Systematic evaluation of oligodeoxynucleotide binding and hybridization to modified multi-walled carbon nanotubes

Journal of Nanobiotechnology

Research

Bernd Schwenzer¹  Anika Kaufmann²  Kati Erdmann³  Susanne Füssel³  Christiane Rieger³  Darja Schendel³  Silke Hampel⁴  David Kunhardt⁴ 

[1] Chair of Biochemistry, Department of Chemistry, Technische Universität Dresden, Bergstraße 66, 01069, Dresden, Germany;Chair of Biochemistry, Department of Chemistry, Technische Universität Dresden, Bergstraße 66, 01069, Dresden, Germany;Department of Nanostructured Materials, Leibniz Institute for Polymer Research, Hohe Straße 6, 01069, Dresden, Germany;Department of Urology, University Hospital Carl Gustav Carus, Technische Universität Dresden, Fetscherstraße 74, 01307, Dresden, Germany;Leibniz Institute for Solid State and Materials Research Dresden, Helmholtzstraße 20, 01069, Dresden, Germany;

关键词: Antisense oligodeoxynucleotides;    Biocompatibility;    Bladder cancer;    Carbon nanotubes;    Carrier strand;    Functionalization;    Hybridization;    Mucoadhesion;    Urothelium;   

DOI  :  10.1186/s12951-017-0288-z

 received in 2017-03-07, accepted in 2017-07-08,  发布年份 2017

来源: Springer

【 摘 要 】

BackgroundIn addition to conventional chemotherapeutics, nucleic acid-based therapeutics like antisense oligodeoxynucleotides (AS-ODN) represent a novel approach for the treatment of bladder cancer (BCa). An efficient delivery of AS-ODN to the urothelium and then into cancer cells might be achieved by the local application of multi-walled carbon nanotubes (MWCNT). In the present study, pristine MWCNT and MWCNT functionalized with hydrophilic moieties were synthesized and then investigated regarding their physicochemical characteristics, dispersibility, biocompatibility, cellular uptake and mucoadhesive properties. Finally, their binding capacity for AS-ODN via hybridization to carrier strand oligodeoxynucleotides (CS-ODN), which were either non-covalently adsorbed or covalently bound to the different MWCNT types, was evaluated.ResultsPristine MWCNT were successfully functionalized with hydrophilic moieties (MWCNT-OH, -COOH, -NH2, -SH), which led to an improved dispersibility and an enhanced dispersion stability. A viability assay revealed that MWCNT-OH, MWCNT-NH2 and MWCNT-SH were most biocompatible. All MWCNT were internalized by BCa cells, whereupon the highest uptake was observed for MWCNT-OH with 40% of the cells showing an engulfment. Furthermore, all types of MWCNT could adhere to the urothelium of explanted mouse bladders, but the amount of the covered urothelial area was with 2–7% rather low. As indicated by fluorescence measurements, it was possible to attach CS-ODN by adsorption and covalent binding to functionalized MWCNT. Adsorption of CS-ODN to pristine MWCNT, MWCNT-COOH and MWCNT-NH2 as well as covalent coupling to MWCNT-NH2 and MWCNT-SH resulted in the best binding capacity and stability. Subsequently, therapeutic AS-ODN could be hybridized to and reversibly released from the CS-ODN coupled via both strategies to the functionalized MWCNT. The release of AS-ODN at experimental conditions (80 °C, buffer) was most effective from CS-ODN adsorbed to MWCNT-OH and MWCNT-NH2 as well as from CS-ODN covalently attached to MWCNT-COOH, MWCNT-NH2 and MWCNT-SH. Furthermore, we could exemplarily demonstrate that AS-ODN could be released following hybridization to CS-ODN adsorbed to MWCNT-OH at physiological settings (37 °C, urine).ConclusionsIn conclusion, functionalized MWCNT might be used as nanotransporters in antisense therapy for the local treatment of BCa.

【 授权许可】

CC BY   
© The Author(s) 2017