RGDS- and TAT-Conjugated Upconversion of NaYF4:Yb3+/Er3+&SiO2 Nanoparticles: In Vitro Human Epithelioid Cervix Carcinoma Cellular Uptake, Imaging, and Targeting

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• 作者：Uliana Kostiv ; Ilya Kotelnikov ; Vladimír Proks ; Miroslav Šlouf ; Jan Kučka ; Hana Engstová ; Petr Ježek ; Daniel Horák
• 刊名：ACS Applied Materials & Interfaces
• 出版年：2016
• 出版时间：August 10, 2016
• 年：2016
• 卷：8
• 期：31
• 页码：20422-20431
• 全文大小：624K
• 年卷期：0
• ISSN：1944-8252

文摘

Starting NaYF₄:Yb³⁺/Er³⁺ nanoparticles with size tuned from 24 to 33 nm were prepared by high-temperature coprecipitation of lanthanide chlorides in high-boiling organic solvents. To enhance colloidal stability in aqueous medium, an aminosilica shell was introduced on the surface by hydrolysis and condensation of tetramethyl orthosilicate and (3-aminopropyl)trimethoxysilane using a reverse microemulsion technique; to form alkyne groups, reaction with 4-pentynoic acid followed. Finally, the cell adhesive and cell penetrating azidopentanoyl-GGGRGDSGGGY-NH₂ (RGDS) and azidopentanoyl-GGGRKKRRQRRR-NH₂ (TAT) peptides were conjugated to the upconversion particles via Cu(I)-catalyzed alkyne–azide cycloaddition. The concentrations of the peptides bound to the nanoparticle surfaces and amount of adsorbed residual Cu(I) catalyst were determined using an ¹²⁵I-radiolabeled RGDS peptide and a ⁶⁴Cu(I)-doped catalyst, respectively. Targeting and uptake of the RGDS- and TAT-conjugated NaYF₄:Yb³⁺/Er³⁺&SiO₂ nanoparticles by human cervix carcinoma HeLa cells were monitored by confocal microscopy. RGDS-conjugated nanoparticle probes were mainly localized on the cell plasma membrane due to specific binding of the peptide to the corresponding integrins. In contrast, the TAT-conjugated nanoparticles were able to cross the cell membrane and accumulate in the cell cytoplasm. Thus, this new peptide bioconjugation approach supported both extra- and intracellular nanoparticle uptake, enabling targeting and imaging of the specific tumor phenotypes.