Abstract
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 ∼36 d.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$259.00 per year
only $21.58 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Jochum, F.D. & Theato, P. Temperature- and light-responsive smart polymer materials. Chem. Soc. Rev. 42, 7468–7483 (2013).
Agasti, S.S. et al. Photoregulated release of caged anticancer drugs from gold nanoparticles. J. Am. Chem. Soc. 131, 5728–5729 (2009).
Tong, X., Wang, G., Soldera, A. & Zhao, Y. How can azobenzene block copolymer vesicles be dissociated and reformed by light? J. Phys. Chem. B 109, 20281–20287 (2005).
Katz, J.S. & Burdick, J.A. Light-responsive biomaterials: development and applications. Macromol. Biosci. 10, 339–348 (2010).
Wang, F., Banerjee, D., Liu, Y., Chen, X. & Liu, X. Upconversion nanoparticles in biological labeling, imaging, and therapy. Analyst 135, 1839–1854 (2010).
Pawlicki, M., Collins, H.A., Denning, R.G. & Anderson, H.L. Two-photon absorption and the design of two-photon dyes. Angew. Chem. Int. Ed. 48, 3244–3266 (2009).
Dong, B. et al. Multifunctional NaYF4:Yb3+,Er3+@Ag core/shell nanocomposites: integration of upconversion imaging and photothermal therapy. J. Mater. Chem. 21, 6193 (2011).
Zhang, P., Steelant, W., Kumar, M. & Scholfield, M. Versatile photosensitizers for photodynamic therapy at infrared excitation. J. Am. Chem. Soc. 129, 4526–4527 (2007).
Chatterjee, D.K. & Yong, Z. Upconverting nanoparticles as nanotransducers for photodynamic therapy in cancer cells. Nanomedicine 3, 73–82 (2008).
Ungun, B. et al. Nanofabricated upconversion nanoparticles for photodynamic therapy. Opt. Express 17, 80–86 (2009).
Qian, H., Guo, H., Ho, P., Mahendran, R. & Zhang, Y. Mesoporous-silica-coated up-conversion fluorescent nanoparticles for photodynamic therapy. Small 5, 2285–2290 (2009).
Guo, H.C., Qian, H.S., Idris, N.M. & Zhang, Y. Singlet oxygen-induced apoptosis of cancer cells using upconversion fluorescent nanoparticles as a carrier of photosensitizer. Nanomedicine 6, 486–495 (2010).
Shan, J.N. et al. Pegylated composite nanoparticles containing upconverting phosphors and meso-tetraphenyl porphine (TPP) for photodynamic therapy. Adv. Funct. Mater. 21, 2488–2495 (2011).
Wang, C., Tao, H.Q., Cheng, L. & Liu, Z. Near-infrared light induced in vivo photodynamic therapy of cancer based on upconversion nanoparticles. Biomaterials 32, 6145–6154 (2011).
Cui, S.S. et al. Amphiphilic chitosan modified upconversion nanoparticles for in vivo photodynamic therapy induced by near-infrared light. J. Mater. Chem. 22, 4861–4873 (2012).
Lim, M.E., Lee, Y.L., Zhang, Y. & Chu, J.J.H. Photodynamic inactivation of viruses using upconversion nanoparticles. Biomaterials 33, 1912–1920 (2012).
Liu, K. et al. Covalently assembled NIR nanoplatform for simultaneous fluorescence imaging and photodynamic therapy of cancer cells. ACS Nano 6, 4054–4062 (2012).
Zhao, Z.X. et al. Multifunctional core-shell upconverting nanoparticles for imaging and photodynamic therapy of liver cancer cells. Chemistry 7, 830–837 (2012).
Chen, F. et al. A uniform sub-50 nm-sized magnetic/upconversion fluorescent bimodal imaging agent capable of generating singlet oxygen by using a 980-nm laser. Chemistry 18, 7082–7090 (2012).
