title: Statistical analysis of gold nanoparticle-induced oxidative stress and apoptosis in myoblast (C2C12) cells.
abstract: Nanoscale gold particles (Au-NPs) with a diameter below 20nm are notably important candidates for various important applications because of their extraordinary quantum size effects. Their high surface area-to-volume ratio facilitates their very high reactivities; therefore, they can be utilised in different ways in biomedical applications. For example, these nanoparticles can penetrate into cells and bind with proteins or DNA and are therefore potential nanostructures employed for sensing and detecting various biological identities. In the present work, we synthesised Au-NPs via a colloidal process using chloroauric acid (HAuCl4·4H2O) and trisodium citrate dihydrate (N3C6H5O7) as a reducing agent. The shape evolution and the structural properties of these NPs were investigated in detail using TEM and high resolution HR-TEM investigations. Different doses of Au NPs have been applied to treat C2C12 myoblast cells in a 24-h incubation period, and a dose-dependent study has also been performed. The cells were cultivated in DMEM with FBS and antibiotics (strepto-penicillin) at 37°C in a 5% humidified environment of CO2 and 95% air. Cell viability analysis using MTT assays revealed that increased concentration of Au NPs (100-1000 ng/mL) resulted in a decreased density of cells. The amount of reactive oxygen species (ROS) in C2C12 cells analysed with Au-NPs (in a dose-dependent manner), and the RT-PCR data demonstrated the up-regulation of caspase-3 and caspase-7 genes in C2C12 cells after treatment with Au-NPs. These results have been confirmed by detailed confocal microscopy (CLSM) studies. In addition, the quantitative analysis of the Au-NPs was also confirmed by statistical analytical parameters, such as precision, accuracy, linearity, limits of detection (LOD) and limit of quantitation (LOQ), quantitative recoveries and relative standard deviation (RSD), and the analyses again exhibited a significant and large effect of Au NPs on C2C12 cells.
authors.affiliation: A.R.Al-Jeraisy, Chair for DNA Research, Department of Zoology, College of Science, King Saud University, Riyadh 11451, Saudi Arabia. Electronic address: rwahab05@gmail.com.
authors.forename: Rizwan
authors.initials: R
authors.lastname: Wahab
authors.affiliation: A.R.Al-Jeraisy, Chair for DNA Research, Department of Zoology, College of Science, King Saud University, Riyadh 11451, Saudi Arabia.
authors.forename: Sourabh
authors.initials: S
authors.lastname: Dwivedi
authors.affiliation: Department of Chemistry, Aligarh Muslim University, Aligarh 202002, India.
authors.forename: Farheen
authors.initials: F
authors.lastname: Khan
authors.affiliation: Functional Nanomaterials, Institute for Materials Science, University of Kiel, Kaiser Str. 2, 24143 Kiel, Germany.
authors.forename: Yogendra K
authors.initials: YK
authors.lastname: Mishra
authors.affiliation: Department of Animal Sciences and Biotechnology, Chonbuk National University Jeonju, Jeollabuk-do 561-756, Republic of Korea.
authors.forename: I H
authors.initials: IH
authors.lastname: Hwang
authors.affiliation: Energy Materials and Surface Science Laboratory, Solar Energy Research Centre, School of Chemical Engineering, Chonbuk National University, Jeonju 561-756, Republic of Korea.
authors.forename: Hyung-Shik
authors.initials: HS
authors.lastname: Shin
authors.affiliation: Department of Agricultural Microbiology, Faculty of Agricultural Sciences, Aligarh Muslim University, Aligarh 202002, India.
authors.forename: Javed
authors.initials: J
authors.lastname: Musarrat
authors.affiliation: A.R.Al-Jeraisy, Chair for DNA Research, Department of Zoology, College of Science, King Saud University, Riyadh 11451, Saudi Arabia.
authors.forename: Abdulaziz A
authors.initials: AA
authors.lastname: Al-Khedhairy
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D053148:Caspase 3 / Q000378:metabolism; D053179:Caspase 7 / Q000378:metabolism; D006046:Gold / Q000737:chemistry*; D053768:Metal Nanoparticles / Q000737:chemistry*; D051379:Mice; D032446:Myoblasts / Q000166:cytology* / Q000187:drug effects*; D018384:Oxidative Stress / Q000187:drug effects*; D017382:Reactive Oxygen Species / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D017382:Reactive Oxygen Species; D006046:Gold; D053148:Caspase 3; D053179:Caspase 7
keywords: Apoptosis; C2C12 cells; Colloidal process; Gold nanoparticles; MTT assay