研究目的
Investigating the construction and application of a neutrophil-based living probe for precision tumor imaging diagnosis.
研究成果
The neutrophil-based living probe demonstrated faster targeting and stronger accumulation in tumor sites compared to nanoparticle-based imaging agents, indicating its great potential for precision tumor-diagnosis applications. The study highlights the importance of leveraging natural biological properties of cells for enhanced imaging and diagnostic capabilities.
研究不足
The study is limited by the potential variability in neutrophil collection and the need for further optimization of nanoparticle conjugation to ensure minimal impact on neutrophil function. Additionally, the translation of these findings to clinical settings requires further investigation.
1:Experimental Design and Method Selection:
The study utilized a cellular-surface engineering approach to construct a neutrophil-based living probe for tumor imaging. Imaging agents (Gd and BODIPY) loaded BSA nanoparticles were covalently fixed onto the cellular surface of neutrophils through a TNB-mediated disulfide–thiol exchange.
2:Sample Selection and Data Sources:
Murine neutrophils were collected from the peritoneal cavity of BALB/c mice. The purity and functionality of these neutrophils were analyzed before and after modification with nanoparticles.
3:List of Experimental Equipment and Materials:
Included bovine serum albumin (BSA), gadolinium (Gd), BODIPY, 5-thio-2-nitrobenzoate (TNB), and various antibodies for flow cytometry and confocal microscopy.
4:Experimental Procedures and Operational Workflow:
Neutrophils were incubated with DTNB-activated BSA nanoparticles. The biological properties of the resultant neutrophil probe were investigated, including cell viability, morphology, and chemotaxis in vivo. The tumor-targeting efficacy was evaluated in a mouse lung-cancer model using fluorescence and MR imaging.
5:Data Analysis Methods:
Flow cytometry, confocal laser-scanning microscopy, and MR imaging were used to analyze the data. Statistical analysis was performed using one- or two-way ANOVA.
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bovine serum albumin
BSA
Sigma-Aldrich
Used as a scaffold for loading imaging agents (Gd and BODIPY) onto nanoparticles.
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BODIPY 650/665-X N-hydroxysuccinimide ester
BODIPY 650/665-X NHS ester
Life Technologies
Used as an imaging agent loaded onto BSA nanoparticles.
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FITC-conjugated CD11b antibody
eBiosciences
Used for flow cytometry analysis to evaluate the expression of CD11b on neutrophils.
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R-phycoerythrin (PE)/Cy7-conjugated Ly-6G antibody
BioLegend
Used for flow cytometry analysis to evaluate the purity of neutrophils.
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Annexin V-FITC apoptosis detection kit
Beyotime
Used to detect the viability of nanoparticle-remodeled neutrophils.
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cell counting kit-8
CCK-8
Beyotime
Used to estimate the in vitro cytotoxicity of Gd@BSA NPs.
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Giemsa stain kit
Nanjing Jiancheng Bioengineering Institute
Used to stain neutrophils for morphological observation.
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red blood cell lysis buffer
RBC lysis buffer
Peprotech-BioGems
Used to lyse red blood cells during neutrophil collection.
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