Preparation Of Nanomicelles Co-loaded With An Aggregation-induced Emission Dye And Gold Nanoparticles And Study Of Their In Vivo Tumor-targeted Dual-modal Fluorescence/CT Imaging Effects

ObjectiveTo prepare the nanomicelles co-loaded with an aggregation-induced emission dye and gold nanoparticles,and study their in vivo tumor-targeted dual-modal fluorescence/CT imaging abilities.Methods1.Preparation,characterization and toxicity studies of M-NPAPF-Au.The nanomicelles of M-NPAPF-Au were facile fabricated via “one-pot ultrasonic emulsification”.Particle size was determined by dynamic light scattering(DLS)using a particle size determination instrument.The toxicity of M-NPAPF-Au was evaluated using three kinds of cells: CT26,Hep G2 and L02.2.In vitro fluorescence/CT imaging effects of M-NPAPF-Au.Studying the AIE properties of M-NPAPF-Au;Using flow cytometric analysis,confocal laser scanning microscopy(CLSM)and spectrum imaging system to analy the in vitro fluorescence imaging effects of M-NPAPF-Au;Using a CT scanning system to detect the in vitro CT imaging signal of M-NPAPF-Au.The laser excitation of spectrum imaging system was 488 nm and the collecting wave lengths were from 660 nm to 750 nm;The measurement parameters of CT scanning system were set as follows: effective pixel size,50 ?m;80 k Vp,500 ?A;field of view,91.07 mm × 91.07 mm × 91.07 mm;fly,360.3.In vivo tumor-targeted dual-modal fluorescence/CT imaging abilities of M-NPAPF-Au.In vivo fluorescence imaging,biodistribution and semi-quantitative phar-macokinetics studies;In vivo CT imaging and biodistribution.The animal model was tumor-bearing(CT26 cells)female BALB/c mice.Results1.Fabricating M-NPAPF-Au by co-loading the AIE red dye(NPAPF)and gold nanoparticles into the well-known FDA-approved material 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethyleneglycol)-2000] DS PE-PEG2 0 0 0 micelles.The 1:1:8 ratio of Au/NPAPF/DSPE-PEG2 0 0 0(w/w/w)was chosen to prepare the nanomicelles.The particle size of M-NPAPF-Au was ~ 65 nm.DLS studies showed that the micelles had good mono-dispersity.And the particle size was particularly favorable for tumor-targeted in vivo imaging due to the EPR effect.And the study of cells treated by M-NPAPF-Au within concentration of 600 ?g/ml and mice within 7 mg/ml showed the toxicity of M-NPAPF-Au was inapparent simultaneously,indicating that the as-prepared M-NPAPF-Au had superior biocompatibility.2.The results of in vitro fluorescence/CT imaging effects of M-NPAPF-Au indicated that NPAPF retained the crucial AIE feature in the as-prepared nanomicelles system despite the existence of gold nanoparticles,which guaranteed its efficient fluorescence imaging capacity.M-NPAPF-Au had excellent fluorescence emission properties and CT imaging signal,this is an important discovery in the field of fluorescence/CT imaging agent.3.The studies of in vivo tumor-targeted dual-modal fluorescence/CT imaging demonstrated that M-NPAPF-Au had good biocompatibility,long blood circulation half-life,superior tumor-targeting ability and excellent fluoresceence/CT imaging effects.And the studies assuredly indicated the significant potential application of M-NPAPF-Au for in vivo tumor-targeted dual-modal fluorescence/CT imaging and diagnosis.ConclusionsDeveloped a promising preparation method of nanomicelles(M-NPAPF-Au)co-loaded with NPAPF and gold nanoparticles.M-NPAPF-Au has excellent in vivo tumor-targeted dual-modal fluorescence/CT imaging abilities.