研究目的
The aim of this study was to develop phase-transition nanoparticles (OI_NPs) loaded with perfluoropentane (PFP), indocyanine green (ICG), and oxaliplatin (OXP), to augment anti-tumor efficacy and the immunological effects of chemotherapy, photodynamic therapy and sonodynamic therapy (PSDT).
研究成果
The application of OI_NPs, loaded with appropriate cargo, may represent a novel strategy with which to increase anti-tumor effects, enhance immunological potency, and improve dual-mode imaging.
研究不足
The study only explored the application of OI_NPs plus PSDT in vitro. Future work should investigate this modality in vivo to determine whether the release of tumor antigens into the tumor microenvironment will activate the infiltration of specific immune cells, such as T lymphocyte subsets.
1：Experimental Design and Method Selection:
OI_NPs were fabricated by a double emulsion method and a range of physicochemical and dual-modal imaging features were characterized.
2：Sample Selection and Data Sources:
ID8 cells (syngeneic to C57BL/6 mice) were used.
3：List of Experimental Equipment and Materials:
PLGA, PFP, OXP, ICG, ultrasonic probe, transmission electron microscopy (TEM), scanning electron microscopy (SEM), dynamic light scattering (DLS), ultraviolet-visible (UV-Vis) spectrophotometer, fluorescence spectrometer, confocal laser scanning microscopy (CLSM), flow cytometry, MTT assay, Western blotting, luminometric assays.
4：Experimental Procedures and Operational Workflow:
Cellular uptake of OI_NPs by ID8 cells was determined by CLSM and flow cytometry. The viability and apoptotic rate of ID8 cells were investigated using the MTT assay and flow cytometry. Flow cytometry, Western blotting, and luminometric assays were then used to investigate the exposure or release of crucial DAMPs.
5：Data Analysis Methods:
Data analysis was performed using GraphPad Prism (version 6.0) and all values are shown as mean ± standard deviation (SD). Data were analyzed by analysis of variance (ANOVA) with Dunnett’s test, the unpaired Student’s t-test, or the paired Student’s t-test.
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SEM
FEI Inspect F50
FEI Company
Used to investigate the morphology and structure of the nanoparticles.
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DLS
Zetasizer Nano ZS
Malvern Instruments
Used to determine the diameter, polydispersity index (PDI), and surface potential of different NPs.
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UV-Vis spectrophotometer
260-Bio
Thermo Fisher Scientific
Used to evaluate the entrapment efficiency and loading of ICG.
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fluorescence spectrometer
Cray Eclipse
Agilent Technologies
Used to detect the emission spectra of each sample.
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CLSM
Leica TCS SP8
Leica
Used to evaluate cellular uptake ability.
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TEM
FEI Tecnai G2 F20
FEI Company
Used to investigate the morphology and structure of the nanoparticles.
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PLGA
Medical Polymer Materials
Used as a carrier for the nanoparticles.
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PFP
Strem Chemicals
Used as a component of the phase-transition nanoparticles.
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OXP
MedChemExpress
Used as a chemotherapeutic drug loaded into the nanoparticles.
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ICG
Aladdin
Used as a photo/sonosensitizer loaded into the nanoparticles.
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ultrasonic probe
Sonics & Materials, Inc
Used for sonication in the preparation of nanoparticles.
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ultrasonic diagnostic instrument
MyLab 90
Esaote
Used to capture contrast-enhanced ultrasound (CEUS), and B-mode images.
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PA imaging system
VEVO LASR
FUJIFILM Visual Sonic, Inc
Used to obtain and analyze PA-mode signals of the samples.
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MTT assay
Used to determine cell viability.
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flow cytometry
Used to analyze apoptosis and CRT expression on the cell surface.
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Western blotting
Used to detect the release of HMGB1 into the cell culture supernatant and its translocation to the cytoplasm.
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Luminometric ATP Assay Kit
AAT Bioquest
Used to quantify the secretion of ATP into the supernatant.
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DCFH-DA
Used as an indicator of fluorescence to detect the generation of intracellular ROS.
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lactate cytotoxicity assay kit
Beyotime Co
Used to assess LDH release.
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