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was observed in an in vitro spheroid model system mimicking tumor tissues in vivo (Figure 9C). The Z-stack images of spheroids were obtained from slices with a 20-μm step size and showed red fluorescence. Based on the spheroid result, UCNPs@SiO2-NH2@FA/PEG/TCPP was predicted to penetrate into tumors in vivo. Despite the absence of in vivo tumor ablation, we believe that these oPhvaermralcleuitnicsv20it2r0o, 12d,a1t1a02demonstrate the great potential for tumor targeting and penetration o1f3 ofu1r7 UCNPs platform. Figure 9. (A) Two-dimensional monolayer images of HeLa cells treated with core-shell upconverting Figure 9. (A) Two-dimensional monolayer images of HeLa cells treated with core-shell nanoparticles surface-modified with silica, PEG-folic acid (FA/PEG), and TCPP upconverting nanoparticles surface-modified with silica, PEG-folic acid (FA/PEG), and TCPP (UCNP@SiO2@FA/PEG/TCPP) at predetermined time points (blue: nuclei, green: lysosomes, red: (UCNP@SiO2 @FA/PEG/TCPP) at predetermined time points (blue: nuclei, green: lysosomes, red: TCPP). (B) Bio-TEM images of HeLa cells treated with UCNP@SiO2@FA/PEG/TCPP. (C) In vitro spheroid model images (Z-stack of ten 20-μm slices) for predicting the cellular uptake of UCNP conjugates into three-dimensional tumor tissues. 4. Conclusions In summary, highly singlet oxygen-generating upconverting nanoparticles respectively doped with erbium and lutetium for the core and shell (UCNPs@SiO2-NH2@FA/PEG/TCPP) were fabricated. These nanoparticles were ~50 nm in size and strongly emitted red fluorescence with little green fluorescence, which was considered optimal for the activation of a 660 nm-responsive-photodynamic TCPP agent. The UCNPs generated singlet oxygen and thus killed HeLa cells under 808 nm laser irradiation and normoxic conditions. Furthermore, the UCNPs were well-internalized into the HeLa cell monolayer and three-dimensional cell spheroids, presumably due to the folic acid ligand and small particle size. These overall results suggest that UCNPs could be a potential therapeutic platform for ablating tumors. Author Contributions: Conceptualization: Y.S.Y., K.L. and X.T.L.; Methodology, K.L., H.K.K. and N.T.N.; Software: H.K.K.; Validation: X.T.L. and H.K.K.; Formal analysis: K.L. and X.T.L.; Investigation: E.S.L., K.T.O.; Resources: H.-G.C.; Data curation: H.K.K., X.T.L. and E.S.L., Y.S.Y.; Writing-original draft preparation: K.L. and Y.S.Y.; Writing-review and editing: E.S.L., K.T.O., Y.S.Y. and H.-G.C.; Visualization: Y.S.Y.; Supervision: Y.S.Y.; Project administration: Y.S.Y.; Funding acquisition: Y.S.Y. All authors have read and agreed to the published version of the manuscript. Funding: This work was supported by National Research Foundation of Korea (NRF) grants funded by the Korean government (MSIT) (No. NRF-2019R1A2C2085292 and No. NRF-2019R1A5A2027340). Conflicts of Interest: The authors declare no conflict of interest. References 1. Castano, A.P.; Mroz, P.; Hamblin, M.R. Photodynamic therapy and anti-tumour immunity. Nat. Rev. Cancer 2006, 6, 535–545. [CrossRef] [PubMed] 2. Phuong, P.T.T.; Lee, S.; Lee, C.; Seo, B.; Park, S.; Oh, K.T.; Lee, E.S.; Choi, H.-G.; Shin, B.S.; Youn, Y.S. Beta-carotene-bound albumin nanoparticles modified with chlorin e6 for breast tumor ablation based on photodynamic therapy. Colloids Surf. B Biointerfaces 2018, 171, 123–133. [CrossRef] [PubMed]PDF Image | Red LED Erbium Upconverting Nanoparticles v Cervical Cancer
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