研究目的
To address the issue of hypoxia attenuating the therapeutic effect of photodynamic therapy (PDT) in cancer treatment by developing a novel strategy using dual-drug nanoparticles to reduce oxygen consumption and enhance PDT efficiency.
研究成果
The dual-drug NPs effectively reduced oxygen consumption, alleviated tumor hypoxia, and enhanced PDT effects, leading to significant tumor growth inhibition and even elimination in mice. This strategy provides a promising approach for improving PDT in hypoxic tumors and could be valuable for clinical applications.
研究不足
The study may have limitations in the scalability of NP production, potential variability in tumor models, and the need for further optimization of drug loading and release kinetics. The in vivo results are specific to mouse models and may not fully translate to human applications without additional studies.
1：Experimental Design and Method Selection:
The study used a film dispersion method to prepare sub-50 nm dual-drug nanoparticles (NPs) encapsulating verteporfin (VER) and atovaquone (ATO) with PLGA-PEG. The design aimed to reduce oxygen consumption and enhance PDT effects in hypoxic tumors. Theoretical models included the enhanced permeability and retention (EPR) effect for tumor targeting.
2：Sample Selection and Data Sources:
Murine breast cancer 4T1 cells were used for in vitro experiments, and 4T1 tumor-bearing Balb/c mice were used for in vivo studies. Data were collected from cell cultures, animal models, and various assays.
3：List of Experimental Equipment and Materials:
Equipment included transmission electron microscopy (TEM), dynamic light scattering (DLS), UV-vis spectroscopy, HPLC, flow cytometer (BD Accuri C6), confocal laser scanning microscopy (CLSM), MTT assay kit, XF96 Analyzer (Seahorse Biosciences), in vivo fluorescence imaging system (Kodak In-Vivo FX Pro), photoacoustic CT scanner (Endra's Nexus 128), and oxygen partial pressure measurement devices (OxyFlopro and oxylitepro). Materials included ATO, VER, PLGA-PEG, DMEM, FBS, SOSG reagent, DCFH-DA probe, and various chemical reagents.
4：Experimental Procedures and Operational Workflow:
NPs were prepared and characterized for size, drug loading, and ROS generation. Cellular uptake, lysosomal escape, oxygen consumption, ROS production, cytotoxicity, and in vivo biodistribution and antitumor efficacy were assessed step by step with specific protocols for each assay, including laser irradiation for PDT effects.
5：Data Analysis Methods:
Data were analyzed using statistical methods such as Student's t-test and one-way ANOVA, with software tools for flow cytometry, microscopy, and imaging systems.
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High Performance Liquid Chromatography System
Shimadzu LC-20A
Shimadzu
Used for drug content analysis and quantification.
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Confocal Laser Scanning Microscope
ZEISS CLSM 700
ZEISS
Used for intracellular localization and lysosome escape studies.
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Transmission Electron Microscope
Not specified
Not specified
Used for imaging nanoparticles to characterize their size and morphology.
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Dynamic Light Scattering Instrument
Not specified
Not specified
Used to measure the hydrodynamic size and polydispersity index of nanoparticles.
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UV-Vis Spectrophotometer
Not specified
Not specified
Used to measure absorption spectra of drugs and nanoparticles.
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Flow Cytometer
BD Accuri C6
BD
Used for analyzing cellular uptake and ROS production via fluorescence intensity.
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MTT Assay Kit
Not specified
Not specified
Used for cell viability and cytotoxicity assessments.
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XF96 Analyzer
XF96
Seahorse Biosciences
Used to measure oxygen consumption rate in cells.
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In Vivo Fluorescence Imaging System
Kodak In-Vivo FX Pro
Kodak
Used for biodistribution and tumor imaging in mice.
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Photoacoustic CT Scanner
Endra's Nexus 128
Endra
Used for in vivo photoacoustic imaging to assess tumor oxygenation.
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Oxygen Partial Pressure Measurement Device
OxyFlopro and oxylitepro
Oxford Optronix
Used to directly measure oxygen partial pressure in tumor tissues.
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Laser
635 nm laser
Not specified
Used for irradiating samples to induce photodynamic therapy effects.
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