Zhou, A.G., Wei, Y.C., Wu, B.Y., Chen, Q. & Xing, D. Pyropheophorbide A and c(RGDyK) comodified chitosan-wrapped upconversion nanoparticle for targeted near-infrared photodynamic therapy. Mol. Pharm. 9, 1580–1589 (2012).
Qiao, X.F. et al. Triple-functional core-shell structured upconversion luminescent nanoparticles covalently grafted with photosensitizer for luminescent, magnetic resonance imaging and photodynamic therapy in vitro. Nanoscale 4, 4611–4623 (2012).
Xu, Q.C. et al. Anti-cAngptl4 Ab-conjugated N-TiO2/NaYF4:Yb,Tm nanocomposite for near infrared-triggered drug release and enhanced targeted cancer cell ablation. Adv. Healthc. Mater. 1, 470–474 (2012).
Idris, N.M. et al. In vivo photodynamic therapy using upconversion nanoparticles as remote-controlled nanotransducers. Nat. Med. 18, 1580–1585 (2012).
Park, Y. et al. Theranostic probe based on lanthanide-doped nanoparticles for simultaneous in vivo dual-modal imaging and photodynamic therapy. Adv. Mater. 24, 5755–5761 (2012).
Xiao, Q.B. et al. Novel multifunctional NaYF4:Er3+,Yb3+/PEGDA hybrid microspheres: NIR-light-activated photopolymerization and drug delivery. Chem. Commun. 49, 1527–1529 (2013).
Cui, S.S. et al. In vivo targeted deep-tissue photodynamic therapy based on near-infrared light triggered upconversion nanoconstruct. ACS Nano 7, 676–688 (2013).
Liu, X. et al. Separately doped upconversion-C60 nanoplatform for NIR imaging-guided photodynamic therapy of cancer cells. Chem. Commun. (Camb.) 49, 3224–3226 (2013).
Zeng, L.Y. et al. Multifunctional photosensitizer-conjugated core-shell Fe3O4@NaYF4:Yb/Er nanocomplexes and their applications in T-2-weighted magnetic resonance/upconversion luminescence imaging and photodynamic therapy of cancer cells. RSC Adv. 3, 13915–13925 (2013).
Tian, G. et al. Red-emitting upconverting nanoparticles for photodynamic therapy in cancer cells under near-infrared excitation. Small 9, 1929–1938 (2013).
Wang, C. et al. Imaging-guided pH-sensitive photodynamic therapy using charge reversible upconversion nanoparticles under near-infrared light. Adv. Funct. Mater. 23, 3077–3086 (2013).
Wang, H.J., Shrestha, R. & Zhang, Y. Encapsulation of photosensitizers and upconversion nanocrystals in lipid micelles for photodynamic therapy. Part. Part. Syst. Char. 31, 228–235 (2014).
Jin, S. et al. A new near infrared photosensitizing nanoplatform containing blue-emitting up-conversion nanoparticles and hypocrellin A for photodynamic therapy of cancer cells. Nanoscale 5, 11910–11918 (2013).
Shimoyama, A. et al. Access to a novel near-infrared photodynamic therapy through the combined use of 5-aminolevulinic acid and lanthanide nanoparticles. Photodiagnosis Photodyn. Ther. 10, 607–614 (2013).
Yuan, Q. et al. Targeted bioimaging and photodynamic therapy nanoplatform using an aptamer-guided G-quadruplex DNA carrier and near-infrared light. Angew. Chem. Int. Ed. 52, 13965–13969 (2013).
Xia, L. et al. An upconversion nanoparticle—zinc phthalocyanine based nanophotosensitizer for photodynamic therapy. Biomaterials 35, 4146–4156 (2014).
Wang, X., Yang, C.X., Chen, J.T. & Yan, X.P. A dual-targeting upconversion nanoplatform for two-color fluorescence imaging-guided photodynamic therapy. Anal. Chem. 86, 3263–3267 (2014).
Chen, Q. et al. Protein modified upconversion nanoparticles for imaging-guided combined photothermal and photodynamic therapy. Biomaterials 35, 2915–2923 (2014).
