研究目的
To develop a pH-sensitive and fluorescent drug delivery system using PEG-TPP to encapsulate paclitaxel, overcoming its poor water solubility and enhancing its antitumor activity through targeted delivery to tumors and mitochondria.
研究成果
The PEG-TPP-based micelles effectively encapsulated paclitaxel, demonstrated pH-sensitive release, targeted mitochondria in cancer cells, and showed enhanced antitumor activity both in vitro and in vivo compared to free PTX. The system is biodegradable, selective, and holds promise for improving nanomedicine applications in cancer therapy.
研究不足
The study notes that the poor penetrability of green fluorescence limited in vivo imaging of micelle distribution in living mice. Additionally, the stability of micelles was only tested for one week, and further optimization may be needed for longer-term applications.
1：Experimental Design and Method Selection:
The study involved synthesizing PEG-TPP as a carrier for paclitaxel micelles, using emulsion solvent evaporation method for preparation. Theoretical models include EPR effect for tumor targeting and pH sensitivity for controlled release.
2：Sample Selection and Data Sources:
Samples included PTX-loaded micelles, blank micelles, and free PTX. Cell lines used were K562, MCF-7, and Hela for in vitro studies, and MCF-7 and U87 tumor-bearing mice for in vivo studies.
3：List of Experimental Equipment and Materials:
Equipment included particle size instrument (Brookhaven, USA), TEM (HITACHI HT7700, Japan), HPLC (SHIMADZU LC-20A, Japan), LCFM (Olympus FV1000, Japan), microplate reader (Thermo, USA), living imaging system (PerkinElmer IVIS Spectrum, USA), and bright field microscopy (Olympus BX41). Materials included PEG(2000)-OH, Br-TPP, K2CO3, acetonitrile, dichloromethane, methanol, acetone, PTX (Saen chemical technology Co., Ltd.), pyrene, tween 80 (Aladdin), Mito Tracker Red, MTT dye, DMSO, and various buffers.
4：1). Materials included PEG(2000)-OH, Br-TPP, K2CO3, acetonitrile, dichloromethane, methanol, acetone, PTX (Saen chemical technology Co., Ltd.), pyrene, tween 80 (Aladdin), Mito Tracker Red, MTT dye, DMSO, and various buffers. Experimental Procedures and Operational Workflow:
4. Experimental Procedures and Operational Workflow: Synthesis of PEG-TPP via reaction in acetonitrile, purification by column chromatography. Preparation of micelles by dissolving PTX and PEG-TPP in acetone and adding to water. Characterization of micelles for size, morphology, CMC, stability, encapsulation efficiency, and in vitro release at different pH values. Cellular uptake and mitochondria targeting observed using LCFM. Cytotoxicity assessed via MTT assay. In vivo distribution and antitumor activity studied in tumor-bearing mice, with fluorescence imaging and histological analysis.
5：Data Analysis Methods:
Data analyzed using GraphPad Prism 5.0 for statistical analysis and chart generation. Particle size and polydispersity measured in triplicate. Cytotoxicity tests performed in triplicate.
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transmission electron microscopy
HT7700
HITACHI
Morphological evaluation of micelles
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HPLC system
LC-20A
SHIMADZU
Detecting the content of PTX in micelles
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laser confocal fluorescence microscopy
FV1000
Olympus
Observing cellular uptake and mitochondria targeting
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living imaging system
IVIS Spectrum
PerkinElmer
Detecting fluorescence distribution in excised organs
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bright field microscopy
BX41
Olympus
Observing sliced organs
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particle size instrument
Brookhaven
Measuring the hydrodynamic diameter of micelles
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microplate reader
Thermo
Reading optical density in MTT assay
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