Journal of Materials Science

, Volume 53, Issue 9, pp 6665–6680 | Cite as

Cytotoxicity, genotoxicity and uptake detection of folic acid-functionalized green upconversion nanoparticles Y2O3/Er3+, Yb3+ as biolabels for cancer cells

Electronic materials


Upconversion nanoparticles (UCNPs) have been used as biolabels for cancer cells due to their ability to absorb near-infrared photons and upconvert them into visible radiation. We reported the synthesis of UCNPs Y2O3/Yb3+, Er3+ (1, 1 mol%), which upon excitation with infrared photons (λ = 980 nm) emit green color with a maximum peak centered at λ = 550 nm. UCNPs were functionalized with folic acid (UCNPs-NH2-FA) and analyzed by transmission electron microscopy, Fourier transform infrared spectroscopy, XRD, DLS and photoluminescence measurements. UCNPs-NH2-FA had a particle size of 70 ± 10 nm and exhibit a good luminescence spectrum in comparison with bare UCNPs. Cytotoxicity of different concentrations of bare and functionalized UCNPs was measured with the MTT assay in three cancer cell lines: human cervical adenocarcinoma (HeLa) and human breast adenocarcinoma cells (MDA-MB-231 and MCF-7). Some concentrations of bare UCNPs were cytotoxic for cells; however, after been functionalized, UCNPs resulted to be non-cytotoxic. Genotoxicity of bare and functionalized UCNPs was performed by the comet assay, and no DNA damage was found for any concentration. The internalization of UCNPs-NH2-FA into cancer cells was confirmed by confocal microscopy showing a cytoplasmic fluorescence signal. UCNPs-NH2-FA were used to detect cancer cells in suspension by flow cytometry, with a specific green fluorescent signal for effective detection of cells. These results confirm that functionalized UCNPs can be used without any cytotoxic or genotoxic effects for bioimaging to detect and visualize cancer cells.



The authors wish to acknowledge financial support from DGAPA-UNAM Grant No. 109913 and CONACYT Project No. 269071. DGAPA-UNAM Grant No. 111017 and CONACYT Project Nos. 269071 and 232608. The authors acknowledge the technical support provided by E. Aparicio, F. Ruiz. M. Ponce, Dr. Katrin Quester, Dr. F. Castillón and Dr. Ruben D. Cadena Nava. The authors are grateful with the facilities provided by Dr. Rosa Mouriño at the Centro de Microscopía Avanzada (CEMIAD) of CICESE in the use of confocal microscopy and Dr. Olga Callejas for her technical assistance in imaging capture. Karla Juarez-Moreno is a member of the International Network of Bionanotechnology with impact in Biomedicine, Food and Biosafety (Funded by CONACYT Project 279889).

Compliance with ethical standards

Conflicts of interest

The authors declare that they have no conflicts of interest.

Supplementary material

10853_2017_1946_MOESM1_ESM.docx (96 kb)
Supplementary material 1 (DOCX 96 kb)


