Statistical analysis of gold nanoparticle-induced oxidative stress and apoptosis in myoblast (C2C12) cells

Rizwan Wahab; Sourabh Dwivedi; Farheen Khan; Yogendra K Mishra; I H Hwang; Hyung-Shik Shin; Javed Musarrat; Abdulaziz A Al-Khedhairy

doi:10.1016/j.colsurfb.2014.10.012

Statistical analysis of gold nanoparticle-induced oxidative stress and apoptosis in myoblast (C2C12) cells

Colloids Surf B Biointerfaces. 2014 Nov 1:123:664-72. doi: 10.1016/j.colsurfb.2014.10.012. Epub 2014 Oct 12.

Authors

Rizwan Wahab¹, Sourabh Dwivedi², Farheen Khan³, Yogendra K Mishra⁴, I H Hwang⁵, Hyung-Shik Shin⁶, Javed Musarrat⁷, Abdulaziz A Al-Khedhairy²

Affiliations

¹ 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.
² A.R.Al-Jeraisy, Chair for DNA Research, Department of Zoology, College of Science, King Saud University, Riyadh 11451, Saudi Arabia.
³ Department of Chemistry, Aligarh Muslim University, Aligarh 202002, India.
⁴ Functional Nanomaterials, Institute for Materials Science, University of Kiel, Kaiser Str. 2, 24143 Kiel, Germany.
⁵ Department of Animal Sciences and Biotechnology, Chonbuk National University Jeonju, Jeollabuk-do 561-756, Republic of Korea.
⁶ Energy Materials and Surface Science Laboratory, Solar Energy Research Centre, School of Chemical Engineering, Chonbuk National University, Jeonju 561-756, Republic of Korea.
⁷ Department of Agricultural Microbiology, Faculty of Agricultural Sciences, Aligarh Muslim University, Aligarh 202002, India.

PMID: 25456994
DOI: 10.1016/j.colsurfb.2014.10.012

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.

Keywords: Apoptosis; C2C12 cells; Colloidal process; Gold nanoparticles; MTT assay.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Apoptosis / drug effects
Caspase 3 / metabolism
Caspase 7 / metabolism
Gold / chemistry*
Metal Nanoparticles / chemistry*
Mice
Myoblasts / cytology*
Myoblasts / drug effects*
Oxidative Stress / drug effects*
Reactive Oxygen Species / metabolism
Reverse Transcriptase Polymerase Chain Reaction

Substances

Reactive Oxygen Species
Gold
Caspase 3
Caspase 7