title: Surface charge of gold nanoparticles mediates mechanism of toxicity.
abstract: Recently gold nanoparticles (Au NPs) have shown promising biological and military applications due to their unique electronic and optical properties. However, little is known about their biocompatibility in the event that they come into contact with a biological system. In the present study, we have investigated whether modulating the surface charge of 1.5 nm Au NPs induced changes in cellular morphology, mitochondrial function, mitochondrial membrane potential (MMP), intracellular calcium levels, DNA damage-related gene expression, and of p53 and caspase-3 expression levels after exposure in a human keratinocyte cell line (HaCaT). The evaluation of three different Au NPs (positively charged, neutral, and negatively charged) showed that cell morphology was disrupted by all three NPs and that they demonstrated a dose-dependent toxicity; the charged Au NPs displayed toxicity as low as 10 µg ml(-1) and the neutral at 25 µg ml(-1). Furthermore, there was significant mitochondrial stress (decreases in MMP and intracellular Ca2+ levels) following exposure to the charged Au NPs, but not the neutral Au NPs. In addition to the differences observed in the MMP and Ca2+ levels, up or down regulation of DNA damage related gene expression suggested a differential cell death mechanism based on whether or not the Au NPs were charged or neutral. Additionally, increased nuclear localization of p53 and caspase-3 expression was observed in cells exposed to the charged Au NPs, while the neutral Au NPs caused an increase in both nuclear and cytoplasmic p53 expression. In conclusion, these results indicate that surface charge is a major determinant of how Au NPs impact cellular processes, with the charged NPs inducing cell death through apoptosis and neutral NPs leading to necrosis.
authors.affiliation: Applied Biotechnology Branch, Human Effectiveness Directorate, 711 Human Performance Wing, Air Force Research Laboratory, Area B, R Street, BLDG 837, Wright-Patterson AFB, OH 45433-5707, USA.
authors.forename: Nicole M
authors.initials: NM
authors.lastname: Schaeublin
authors.forename: Laura K
authors.initials: LK
authors.lastname: Braydich-Stolle
authors.forename: Amanda M
authors.initials: AM
authors.lastname: Schrand
authors.forename: John M
authors.initials: JM
authors.lastname: Miller
authors.forename: Jim
authors.initials: J
authors.lastname: Hutchison
authors.forename: John J
authors.initials: JJ
authors.lastname: Schlager
authors.forename: Saber M
authors.initials: SM
authors.lastname: Hussain
mesh_terms: D017209:Apoptosis; D002118:Calcium / Q000378:metabolism; D053148:Caspase 3 / Q000378:metabolism; D002460:Cell Line; D004249:DNA Damage; D006046:Gold / Q000737:chemistry*; D006801:Humans; D015603:Keratinocytes / Q000166:cytology; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D053768:Metal Nanoparticles / Q000737:chemistry* / Q000633:toxicity; D008928:Mitochondria / Q000378:metabolism / Q000502:physiology; D017382:Reactive Oxygen Species / Q000378:metabolism; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism
chemical_list: D017382:Reactive Oxygen Species; D016159:Tumor Suppressor Protein p53; D006046:Gold; D053148:Caspase 3; D002118:Calcium