Journal
COLLOIDS AND SURFACES B-BIOINTERFACES
Volume 145, Issue -, Pages 870-877Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.colsurfb.2016.06.006
Keywords
Zinc oxide nanoparticles; Particle size; Physico-chemical property of ZnO; Cy5.5 conjugation; Fluorescent stability; Cellular uptake
Funding
- Nuclear R&D Program through the National Research Foundation of Korea - Ministry of Science, ICT and Future Planning [2015M2B2A4031430]
- Seoul Women's University
- Ministry of Public Administration & Security (MOIS), Republic of Korea [2016-화학-02] Funding Source: Korea Institute of Science & Technology Information (KISTI), National Science & Technology Information Service (NTIS)
- National Research Foundation of Korea [2015M2B2A4031430] Funding Source: Korea Institute of Science & Technology Information (KISTI), National Science & Technology Information Service (NTIS)
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We evaluated size dependent cellular uptake of ZnO nanoparticles utilizing stably introduced Cy5.5, which emits long-wavelength fluorescence. Through (3-aminopropyl)triethoxysilane modification, ZnO nanoparticles of different sizes (20 and 70 nm) were functionalized with amine moiety, which was further reacted with Cy5.5-N-hydroxylsuccinimide ester to make covalently conjugated Cy5.5 dye on ZnO nanoparticles. Field emission-scanning electron microscopic images revealed that average particle size as well as particle morphology of ZnO nanoparticles were not altered by Cy5.5 conjugation. Zeta potential measurement confirmed that the positive surface charge of ZnO nanoparticles was well preserved after successive conjugation reactions. Based on infrared, ultraviolet-visible light and photoluminescence spectroscopies, we verify that the Cy5.5 was stably introduced to ZnO nanoparticles without serious aggregation. Surface conjugated Cy5.5 showed high stability in deionized water, phosphate buffered saline and cell culture medium, showing less than 2% of release during 85 h. Confocal microscopy and fluorescence-activated cell sorting analysis demonstrated that smaller ZnO nanoparticles were more taken up in greater quantities by HaCaT cells. Moreover, systematic study on cellular uptake pathway showed that smaller ZnO nanoparticles were internalized into cells mainly by clathrin-mediated endocytosis, while larger ZnO nanoparticles entered cells via several pathways. (C) 2016 Elsevier B.V. All rights reserved.
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