  1. 1.
    Lin M, Zhao Y, Wang S et al (2012) Recent advances in synthesis and surface modification of lanthanide-doped upconversion nanoparticles for biomedical applications. Biotechnol Adv 30:1551–1561. CrossRefGoogle Scholar
  2. 2.
    Wang M, Abbineni G, Clevenger A et al (2011) Upconversion nanoparticles: synthesis, surface modification and biological applications. Nanomed Nanotechnol Biol Med 7:710–729. CrossRefGoogle Scholar
  3. 3.
    Matsuura D (2002) Red, green, and blue upconversion luminescence of trivalent-rare-earth ion-doped Y2O3 nanocrystals. Appl Phys Lett 81:4526. CrossRefGoogle Scholar
  4. 4.
    Yang Y (2014) Upconversion nanophosphors for use in bioimaging, therapy, drug delivery and bioassays. Microchim, ActaGoogle Scholar
  5. 5.
    Shen J, Sun L-D, Yan C-H (2008) Luminescent rare earth nanomaterials for bioprobe applications. Dalt Trans. Google Scholar
  6. 6.
    Blasse G, Grabmaier B (1994) Luminescent materials, 1st edn. Springer, BerlinCrossRefGoogle Scholar
  7. 7.
    Chen GY, Liu Y, Zhang ZG et al (2007) Four-photon upconversion induced by infrared diode laser excitation in rare-earth-ion-doped Y2O3 nanocrystals. Chem Phys Lett 448:127–131. CrossRefGoogle Scholar
  8. 8.
    Kong W, Shan J, Ju Y (2010) Flame synthesis and effects of host materials on Yb3+/Er3+ co-doped upconversion nanophosphors. Mater Lett. Google Scholar
  9. 9.
    Sounderya N, Zhang Y (2009) Upconversion nanoparticles for imaging cells. In: IFMBE proceedings, pp 1741–1744Google Scholar
  10. 10.
    Anderson RR, Parrish JA (1981) The optics of human skin. J Invest Dermatol 77:13–19. CrossRefGoogle Scholar
  11. 11.
    Wang C, Tao H, Cheng L, Liu Z (2011) Near-infrared light induced in vivo photodynamic therapy of cancer based on upconversion nanoparticles. Biomaterials 32:6145–6154. CrossRefGoogle Scholar
  12. 12.
    Alexis F, Rhee J-WW, Richie JP et al (2008) New frontiers in nanotechnology for cancer treatment. Urol Oncol Semin Orig Investig 26:74–85. CrossRefGoogle Scholar
  13. 13.
    Sudimack J, Lee RJ (2000) Targeted drug delivery via the folate receptor. Adv Drug Deliv Rev 41:147–162. CrossRefGoogle Scholar
  14. 14.
    Lu Y, Sega E, Leamon CP, Low PS (2004) Folate receptor-targeted immunotherapy of cancer: mechanism and therapeutic potential. Adv Drug Deliv Rev 56:1161–1176. CrossRefGoogle Scholar
  15. 15.
    Yee K, Seow E, Zhang Y, Chyn Y (2013) Biomaterials targeting CCL21 e folic acid e upconversion nanoparticles conjugates to folate receptor-a expressing tumor cells in an endothelial-tumor cell bilayer model. Biomaterials 34:4860–4871. CrossRefGoogle Scholar
  16. 16.
    Rijnboutt S, Jansen G, Posthuma G et al (1996) Endocytosis of GPI-linked membrane folate receptor-alpha. J Cell Biol 132:35–47. CrossRefGoogle Scholar
  17. 17.
    Wu M, Gunning W, Ratnam M (1999) Expression of folate receptor type alpha in relation to cell type, malignancy, and differentiation in ovary, uterus, and cervix. Cancer Epidemiol Biomark Prev 8:775–782Google Scholar
  18. 18.
    Taxak VB, Khatkar SP, Han S-D et al (2009) Tartaric acid-assisted sol–gel synthesis of Y2O3:Eu3+ nanoparticles. J Alloys Compd 469:224–228. CrossRefGoogle Scholar
  19. 19.
    Chávez DH, Contreras OE, Hirata GA (2016) Synthesis and upconversion luminescence of nanoparticles Y2O3 and Gd2O3 Co-doped with Yb3+ and Er3+. Nanomater Nanotechnol. Google Scholar
  20. 20.
    Stöber W, Fink A, Bohn E (1968) Controlled growth of monodisperse silica spheres in the micron size range. J Colloid Interface Sci 26:62–69. CrossRefGoogle Scholar
  21. 21.
    Chavez DH, Juarez-Moreno K, Hirata GA (2016) Aminosilane functionalization and cytotoxicity effects of upconversion nanoparticles Y2O3 and Gd2O3 co-doped with Yb3+ and Er3+. Nanobiomedicine. Google Scholar
  22. 22.
    De Mello JC, Wittmann HF, Friend RH (1997) An improved experimental determination of external photoluminescence quantum efficiency. Adv Mater 9:230–232. CrossRefGoogle Scholar
  23. 23.
    Mosmann T (1983) Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 65:55–63CrossRefGoogle Scholar
  24. 24.
