title: Enhanced intracellular translocation and biodistribution of gold nanoparticles functionalized with a cell-penetrating peptide (VG-21) from vesicular stomatitis virus.
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-2 cells. Cytokine response to fGNPs was studied in vitro and in vivo. 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-2 cells 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-7 cells, in endocytotic vesicles and nuclei. Cytokine ELISA showed that mouse J774 cells 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 35 ng/mg tissue and 20 ng/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.
authors.affiliation: Center for NanoBiotechnology Research and Department of Biological Sciences, Alabama State University, 1627 Hall Street, Montgomery, AL 36101, USA.
authors.forename: Pooja Munnilal
authors.initials: PM
authors.lastname: Tiwari
authors.affiliation: Center for NanoBiotechnology Research and Department of Biological Sciences, Alabama State University, 1627 Hall Street, Montgomery, AL 36101, USA; Faculty of Engineering, Bioengineering Department, Celal Bayar University, 45140 Muradiye, Manisa, Turkey.
authors.forename: Erdal
authors.initials: E
authors.lastname: Eroglu
authors.affiliation: Center for NanoBiotechnology Research and Department of Biological Sciences, Alabama State University, 1627 Hall Street, Montgomery, AL 36101, USA.
authors.forename: Swapnil Subhash
authors.initials: SS
authors.lastname: Bawage
authors.affiliation: Center for NanoBiotechnology Research and Department of Biological Sciences, Alabama State University, 1627 Hall Street, Montgomery, AL 36101, USA.
authors.forename: Komal
authors.initials: K
authors.lastname: Vig
authors.affiliation: Auburn University Research Instrumentation Facility, Harrison School of Pharmacy, 32 Rouse Life Sciences Building, Auburn University, Auburn, AL 36849, USA.
authors.forename: Michael E
authors.initials: ME
authors.lastname: Miller
authors.affiliation: Center for NanoBiotechnology Research and Department of Biological Sciences, Alabama State University, 1627 Hall Street, Montgomery, AL 36101, USA.
authors.forename: Shreekumar
authors.initials: S
authors.lastname: Pillai
authors.affiliation: Center for NanoBiotechnology Research and Department of Biological Sciences, Alabama State University, 1627 Hall Street, Montgomery, AL 36101, USA.
authors.forename: Vida A
authors.initials: VA
authors.lastname: Dennis
authors.affiliation: Center for NanoBiotechnology Research and Department of Biological Sciences, Alabama State University, 1627 Hall Street, Montgomery, AL 36101, USA. Electronic address: ssingh@alasu.edu.
authors.forename: Shree Ram
authors.initials: SR
authors.lastname: Singh
mesh_terms: D000818:Animals; D019556:COS Cells; D002470:Cell Survival / Q000187:drug effects; D057846:Cell-Penetrating Peptides / Q000493:pharmacokinetics*; D002522:Chlorocebus aethiops; D005260:Female; D006046:Gold / Q000737:chemistry / Q000493:pharmacokinetics*; D006367:HeLa Cells; D006801:Humans; D015850:Interleukin-6 / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000493:pharmacokinetics; D053768:Metal Nanoparticles / Q000737:chemistry*; D051379:Mice; D008807:Mice, Inbred BALB C; D046529:Microscopy, Electron, Transmission; D014018:Tissue Distribution; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism; D014709:Vero Cells; D018116:Vesiculovirus / Q000378:metabolism; D014759:Viral Envelope Proteins / Q000493:pharmacokinetics
chemical_list: D057846:Cell-Penetrating Peptides; C028391:G protein, vesicular stomatitis virus; D015850:Interleukin-6; D008562:Membrane Glycoproteins; D014409:Tumor Necrosis Factor-alpha; D014759:Viral Envelope Proteins; D006046:Gold
keywords: Cell-penetrating peptide; Drug delivery; Functionalized gold nanoparticles; Vesicular stomatitis virus