研究目的
To develop a novel phototheranostic nanocomplex self-assembled from bovine serum albumin (BSA) and indocyanine green (ICG) for enhanced near-infrared (NIR) fluorescence imaging to guide in vivo cancer photothermal therapy (PTT).
研究成果
The ICG–BSA NC is a promising phototheranostic nanoplatform for highly sensitive NIR fluorescence imaging and imaging-synergized cancer PTT treatment, demonstrating efficient tumor accumulation, improved hydrolytic stability, and significant tumor growth suppression without notable side effects.
研究不足
The study focuses on the optimization of ICG loading ratio and its effects on PLQY and hydrolytic stability. The potential long-term effects and clinical translation of the ICG–BSA NC require further investigation.
1：Experimental Design and Method Selection:
The nanocomplex was prepared by the self-assembling of BSA and ICG via nanoprecipitation method. The binding of ICG with the binding sites on the albumin was confirmed to result in improved hydrolytic stability and high photoluminescence quantum yield (PLQY).
2：Sample Selection and Data Sources:
Triple negative breast cancer 4T1 cells and six-week old female Bal b/c mice were used for in vitro and in vivo studies, respectively.
3：List of Experimental Equipment and Materials:
ICG, BSA, ethanol (anhydrous), dimethyl sulfoxide (DMSO), 4′,6-diamidino-2-phenylindole (DAPI), RPMI 1640 cell culture medium, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), fetal bovine serum (FBS), trypsin–EDTA solution, penicillin–streptomycin solution, and 1× PBS buffer (pH 7.4).
4：4). Experimental Procedures and Operational Workflow:
4. Experimental Procedures and Operational Workflow: The ICG–BSA NC was characterized by SEM, TEM, DLS, UV–Vis–NIR absorption spectra, and fluorescence emission spectra. The PLQY was determined, and the hydrolytic stability was investigated. In vitro cytotoxicity, cellular uptake, and NIR FI were evaluated. In vivo NIR FI, biodistribution, and PTT effects were assessed.
5：Data Analysis Methods:
The fluorescence signals were extracted and analyzed using a Maestro? in vivo fluorescence imaging system. The tumor volumes and body weights were measured, and histological examinations were performed.
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Confocal laser scanning microscope
Leica, TCS-SP5
Leica
Used for cellular uptake and NIR fluorescence imaging in vitro.
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Indocyanine green
Sigma-Aldrich LLC
Used as a fluorescent dye for NIR fluorescence imaging and photothermal therapy.
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Bovine serum albumin
Sigma-Aldrich LLC
Used as a biocompatible carrier material for the nanocomplex.
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Ethanol
anhydrous
Sigma-Aldrich LLC
Used in the preparation of the nanocomplex to reduce albumin solubility.
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Dimethyl sulfoxide
Sigma-Aldrich LLC
Used as a solvent in the preparation of the nanocomplex.
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4′,6-diamidino-2-phenylindole
DAPI
Sangon Biotech
Used for staining cells in confocal laser scanning microscope imaging.
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RPMI 1640 cell culture medium
Sangon Biotech
Used for culturing 4T1 cells.
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3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
MTT
Sangon Biotech
Used for evaluating cell viability.
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Fetal bovine serum
FBS
Sangon Biotech
Used as a supplement in cell culture medium.
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Trypsin–EDTA solution
Sangon Biotech
Used for detaching cells from culture dishes.
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Penicillin–streptomycin solution
Sangon Biotech
Used as an antibiotic in cell culture medium.
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PBS buffer
1×, pH 7.4
Sangon Biotech
Used for washing cells and diluting reagents.
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Maestro? in vivo fluorescence imaging system
CRI Inc.
Used for in vivo fluorescence imaging.
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LAMBDA 750 UV/Vis/NIR spectrophotometer
Perkin Elmer
Used for obtaining absorption spectra.
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FluoroMax 4
Horiba Jobin–Yvon
Used for measuring fluorescence emission spectra.
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Zetasizer Nano-ZS90
Malvern
Used for characterizing dynamic diameter and distribution of nanoparticles.
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FEI Quanta 200F SEM
FEI
Used for SEM characterization of the nanocomplex.
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FEI Tecnai G2 F20 S-Twin TEM
FEI
Used for TEM characterization of the nanocomplex.
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