Basic Research On DVDMS/IR780-load Composite Nanoparticles In Dual-mode Imaging And Sonodynamic Treatment Of Cancer Cell

Objective: To synthesize the composite nanoparticles by encapsulating sinoporphyrin sodium(DVDMS),IR780 and Perfluoropentane(PFP).Testify their physicochemical properties and biosafety.Methods: DVDMS@IR780@PFP@PLGA(DIPP-NPs)were synthesized via a double emulsion method.IR780 was loaded on the shell,DVDMS and PFP were loaded into the core.Simultaneously,PFP@PLGA(PP-NPs),DVDMS@PFP@PLGA(DPP-NPs)and IR780@PFP@PLGA(IPP-NPs)were also synthesized by the same method.The apparent differences of various nanoparticles were observed by the digital photographs.The morphology and structure of the DIPP-NPs were characterized by optical microscopy and transmission electron microscopy(TEM).The average size,zeta potential and the stability of the DIPP-NPs were analyzed were determined by Malvern Instruments.UV-vis spectrophotometer was used to determine the UV-vis absorption spectrum of Free-DVDMS,Free-IR780,PP-NPs and DIPP-NPs.The encapsulation efficiency of DVDMS and IR780 were calculated by corresponding standard curve.The cytotoxicity of DIPP-NPs at various concentrations(20,40,60,80,100 ug/ml)was detected by CCK-8.Kunming mice were randomly divided into 5 groups(n=3;control,3,7,14 and 30 day groups)for blood routine analysis,blood biochemical analysis and H&E staining of main organs.Results: DVDMS@IR780@PFP@PLGA(DIPP-NPs)were successfully synthesized,appeared as dark green emulsion with good dispersion and appeared as spheres with homogeneous size as revealed by optical microscopy and TEM.The average size was(222.39 ± 8.82)nm and no obvious changes within 7 days,the zeta potential was(-19.43 ± 0.41)mv.DIPP-NPs in DMSO exhibited characteristic peaks at 400 nm(DVDMS)and 793 nm(IR780).The loading efficiencies of IR780 and DVDMS were(94.84 ± 3.09)% and(90.85 ± 6.55)%,respectively.In addition,DIPP-NPs showed a good biosafety.Conclusion: The composite nanoparticles DIPP-NPs were successfully synthesized,which has stable structure and high biological safety and can be used in further experiments.Objective: To investigate the cellular uptake and the ROS generation of DIPP-NPs in 4T1 cells,explore the synergistic enhancement of SDT between DVDMS and IR780.Methods: The intracellular uptake of PP-NPs,DPP-NPs,IPP-NPs and DIPP-NPs were observed by confocal laser scanning microscopy(CLSM)and flow cytometry(FCM).The ROS generation of DPP-NPs,IPP-NPs and DIPP-NPs were evaluated by SOSG and DCFH-DA.The sonotoxicity against 4T1 cells was evaluated by CCK-8 assay,CLSM and FCM in vitro.The biological distribution of DIPP-NPS was evaluated by fluorescence imaging.The therapeutic effect was evaluated by tumor inhibition rate in vivo via monitoring the changes of tumor volume/weight.Results: DIPP-NPs have a good performance of tumor targeting in vitro.AS the CLSM images shown,numerous DIPP-NPs aggregated around the 4T1 cells and the cellular uptake efficiency was(97.21 ± 1.26)%.Besides,the ROS generation performance of DIPP-NPs was verified and the apoptosis rate reached(57.43 ± 3.26)% in vitro.In addition,DIPP-NPs have a good performance of tumor targeting in vivo.The tumor volume and weight inhibition rate in DPP-NPs + US of 4T1 tumor-bearing mice increased approximately(82.70 ± 4.35)% and(81.69 ± 1.61)%,respectively.The tumor volume and weight inhibition rate in DPP-NPs + US of CT26 tumor-bearing mice increased approximately(81.51 ± 4.58)% and(83.34 ± 5.10)%,respectively.Conclusion: DIPP-NPs have excellent tumor targeting and reactive oxygen generation capacities.Most importantly,DIPP-NPs have obvious anti-tumor effect,whether it’s superficial breast cancer or deep colon cancer.This study provides treatments of tumors with more possibilities.Objective: To explore the acoustic droplet vaporization(ADV)and PA/US dual-mode imaging abilities of DIPP-NPs both in vitro and in vivo.Methods:(1)ADV and US imaging: LIFU,a US transducer,was used to irradiate the DIPP-NPs emulsion(1 mg/m L)at the intensity of 2.4 W/cm2 for different durations(0,60,120,180,240,300,360,420s).After US irradiation,the optical microscopy images were recorded,US images and corresponding echo intensities were performed with the US system.Established female tumor-bearing mouse models and DIPP-NPs(3 mg/ml,200 ul)was injected through the tail vein.Then,US(2.4 W/cm2,180s)was used to stimulate the tumor tissue at 8 h after injection.Finally,US images and corresponding echo intensities were performed with the US system mentioned above.(2)PA imaging: DIPP-NPs emulsion(1mg/ml)was scanned within 680~970 nm by Photoacoustic Imaging System to detect the optimal excitation wavelength.Then,PA images of the DIPP-NPs emulsion at different concentrations(0.5,1.0,1.5,2.0,2.5mg/m L)and corresponding PA signal intensities were analyzed.Established female tumor-bearing mouse models and DIPP-NPs(3 mg/ml,200 ul)was injected through the tail vein.The PA images in the tumor regions were acquired at different time points(0,2,4,6,and 8 h)after injection and the PA intensities were analyzed at corresponding time.Results: Substantial amounts of DIPP-NPs were converted into MBs after US irradiation was observed by light microscope.The signal intensities of DIPP-NPs increase with the increase of irradiation time in vitro and the signal intensity reached the highest after US irradiation in vivo.The PA signal intensity of DIPP-NPs increase with the increase of concentrations and the signal intensity reached the highest at 8 h after injection.Conclusion: DIPP-NPs showed great PA/US imaging abilities,which led to diagnostic-imaging guidance and monitoring during tumor therapy.