期刊
NATURE PROTOCOLS
卷 11, 期 4, 页码 688-713出版社
NATURE PORTFOLIO
DOI: 10.1038/nprot.2016.035
关键词
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资金
- Agency for Science, Technology and Research (A*STAR) Biomedical Research Council [R-397-000-062-305, R-397-000-119-305]
- Biomedical Engineering Programme [R-397-000-128-305]
- National Medical Research Council (NMRC) [CBRG13nov052, R-397-000-199-511]
- National Natural Science Foundation of China [31328009]
- National University of Singapore
- NGS scholarship from NUS graduate school (NGS) for Integrative Sciences and Engineering, National University of Singapore
Photoactivation is a process in which light is used to 'activate' photolabile therapeutics. As a therapeutic strategy, its advantages are that it is noninvasive and that a high degree of spatial and temporal control is possible. However, conventional photoactivation techniques are hampered by the limited penetration depth of the UV and visible lights to which the photosensitive compounds are responsive. Here we describe a protocol for the use of upconversion nanoparticles (UCNs) as light transducers to convert deeply penetrating near-infrared (NIR) light to UV-visible wavelengths matching that of the absorption spectrum of photosensitive therapeutics. This allows the use of deep-penetrating and biologically friendly NIR light instead of low-penetrating and/or toxic visible or UV lights for photoactivation. In this protocol, we focus on two photoactivation applications: photodynamic therapy (PDT) and photoactivated control of gene expression. We describe how to prepare and characterize the UCNs, as well as how to check their function in biochemical assays and in cells. For both applications, the UCNs are coated with mesoporous silica for easy loading of the therapeutics. For PDT, the UCNs are coated with polyethylene glycol (PEG) for stabilization and folic acid for tumor targeting and then loaded with photosensitizers that would be expected to kill cells by singlet oxygen production; the nanoparticles are injected intravenously. For photoactivated control of gene expression, knockdown of essential tumor genes is achieved using UCNs loaded with caged nucleic acids, which are injected intratumorally. The whole process from nanoparticle synthesis to animal studies takes similar to 36 d.
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