Application of nanoparticle loaded with indocyanine green and adriamycin in in vitro imaging of retinoblastoma： An experimental study

Abstract:

Objective：To prepare the phase-change multifunctional lipid nanoparticle containing indocyanine green（ICG） and adri－amycin（ADM） and targeting folate receptor（ICG/FA/Pct-PFPNP：ADM），to investigate its basic properties，targeting ability，pho－tothermal effect，and ultrasonic/photoacoustic bimodal imaging ability，and to provide new strategies for the diagnosis of retinoblas－toma. Methods：ICG/FA/Pct-PFPNP：ADM nanoparticles were prepared by the double emulsification method. A transmission electron microscope was used to observe the morphology of nanoparticles，the Malvern particle size analyzer was used to measure particle size and electric potential，and an ultraviolet spectrophotometer was used to measure the encapsulation efficiency and drug loading rate of nanoparticles. Flow cytometry and confocal laser scanning were used to observe the expression of folic acid on the surface of ICG/FA/Pct-PFPNP：ADM nanoparticles and targeting ability in vitro. CCK-8 assay was used to evaluate the biosafety of nanoparticles. Near-infrared laser（808 nm） was used to excite ICG/FA/Pct-PFPNP：ADM nanoparticles and assess their ultrasonic/photoacoustic bimodal imaging ability. Results：ICG/FA/Pct-PFPNP：ADM nanoparticles were successfully prepared，with a spherical appearance，an average particle size of （374.40±23.48） nm，and an average potential of （-28.0±4.5） mV. The encapsulation efficiency and drug load－ing rate of ICG were （91.85±2.98）% and （9.19±0.29）%，respectively，and the encapsulation efficiency and drug loading rate of ADM were （62.04±5.10）% and （6.20±0.51）%，respec－tively. Both flow cytometry and laser confocal microscopy showed good expression of folic acid on the surface of the nanoparticles；the targeted group had a Y79 cell connection rate of （96.57±3.17）%，which was significantly different from that in the non-targeted group and the antibody blocking group（F=1537.6，P<0.05）. The CCK-8 assay showed no obvious cytotoxicity of ICG/FA/Pct-PFPNP：ADM nanoparticles. The ICG/FA/Pct-PFPNP：ADM nanoparticles showed liquid-gas phase transition after laser irradiation，enhanced ultrasound imaging，and increased local temperature（60.8 ℃）. The intensity of photoacoustic signal increased with the increase in nanoparticle concentration，and there was a good linear relationship between nanoparticle concentration and signal intensity. Conclusion：The phase-change nanoparticles loaded with ICG and ADM were successfully prepared，with good targeting ability and photothermal effect. Such nanoparticles can enhance the quality of ultrasonic/photoacoustic imaging. Therefore，it can be used in the diagnosis of retinoblastoma.