Cytotoxicity, genotoxicity and uptake detection of folic acid-functionalized green upconversion nanoparticles Y2O3/Er3+, Yb3+ as biolabels for cancer cells
- 12 Downloads
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.
- 4.Yang Y (2014) Upconversion nanophosphors for use in bioimaging, therapy, drug delivery and bioassays. Microchim, ActaGoogle Scholar
- 9.Sounderya N, Zhang Y (2009) Upconversion nanoparticles for imaging cells. In: IFMBE proceedings, pp 1741–1744Google Scholar
- 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. https://doi.org/10.1016/j.biomaterials.2011.05.007 CrossRefGoogle Scholar
- 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. https://doi.org/10.1016/j.biomaterials.2013.03.029 CrossRefGoogle Scholar
- 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
- 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. https://doi.org/10.1021/jp906394z CrossRefGoogle Scholar
- 29.Chatterjee DK, Rufaihah AJ, Zhang Y (2008) Upconversion fluorescence imaging of cells and small animals using lanthanide doped nanocrystals. Biomaterials 29:937–943. https://doi.org/10.1016/j.biomaterials.2007.10.051 CrossRefGoogle Scholar
- 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.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. https://doi.org/10.1016/j.biomaterials.2012.02.053 CrossRefGoogle Scholar