研究目的
To develop a near-infrared (NIR)-modulated thermosensitive hydrogel (PNT-gel) for stabilizing indocyanine green (ICG) and enabling efficient combinatorial photothermal/photodynamic antitumor therapy, addressing the limitations of ICG such as instability, short plasma half-life, and lack of targeting ability.
研究成果
The PNT-gel successfully stabilized ICG, enabled controlled release via NIR irradiation, and achieved efficient combinatorial photothermal/photodynamic therapy, leading to almost complete tumor eradication without recurrence in mouse models. This multifunctional hydrogel platform offers a promising strategy for localized, repeated cancer therapy with minimal side effects.
研究不足
The study focuses on a specific hydrogel system and ICG; generalizability to other phototherapeutic agents or cancer types may be limited. In vivo degradation and long-term effects beyond 14 days were not extensively studied. The use of low-dose NIR laser may not be sufficient for all tumor types, and potential immune responses or side effects in humans were not addressed.
1：Experimental Design and Method Selection:
The study designed a supramolecular cross-linking conjugated polymer-based hydrogel (PNT-gel) using host-guest assembly between α-cyclodextrin (α-CD) and PEGylated poly(N-phenylglycine) (PPG-g-peg) to form a photothermal network. Methods included oxidative polymerization, esterification, and gelation via ultrasonication and incubation.
2：Sample Selection and Data Sources:
Samples included synthesized PPG-g-peg, PNT-gel, and ICG-loaded PNT-gel. Cell lines used were L929 mouse fibroblast and 4T1 mouse breast cancer cells. Animal models were BALB/c nude mice and Kunming mice for in vivo studies.
3：List of Experimental Equipment and Materials:
Equipment included FTIR spectrometer, NMR spectrometer, TGA system, UV-vis-NIR spectrophotometer, XRD diffractometer, rheometer, infrared thermal camera, SEM, CLSM, flow cytometer, and NIR laser. Materials included N-phenylglycine, PEG, α-CD, ICG, and various chemicals from suppliers like Aladdin Chemistry Co., Ltd., and Sigma-Aldrich.
4：Experimental Procedures and Operational Workflow:
Procedures involved synthesis of PPG-g-peg, preparation of PNT-gel, characterization (FTIR, NMR, TGA, UV-vis-NIR, XRD, rheology), photothermal performance tests, in vitro ICG release studies, cytotoxicity assays, cellular uptake and ROS detection, in vitro phototherapy, and in vivo animal experiments including gelation, retention, imaging, and therapy.
5：Data Analysis Methods:
Data were analyzed using statistical methods (SPSS), with significance set at P < 0.05. Techniques included UV-vis spectroscopy, fluorescence imaging, flow cytometry, and histological analysis.
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NMR Spectrometer
500 MHz
Bruker
Measure 1H NMR spectra for structural analysis of polymers.
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XRD Diffractometer
D/MAX2500V/PC
Rigaku
Acquire X-ray diffraction patterns for crystalline structure analysis.
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Rheometer
Kinexus lab+
Malvern
Investigate rheological behavior of hydrogels.
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SEM
S-4800
Hitachi
Image microstructure of hydrogel matrices.
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CLSM
LSM 710
Carl Zeiss
Perform confocal laser scanning microscopy for morphological and cellular studies.
ZEISS LSM 990 Spectral Multiplex
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FL Microscope
IX51
Olympus
Visualize fluorescently stained cells for viability and ROS detection.
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Multimodal Imaging System
FX PRO
Bruker
Perform in vivo NIR fluorescence imaging for drug retention studies.
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FT-IR Spectrometer
Nexus 470
Nicolet
Record Fourier transform infrared spectra for material characterization.
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TGA System
Q500
TGA
Perform thermogravimetric analyses to determine composition.
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UV-vis-NIR Spectrophotometer
Perkin-Elmer
Obtain UV-vis-NIR absorption spectra for photothermal property assessment.
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Infrared Thermal Camera
MAG30
Shanghai Magnity Electronics Technology Co., Ltd.
Record real-time thermal images during photothermal experiments.
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NIR Laser
808 nm
Beijing Sitong Boyuan Co., Ltd.
Irradiate samples for photothermal and photodynamic therapy.
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Flow Cytometer
FACSverse
BD
Analyze cellular uptake and other cellular parameters.
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Microplate Reader
Infinite M1000
TECAN
Measure absorbance for MTT assays and other spectrophotometric analyses.
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Optical Inverted Microscope
MF53
Guangzhou Mingmei Industrial Co., Ltd
Acquire histological images for tissue analysis.
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Milli-Q Apparatus
Millipore
Purify water for experimental use.
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