Two-Step Mechanism of Cellular Uptake of Cationic Gold Nanoparticles Modified by (16-Mercaptohexadecyl)trimethylammonium Bromide.
Cationic colloidal gold nanorods (GNRs) have a great potential as a theranostic tool for diverse medical applications. GNRs' properties such as cellular internalization and stability are determined by physicochemical characteristics of their surface coating. GNRs modified by (16-mercaptohexadecyl)trimethylammonium bromide (MTAB),MTABGNRs, show excellent cellular uptake. Despite their promise for biomedicine, however, relatively little is known about the cellular pathways that facilitate the uptake of GNRs, their subcellular fate and intracellular persistence. Here we studied the mechanism of cellular internalization and long-term fate of GNRs coated with MTAB, for which the synthesis was optimized to give higher yield, in various human cell types including normal diploid versus cancerous, and dividing versus nondividing (senescent) cells. The process ofMTABGNRs internalization into their final destination in lysosomes proceeds in two steps: (1) fast passive adhesion to cell membrane mediated by sulfated proteoglycans occurring within minutes and (2) slower active transmembrane and intracellular transport of individual nanorods via clathrin-mediated endocytosis and of aggregated nanorods via macropinocytosis. The expression of sulfated proteoglycans was the major factor determining the extent of uptake by the respective cell types. Upon uptake into proliferating cells,MTABGNRs were diluted equally and relatively rapidly into daughter cells; however, in nondividing/senescent cells the loss ofMTABGNRs was gradual and very modest, attributable mainly to exocytosis. ExocytosedMTABGNRs can again be internalized. These findings broaden our knowledge about cellular uptake of gold nanorods, a crucial prerequisite for future successful engineering of nanoparticles for biomedical applications such as photothermal cancer therapy or elimination of senescent cells as part of the emerging rejuvenation approach.
Department of Genome Integrity, Institute of Molecular Genetics of the CAS, v.v.i. , CZ-142 20 Prague 4, Czech Republic.
Monika
M
Zarska
Department of Genome Integrity, Institute of Molecular Genetics of the CAS, v.v.i. , CZ-142 20 Prague 4, Czech Republic.|Department of Physical Electronics, Faculty of Nuclear Sciences and Physical Engineering, Czech Technical University in Prague , CZ-115 19 Prague 1, Czech Republic.
Filip
F
Novotny
Department of Genome Integrity, Institute of Molecular Genetics of the CAS, v.v.i. , CZ-142 20 Prague 4, Czech Republic.|Department of Physical Electronics, Faculty of Nuclear Sciences and Physical Engineering, Czech Technical University in Prague , CZ-115 19 Prague 1, Czech Republic.
Filip
F
Havel
Department of Genome Integrity, Institute of Molecular Genetics of the CAS, v.v.i. , CZ-142 20 Prague 4, Czech Republic.
Michal
M
Sramek
Laboratory of Mutagenesis and Carcinogenesis, Cancer Research Institute BMC, Slovak Academy of Sciences , 945 05 Bratislava, Slovakia.
Andrea
A
Babelova
Laboratory of Molecular Structure Characterization, Institute of Microbiology of the CAS, v.v.i. , CZ-142 20 Prague 4, Czech Republic.
Oldrich
O
Benada
Department of Chemistry, Faculty of Science, University of Hradec Kralove , 500 03 Hradec Kralove, Czech Republic.|Biomedical Research Center, University Hospital , CZ-500 05 Hradec Kralove, Czech Republic.
Michal
M
Novotny
Department of Chemistry, Faculty of Science, University of Hradec Kralove , 500 03 Hradec Kralove, Czech Republic.
Hilal
H
Saran
Department of Chemistry, Faculty of Science, University of Hradec Kralove , 500 03 Hradec Kralove, Czech Republic.|Biomedical Research Center, University Hospital , CZ-500 05 Hradec Kralove, Czech Republic.
