研究目的
Investigating the therapeutic effects of a carboxyl-terminated dendrimer encapsulated with platinum nanoparticles for targeted delivery and photothermal ablation of malignant bone tumors.
研究成果
The carboxyl-terminated dendrimer was developed as a novel vehicle to favorably deliver Pt nanoparticles to the osteolytic bone lesions for PTT of bone tumors. The plentiful carboxyl groups on the dendrimer surface endowed DEPt-COOH the intrinsic bone-binding affinity. DEPt-COOH represented high affinity to hydroxyapatite and bone fragments in vitro. The biodistribution revealed that DEPt-COOH were able to selectively recognize the osteolytic bone lesions in bone tumors, resulting in a higher accumulation in the tumor-bearing tibias than in the healthy ones. The therapeutic experiments demonstrated that DEPt-COOH-mediated PTT efficiently regressed the bone tumor growth and tumor-associated osteolysis.
研究不足
The study focuses on a specific model of malignant bone tumors and the use of carboxyl-terminated dendrimers for targeted delivery. The generalizability to other types of tumors or delivery systems is not explored. The long-term effects and potential side effects of the treatment are not fully investigated.
1:Experimental Design and Method Selection:
The study synthesized carboxyl-terminated dendrimer-encapsulated platinum (Pt) nanoparticles (DEPt-COOH) for targeted photothermal therapy (PTT) of malignant bone tumors. Amine-terminated dendrimer-encapsulated Pt nanoparticles (DEPt-NH2) and acetylated DEPt-NH2 (DEPt-AC) were synthesized as control materials. The bone-binding affinity of the three materials was assessed on hydroxyapatite or bone in vitro and in vivo.
2:Sample Selection and Data Sources:
NIH3T3 cells and MDA-MB-231 cells stably expressing luciferase (MDA-MB-231-Luc cells) were used.
3:List of Experimental Equipment and Materials:
G5-NH2 PAMAM dendrimer, G
4:5-COOH PAMAM dendrimer, acetic anhydride, triethylamine, potassium hexachloroplatinate (K2PtCl6), sodium borohydride (NaBH4), hydrochloric acid, methanol, sodium hydroxide. Experimental Procedures and Operational Workflow:
Synthesis of DEPts, characterization, photothermal effect assessment, bone-binding affinity assay, cytotoxicity assay, in vitro killing cancer cells by DEPt-associated PTT, in vivo biodistribution, in vivo treatment of malignant bone tumors by PTT, tumor apoptosis assay, histopathological examination, tumor-bearing tibia reconstruction by micro-CT, hemolysis assay, blood test assay, excretion analysis.
5:Data Analysis Methods:
Statistical significance was analyzed by student's t-test.
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Transmission electron microscopy
HT7700
Hitachi
Used for imaging the synthesized nanoparticles.
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High-resolution TEM
JEM-2100
JEOL
Used for high-resolution imaging of nanoparticles.
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Zetasizer Nano
ZS90
Malvern Instruments
Used for determining the hydrodynamic sizes of nanoparticles.
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MC-ICP-MS
Neptune
Thermo
Used for quantitative analysis of Pt element in samples.
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UV-Vis spectrophotometer
Cary 60
Agilent Technologies
Used for recording the ultraviolet-visible absorption spectra of nanoparticles.
Cary 60 UV-Vis Spectrophotometer
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G5-NH2 PAMAM dendrimer
Dendritech Inc.
Used as a scaffold for the synthesis of dendrimer-encapsulated nanoparticles.
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G4.5-COOH PAMAM dendrimer
Dendritech Inc.
Used as a scaffold for the synthesis of dendrimer-encapsulated nanoparticles.
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Potassium hexachloroplatinate
Sigma-Aldrich
Used in the synthesis of platinum nanoparticles.
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Sodium borohydride
Sigma-Aldrich
Used as a reducing agent in the synthesis of platinum nanoparticles.
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NIR laser
CNI laser
Used for photothermal therapy experiments.
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Infrared camera
Magnity Electronics
Used for recording temperature changes during photothermal therapy.
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IVIS Lumina II in vivo imaging system
Caliper Life Sciences
Used for recording the luminescence images of mice.
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Micro-CT scanner
Skyscan 1077
Used for scanning and reconstructing the 3D features of tumor-bearing tibias.
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