研究目的
To develop an activatable optoacoustic nanoprobe for noninvasively diagnosing and locating lymphatic metastasis and orthotopic bladder tumor in vivo using multispectral optoacoustic tomography (MSOT) imaging.
研究成果
The activatable optoacoustic nanoprobe successfully enables noninvasive diagnosis and precise localization of lymphatic metastasis and orthotopic bladder tumors in mice via MSOT imaging, demonstrating high specificity for HAase and potential for preclinical tumor monitoring.
研究不足
The study is limited to mouse models, and translation to human applications requires further validation. The sensitivity and specificity in complex biological environments may need optimization. The nanoprobe's performance could be affected by variations in HAase levels and other interfering factors.
1：Experimental Design and Method Selection:
The study designed a nanoprobe based on a tricyanofuran-containing polyene chromophore (TCHM) and hyaluronan (HA) that responds to hyaluronidase (HAase) by aggregation/deaggregation, leading to enhanced optoacoustic signals. MSOT imaging was used for in vivo detection.
2：Sample Selection and Data Sources:
Mouse models were used, including subcutaneous tumor models (4T1 cells), orthotopic bladder tumor models (T24 cells), and lymphatic metastasis models (4T1 cells injected into footpad). Cell lines (4T1, T24, L929) were cultured and characterized.
3：List of Experimental Equipment and Materials:
Equipment includes multispectral optoacoustic tomography system (iThera Medical inVision 128), transmission electron microscope (TEM), dynamic light scattering (DLS), NMR spectrometer, mass spectrometer, UV-Vis spectrophotometer. Materials include synthesized TCHM, modified HA, HAase, phosphate-buffered saline (PBS), DMSO, cell culture reagents, and mice.
4：Experimental Procedures and Operational Workflow:
Synthesis of TCHM and modified HA, preparation of nanoprobe TCHM@HA, in vitro characterization (absorption spectra, OA signals), cytotoxicity assays (MTT), in vivo MSOT imaging of mouse models, histological and biochemical verification (H&E staining, immunofluorescence, Western blotting).
5：Data Analysis Methods:
Spectral unmixing algorithms for MSOT image reconstruction, statistical analysis of OA signal intensities, and biochemical assay data interpretation.
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Multispectral Optoacoustic Tomography System
inVision 128
iThera Medical
Used for in vivo MSOT imaging to detect and visualize optoacoustic signals from the nanoprobe in mouse models.
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Transmission Electron Microscope
Used to characterize the morphology and size of the nanoprobe nanoparticles.
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Dynamic Light Scattering Instrument
Used to measure the hydrodynamic diameter and zeta potential of the nanoprobe.
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NMR Spectrometer
Used for structural characterization of synthesized compounds (e.g., 1H NMR, 13C NMR).
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Mass Spectrometer
Used for mass analysis of synthesized compounds.
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UV-Vis Spectrophotometer
Used to measure absorption spectra and extinction coefficients of the chromophore and nanoprobe.
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Indocyanine Green
ICG
Used as a clinical contrast agent for MSOT imaging to indicate the position of the bladder in control experiments.
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