Kamil
K
Kuca
Department of Chemistry, Faculty of Science, University of Hradec Kralove , 500 03 Hradec Kralove, Czech Republic.|Biomedical Research Center, University Hospital , CZ-500 05 Hradec Kralove, Czech Republic.
Kamil
K
Musilek
Department of Genome Integrity, Institute of Molecular Genetics of the CAS, v.v.i. , CZ-142 20 Prague 4, Czech Republic.
Zuzana
Z
Hvezdova
Department of Genome Integrity, Institute of Molecular Genetics of the CAS, v.v.i. , CZ-142 20 Prague 4, Czech Republic.
Rastislav
R
Dzijak
Department of Genome Integrity, Institute of Molecular Genetics of the CAS, v.v.i. , CZ-142 20 Prague 4, Czech Republic.
Marketa
M
Vancurova
Department of Genome Integrity, Institute of Molecular Genetics of the CAS, v.v.i. , CZ-142 20 Prague 4, Czech Republic.
Katerina
K
Krejcikova
Department of Genome Integrity, Institute of Molecular Genetics of the CAS, v.v.i. , CZ-142 20 Prague 4, Czech Republic.
Blanka
B
Gabajova
Department of Genome Integrity, Institute of Molecular Genetics of the CAS, v.v.i. , CZ-142 20 Prague 4, Czech Republic.
Hana
H
Hanzlikova
Department of Genome Integrity, Institute of Molecular Genetics of the CAS, v.v.i. , CZ-142 20 Prague 4, Czech Republic.
Lenka
L
Kyjacova
Department of Genome Integrity, Institute of Molecular Genetics of the CAS, v.v.i. , CZ-142 20 Prague 4, Czech Republic.|Genome Integrity Unit, Danish Cancer Society Research Center , DK-2100 Copenhagen, Denmark.|Department of Medical Biochemistry and Biophysics, Science For Life Laboratory, Division of Translational Medicine and Chemical Biology, Karolinska Institute , 17121 Solna, Sweden.
Jiri
J
Bartek
Department of Genome Integrity, Institute of Molecular Genetics of the CAS, v.v.i. , CZ-142 20 Prague 4, Czech Republic.|Department of Physical Electronics, Faculty of Nuclear Sciences and Physical Engineering, Czech Technical University in Prague , CZ-115 19 Prague 1, Czech Republic.
Jan
J
Proska
Department of Genome Integrity, Institute of Molecular Genetics of the CAS, v.v.i. , CZ-142 20 Prague 4, Czech Republic.
Zdenek
Z
Hodny
D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D049109:Cell Proliferation / Q000187:drug effects; D060326:Chemistry Techniques, Synthetic; D003470:Culture Media; D004355:Drug Stability; D004705:Endocytosis / Q000187:drug effects / Q000502:physiology; D005089:Exocytosis / Q000187:drug effects / Q000502:physiology; D005434:Flow Cytometry; D006046:Gold / Q000737:chemistry* / Q000493:pharmacokinetics*; D006801:Humans; D008247:Lysosomes / Q000187:drug effects; D018613:Microscopy, Confocal; D008855:Microscopy, Electron, Scanning; D043942:Nanotubes / Q000032:analysis / Q000737:chemistry*; D011107:Polylysine / Q000737:chemistry* / Q000493:pharmacokinetics*; D011509:Proteoglycans / Q000737:chemistry / Q000378:metabolism; D000644:Quaternary Ammonium Compounds / Q000138:chemical synthesis / Q000737:chemistry*; D013438:Sulfhydryl Compounds / Q000737:chemistry*
C000620117:(16-mercaptohexadecyl)trimethylammonium bromide; D003470:Culture Media; D011509:Proteoglycans; D000644:Quaternary Ammonium Compounds; D013438:Sulfhydryl Compounds; C089532:cationic colloidal gold; D011107:Polylysine; D006046:Gold