Idris, N.M., Lucky, S.S., Li, Z.Q., Huang, K. & Zhang, Y. Photoactivation of core-shell titania-coated upconversion nanoparticles and their effect on cell death. J. Mater. Chem. B 2, 7017–7026 (2014).
Lucky, S.S. et al. Titania coated upconversion nanoparticles for near-infrared light triggered photodynamic therapy. ACS Nano 9, 191–205 (2015).
Zhang, L. et al. Inorganic photosensitizer coupled Gd-based upconversion luminescent nanocomposites for in vivo magnetic resonance imaging and near-infrared-responsive photodynamic therapy in cancers. Biomaterials 44, 82–90 (2015).
Fan, W.P. et al. A smart upconversion-based mesoporous silica nanotheranostic system for synergetic chemo-/radio-/photodynamic therapy and simultaneous MR/UCL imaging. Biomaterials 35, 8992–9002 (2014).
Scaffidi, J.P., Gregas, M.K., Lauly, B., Zhang, Y. & Vo-Dinh, T. Activity of Psoralen-functionalized nanoscintillators against cancer cells upon X-ray excitation. ACS Nano 5, 4679–4687 (2011).
Bulin, A.L. et al. X-ray-induced singlet oxygen activation with nanoscintillator-coupled porphyrins. J. Phys. Chem. C 117, 21583–21589 (2013).
Tang, Y.G., Hu, J., Elmenoufy, A.H. & Yang, X.L. Highly efficient FRET system capable of deep photodynamic therapy established on X-ray excited mesoporous LaF3:Tb scintillating nanoparticles. ACS Appl. Mater. Interfaces 7, 12261–12269 (2015).
Zou, X.J. et al. X-ray-induced nanoparticle-based photodynamic therapy of cancer. Nanomedicine 9, 2339–2351 (2014).
Chen, W. Nanoparticle self-lighting photodynamic therapy for cancer treatment. J. Biomed. Nanotechnol. 4, 369–376 (2008).
Chen, W. & Zhang, J. Using nanoparticles to enable simultaneous radiation and photodynamic therapies for cancer treatment. J. Nanosci. Nanotechnol. 6, 1159–1166 (2006).
Lu, T., Shao, P., Mathew, I., Sand, A. & Sun, W.F. Synthesis and photophysics of benzotexaphyrin: a near-infrared emitter and photosensitizer. J. Am. Chem. Soc. 130, 15782–15783 (2008).
Luan, L.Q. et al. A naphthalocyanine based near-infrared photosensitizer: synthesis and in vitro photodynamic activities. Bioorg. Med. Chem. Lett. 23, 3775–3779 (2013).
Yang, Y. et al. Thienopyrrole-expanded BODIPY as a potential NIR photosensitizer for photodynamic therapy. Chem. Commun. 49, 3940–3942 (2013).
Jayakumar, M.K.G. et al. Near-infrared-light-based nano-platform boosts endosomal escape and controls gene knockdown in vivo. ACS Nano 8, 4848–4858 (2014).
Jayakumar, M.K.G., Idris, N.M. & Zhang, Y. Remote activation of biomolecules in deep tissues using near-infrared-to-UV upconversion nanotransducers. Proc. Natl. Acad. Sci. USA 109, 8483–8488 (2012).
Li, Z.Q. & Zhang, Y. An efficient and user-friendly method for the synthesis of hexagonal-phase NaYF4 : Yb, Er/Tm nanocrystals with controllable shape and upconversion fluorescence. Nanotechnology 19, 345606–345610 (2008).
Li, Z.Q., Zhang, Y. & Jiang, S. Multicolor core/shell-structured upconversion fluorescent nanoparticles. Adv. Mater. 20, 4765–4769 (2008).
Li, C.X., Liu, J.L., Alonso, S., Li, F.Y. & Zhang, Y. Upconversion nanoparticles for sensitive and in-depth detection of Cu2+ ions. Nanoscale 4, 6065–6071 (2012).
