Enhanced intracellular translocation and biodistribution of gold nanoparticles functionalized with a cell-penetrating peptide (VG-21) from vesicular stomatitis virus
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Date
2014
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Abstract
Reduced toxicity and ease of modification make gold nanoparticles (GNPs) suitable for targeted delivery, bioimaging and theranostics by conjugating cell-penetrating peptides (CPPs). This study presents the biodistribution and enhanced intracellular uptake of GNPs functionalized with VG-21, a CPP derived from vesicular stomatitis virus glycoprotein (G). Cell penetrating efficiency of VG-21 was demonstrated using CellPPD web server, conjugated to GNPs and were characterized using, UV-visible and FTIR spectroscopy, transmission electron microscopy, dynamic light scattering and zeta potential. Uptake of VG-21 functionalized GNPs (fGNPs) was tested in eukaryotic cell lines, HEp-2, HeLa, Vero and Cos-7, using flow cytometry, fluorescence and transmission electron microscopy (TEM), and inductively coupled plasmon optical emission spectroscopy (ICP-OES). The effects of nanoparticles on stress and toxicity related genes were studied in HEp-2cells. Cytokine response to fGNPs was studied invitro and invivo. Biodistribution of nanoparticles was studied in BALB/c mice using TEM and ICP-OES. VG-21, GNPs and fGNPs had little to no effect on cell viability. Upon exposure to fGNPs, HEp-2cells revealed minimal down regulation of stress response genes. fGNPs displayed higher uptake than GNPs in all cell lines with highest internalization by HEp-2, HeLa and Cos-7cells, in endocytotic vesicles and nuclei. Cytokine ELISA showed that mouse J774cells exposed to fGNPs produced less IL-6 than did GNP-treated macrophage cells, whereas TNF-α levels were low in both treatment groups. Biodistribution studies in BALB/c mice revealed higher accumulation of fGNPs than GNPs in the liver and spleen. Histopathological analyses showed that fGNP-treated mice accumulated 35ng/mg tissue and 20ng/mg tissue gold in spleen and liver respectively, without any adverse effects. Likewise, serum cytokines were low in both GNP- and fGNP-treated mice. Thus, VG-21-conjugated GNPs have enhanced cellular internalization and are suitable for various biomedical applications as nano-conjugates. © 2014 The Authors.
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Animals , Cell Survival , Cell-Penetrating Peptides , Cercopithecus aethiops , COS Cells , Female , Gold , HeLa Cells , Humans , Interleukin-6 , Membrane Glycoproteins , Metal Nanoparticles , Mice , Mice, Inbred BALB C , Microscopy, Electron, Transmission , Tissue Distribution , Tumor Necrosis Factor-alpha , Vero Cells , Vesiculovirus , Viral Envelope Proteins , Eukaryota , Mus , Vesicular stomatitis virus , Cell culture , Cytology , Drug delivery , Electromagnetic induction , Electron microscopy , Emission spectroscopy , Fiber optic sensors , Fourier transform infrared spectroscopy , Genes , Gold , Light scattering , Mammals , Medical applications , Metal nanoparticles , Mobile security , Nanoparticles , Optical emission spectroscopy , Peptides , Tissue , Tissue engineering , Toxicity , Transmission electron microscopy , Viruses , cell penetrating peptide , cytokine , drug vehicle , gold nanoparticle , interleukin 6 , nanoconjugate , nanoparticle , protein VG 21 , protein VG 21 gold nanoparticle conjugate , tumor necrosis factor alpha , unclassified drug , cell penetrating peptide , G protein, vesicular stomatitis virus , gold , interleukin 6 , membrane protein , metal nanoparticle , tumor necrosis factor alpha , virus envelope protein , Cell penetrating peptides (CPPs) , Cell-penetrating peptide , Cellular internalization , Functionalized gold nanoparticles , Gold nanoparticles (GNPs) , Histopathological analysis , Intracellular translocation , Vesicular stomatitis virus , animal cell , animal experiment , animal tissue , Article , cell nucleus , cell viability , controlled study , COS 7 cell line , cytokine production , drug accumulation , drug conjugation , drug delivery system , drug distribution , drug effect , drug transport , endosome , enzyme linked immunosorbent assay , flow cytometry , fluorescence microscopy , gene expression , HeLa cell line , HEp 2 cell line , histopathology , human , human cell , in vitro study , in vivo study , inductively coupled plasmon optical emission spectroscopy , infrared spectroscopy , internalization , light scattering , liver , macrophage , mass spectrometry , mouse , nonhuman , oxidative stress , priority journal , spleen , tissue distribution , transmission electron microscopy , ultraviolet spectroscopy , Vero cell line , Vesiculovirus , zeta potential , animal , Bagg albino mouse , cell survival , chemistry , Chlorocebus aethiops , COS 1 cell line , drug effects , female , metabolism , Vesiculovirus , Cells