    Singh NP, McCoy MT, Tice RR, Schneider EL (1988) A simple technique for quantitation of low levels of DNA damage in individual cells. Exp Cell Res 175:184–191CrossRefGoogle Scholar
  25. 25.
    Speit G, Hartmann A (2006) The comet assay: a sensitive genotoxicity test for the detection of DNA damage and repair. Methods Mol Biol 314:275–286. CrossRefGoogle Scholar
  26. 26.
    Sánchez-Sánchez L, Tapia-Moreno A, Juarez-Moreno K et al (2015) Design of a VLP-nanovehicle for CYP450 enzymatic activity delivery. J Nanobiotechnol 13:1–10. CrossRefGoogle Scholar
  27. 27.
    Mader HS, Kele P, Saleh SM, Wolfbeis OS (2010) Upconverting luminescent nanoparticles for use in bioconjugation and bioimaging. Curr Opin Chem Biol 14:582–596. CrossRefGoogle Scholar
  28. 28.
    Wang M, Mi C, Zhang Y et al (2009) NIR-responsive silica-coated NaYbF 4:Er/Tm/Ho upconversion fluorescent nanoparticles with tunable emission colors and their applications in immunolabeling and fluorescent imaging of cancer cells. J Phys Chem C 113:19021–19027. CrossRefGoogle Scholar
  29. 29.
    Chatterjee DK, Rufaihah AJ, Zhang Y (2008) Upconversion fluorescence imaging of cells and small animals using lanthanide doped nanocrystals. Biomaterials 29:937–943. CrossRefGoogle Scholar
  30. 30.
    Chen G, Qiu H, Prasad PN, Chen X (2014) Upconversion nanoparticles: design, nanochemistry, and applications in theranostics. Chem Rev 114:5161–5214. CrossRefGoogle Scholar
  31. 31.
    Low PS, Henne WA, Doorneweerd DD (2008) Discovery and development of folic-acid-based receptor targeting for imaging and therapy of cancer and inflammatory diseases. Acc Chem Res 41:120–129. CrossRefGoogle Scholar
  32. 32.
    Ai J, Xu Y, Li D et al (2012) Folic acid as delivery vehicles: targeting folate conjugated fluorescent nanoparticles to tumors imaging. Talanta 101:32–37. CrossRefGoogle Scholar
  33. 33.
    Davis ME, Chen ZG, Shin DM (2008) Nanoparticle therapeutics: an emerging treatment modality for cancer. Nat Rev Drug Discov 7:771–782. CrossRefGoogle Scholar
  34. 34.
    Yoon SN, Ku J-L, Shin Y-K et al (2007) Hereditary nonpolyposis colorectal cancer in endometrial cancer patients. Int J Cancer 122:1077–1081. CrossRefGoogle Scholar
  35. 35.
    Karlsson HL, Di Bucchianico S, Collins AR, Dusinska M (2015) Can the comet assay be used reliably to detect nanoparticle-induced genotoxicity? Environ Mol Mutagen 56:82–96. CrossRefGoogle Scholar
  36. 36.
    Cheng L, Yang K, Shao M et al (2011) In vivo pharmacokinetics, long-term biodistribution and toxicology study of functionalized upconversion nanoparticles in mice. Nanomedicine 6(8):1327–1340. CrossRefGoogle Scholar
  37. 37.
    Shen J, Zhao L, Han G (2013) Lanthanide-doped upconverting luminescent nanoparticle platforms for optical imaging-guided drug delivery and therapy. Adv Drug Deliv Rev 65:744–755. CrossRefGoogle Scholar
  38. 38.
    Hemmer E, Yamano T, Kishimoto H et al (2013) Cytotoxic aspects of gadolinium oxide nanostructures for up-conversion and NIR bioimaging. Acta Biomater 9:4734–4743. CrossRefGoogle Scholar
  39. 39.
    Cheng L, Yang K, Li Y et al (2011) Facile preparation of multifunctional upconversion nanoprobes for multimodal imaging and dual-targeted photothermal therapy. Angew Chemie 123:7523–7528. CrossRefGoogle Scholar
  40. 40.
    Subik K, Lee J-F, Baxter L et al (2010) The expression patterns of ER, PR, HER2, CK5/6, EGFR, Ki-67 and AR by immunohistochemical analysis in breast cancer cell lines. Breast Cancer (Auckl) 4:35–41Google Scholar
  41. 41.
    Qian J, Wang D, Cai F et al (2012) Photosensitizer encapsulated organically modified silica nanoparticles for direct two-photon photodynamic therapy and in vivo functional imaging. Biomaterials 33:4851–4860. CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Centro de Enseñanza Técnica y SuperiorEnsenadaMexico
  2. 2.Center of Nanosciences and NanotechnologyAutonomus National University of MexicoEnsenadaMexico
  3. 3.Centro de Nanociencias y NanotecnologíaUniversidad Nacional Autónoma de MéxicoEnsenadaMexico
  4. 4.Facultad de Ciencias QuímicasUniversidad de ConcepciónConcepciónChile
  5. 5.Departamento de FisicoquímicaCentro de Nanociencias y Nanotecnología UNAMEnsenadaMexico

Personalised recommendations