Liu, J.A. et al. Controlled synthesis of uniform and monodisperse upconversion core/mesoporous silica shell nanocomposites for bimodal imaging. Chemistry 18, 2335–2341 (2012).
Jokerst, J.V., Miao, Z., Zavaleta, C., Cheng, Z. & Gambhir, S.S. Affibody-functionalized gold-silica nanoparticles for Raman molecular imaging of the epidermal growth factor receptor. Small 7, 625–633 (2011).
Hermanson, G.T. Bioconjugate Techniques 3rd ed. (Elsevier/Academic Press, 2013).
Pan, L.M. et al. Nuclear-targeted drug delivery of TAT peptide-conjugated monodisperse mesoporous silica nanoparticles. J. Am. Chem. Soc. 134, 5722–5725 (2012).
Yang, D.J. et al. Super hydrophobic mesoporous silica with anchored methyl groups on the surface by a one-step synthesis without surfactant template. J. Phys. Chem. C 111, 999–1004 (2007).
Lu, J., Liong, M., Zink, J.I. & Tamanoi, F. Mesoporous silica nanoparticles as a delivery system for hydrophobic anticancer drugs. Small 3, 1341–1346 (2007).
Gnanasammandhan, M.K., Bansal, A. & Zhang, Y. Light-activated endosomal escape using upconversion nanoparticles for enhanced delivery of drugs, Proc. SPIE 8594, Nanoscale Imaging, Sensing, and Actuation for Biomedical Applications X 85940B (February 19, 2013), doi:10.1117/12.2003720 (2013).
Acknowledgements
We thank S.S. Lucky for the helpful discussion. We also thank S. Jayakumar for technical support in the schematics. Y.Z. received funding support from the Agency for Science, Technology and Research (A*STAR) Biomedical Research Council (grant nos. R-397-000-062-305 and R-397-000-119-305), the Biomedical Engineering Programme (grant no. R-397-000-128-305), the National Medical Research Council (NMRC, grant nos. CBRG13nov052 and R-397-000-199-511), the National Natural Science Foundation of China (grant no. 31328009) and the National University of Singapore. A.B. is a recipient of the NGS scholarship from NUS graduate school (NGS) for Integrative Sciences and Engineering, National University of Singapore.
Author information
Authors and Affiliations
Contributions
All authors conceived and designed the experiments, analyzed the data and wrote the paper. K.H. prepared the UCN constructs and their characterizations. N.M.I. performed the photosensitizer loading into UCN constructs and validated them in in vitro and in vivo PDT studies. M.K.G. and A.B. performed the nucleic acid loading into UCN constructs and validated them in in vitro and in vivo studies on photoactivated control of gene expression. Y.Z. supervised the project. All authors commented on the manuscript at all stages.
Corresponding author
Ethics declarations
Competing interests
The authors declare no competing financial interests.
Rights and permissions
About this article
Cite this article
Gnanasammandhan, M., Idris, N., Bansal, A. et al. Near-IR photoactivation using mesoporous silica–coated NaYF4:Yb,Er/Tm upconversion nanoparticles. Nat Protoc 11, 688–713 (2016). https://doi.org/10.1038/nprot.2016.035
Published:
Issue Date:
DOI: https://doi.org/10.1038/nprot.2016.035
This article is cited by
-
Targeted theranostic photoactivation on atherosclerosis
Journal of Nanobiotechnology (2021)
-
Near-infrared manipulation of multiple neuronal populations via trichromatic upconversion
Nature Communications (2021)
-
Dye-doped silica nanoparticles: synthesis, surface chemistry and bioapplications
Cancer Nanotechnology (2020)
-
Recent progress in the development of upconversion nanomaterials in bioimaging and disease treatment
Journal of Nanobiotechnology (2020)
-
Near-infrared photoresponsive drug delivery nanosystems for cancer photo-chemotherapy
Journal of Nanobiotechnology (2020)
Comments
By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.