPT-NPs+LIFU group had a significantly smaller
tumor volume, lower proliferative index (PI), and higher
tumor inhibition and apoptotic index (AI) than all other groups. Conclusions: Combined application of HA/CPPs-10-
HCPT-NPs and LIFU should be a valuable and promising strategy for precise HCC
theranostics.