研究目的
To develop NGO@DOX-PEG nanodrugs for dual-responsive photochemical therapy to improve biocompatibility and therapeutic effects in antitumor applications.
研究成果
NGO@DOX-PEG nanodrugs, particularly NGO@DOX-PEG5K, exhibit excellent biocompatibility, pH and NIR dual-responsive drug release, enhanced cytotoxicity, cellular uptake, and nuclear translocation upon NIR irradiation, making them promising for photochemical antitumor therapy. The noncovalent modification approach effectively improves stability and therapeutic efficacy.
研究不足
The study is limited to in vitro experiments using HeLa cells; in vivo studies and long-term biocompatibility assessments are not included. The noncovalent modification may lead to variability in drug loading and release, and the specific mechanisms of NIR-enhanced effects could require further investigation.
1：Experimental Design and Method Selection:
The study involved synthesizing DOX-PEG polymers with different molecular weights (PEG2K, PEG5K, PEG20K) and noncovalently modifying nano-graphene oxide (NGO) to create NGO@DOX-PEG nanodrugs. Methods included Hummer's method for NGO preparation, amide bond conjugation for DOX-PEG synthesis, and dialysis for purification.
2：Sample Selection and Data Sources:
HeLa cells were used for in vitro experiments, sourced from cell culture. Materials included DOX, NGO, PEG-COOH, and various reagents from specified suppliers.
3：List of Experimental Equipment and Materials:
Equipment included Transmission Electron Microscope (JEOL), Zetasizer Nano S90 (Malvern), UV-Vis spectrometer, FTIR spectrometer, fluorescence spectrometer, confocal laser scanning microscope (Leica), and NIR laser (808 nm). Materials included DOX from Macklin Inc., multilayer graphene oxide from Hengqiu technology, PEG-COOH from Shanghai Ponsure Biotech, and inhibitors like chlorpromazine and amiloride from aladdin Reagent Inc.
4：Experimental Procedures and Operational Workflow:
NGO was prepared and characterized; DOX-PEG was synthesized and loaded onto NGO via sonication and stirring; nanodrugs were characterized using UV-Vis, FTIR, TEM, DLS; photothermal effects were tested with NIR irradiation; drug release was studied at different pH with/without NIR; cytotoxicity was assessed via MTT assay; ROS generation, cellular uptake, endocytosis pathways, and intracellular distribution were evaluated using fluorescence-based methods and inhibitors.
5：Data Analysis Methods:
Data were analyzed using one-way ANOVA for statistical significance, with results expressed as mean ± standard deviation (SD).
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Zetasizer Nano S90
S90
Malvern
Measuring zeta potentials and mean particle sizes of nanodrugs
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Transmission Electron Microscope
JEOL
Observing the morphologies of NGO and NGO@DOX-PEG nanodrugs
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Confocal Laser Scanning Microscope
Leica
Observing intracellular distribution of nanodrugs
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NIR Laser
808 nm
Irradiating samples for photothermal and drug release studies
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UV-Vis Spectrometer
Detecting drug loading and release characteristics
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FTIR Spectrometer
Analyzing chemical bonds and functional groups in nanodrugs
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Fluorescence Spectrometer
Measuring fluorescence intensity for drug release and ROS detection
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Microplate Reader
Testing absorbance in MTT assays for cell viability
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Doxorubicin
Macklin Inc.
Anticancer drug used in nanodrug synthesis
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Multilayer Graphene Oxide
Hengqiu technology
Starting material for preparing NGO
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PEG-COOH
PEG2K-COOH, PEG5K-COOH, PEG20K-COOH
Shanghai Ponsure Biotech, Inc
Polymer for conjugating with DOX to form DOX-PEG
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EDCI
aladdin Reagent Inc
Reagent for amide bond formation in DOX-PEG synthesis
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NHS
aladdin Reagent Inc
Reagent for amide bond formation in DOX-PEG synthesis
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DMSO-d6
aladdin Reagent Inc
Solvent for NMR analysis
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Chlorpromazine
CPZ
aladdin Reagent Inc
Endocytosis inhibitor for studying cellular uptake pathways
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Amiloride
EIPA
aladdin Reagent Inc
Endocytosis inhibitor for studying cellular uptake pathways
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Methyl-betacyclodextrin
M-β-CD
aladdin Reagent Inc
Endocytosis inhibitor for studying cellular uptake pathways
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MTT
aladdin Reagent Inc
Reagent for cell viability assays
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Dichlorofluorescein diacetate
DCFH-DA
Wanlei Biotechnology
Fluorescent probe for detecting intracellular ROS levels
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Hoechest 33342
Wanlei Biotechnology
Nuclear stain for confocal microscopy
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