Comparison of cytotoxicity and membrane efflux pump inhibition in HepG2 cells induced by single-walled carbon nanotubes with different length and functional groups

Zhuoyan Shen,Su Liu,Jialu Wu,Wei Jiang,Habiba Nurmamat,Bing Wu,Yue Yu

doi:10.1038/s41598-019-43900-5

Abstract

Environmental risk of single-walled carbon nanotubes (SWCNTs) is receiving increasing attentions owing to their wide study and application. However, little is known on the influence of length and functional groups on SWCNT cytotoxicity. In this study, six types of SWCNTs with different functional groups (pristine, carboxyl group and hydroxyl group) and lengths (1–3 μm and 5–30 μm) were chosen. Cytotoxicities in human hepatoma HepG2 cells induced by these SWCNTs were compared based on cell viability, oxidative stress, plasma membrane fluidity and ABC transporter activity assays. Results showed that all the SWCNTs decreased cell viability of HepG2, increased intracellular reactive oxygen species (ROS) level, and damaged plasma membrane in a concentration-dependent manner. Long SWCNTs had stronger cytotoxic effects than short SWCNTs, which might be due to weaker aggregation for the long SWCNTs. Functionalization changed the toxic effects of the SWCNTs, and different influence was found between long SWCNTs and short SWCNTs. Moreover, the six types of SWCNTs at low concentrations changed plasma membrane fluidity, inhibited transmembrane ABC transporter (efflux pump) activity, and acted as chemosensitizer to improve the sensitivity of cells to arsenic, indicating the chemosensitive effect should be considered as toxic endpoint of SWCNTs. Comparison of different toxic endpoints among the six types of SWCNTs showed that short hydroxyl-SWCNT might be safer than other SWCNTs. This study provides insights into toxicities of SWCNTs, which is of great value for the risk assessment and application of SWCNTs.

Highlights

Nanomaterials have been widely applied in many fields, including physical, chemical, mechanical, electrical and biomedical areas[1,2,3]
Results of dynamic light scattering (DLS) analyses showed that the long single-walled carbon nanotubes (SWCNTs) had larger hydrodynamic size in cell culture media than the short SWCNTs (Table S1)
Concentration of SWCNTs used in DLS measurement was 5 mg/L, which was in the range of exposure concentrations for cytotoxicity assays

Summary

Introduction

Nanomaterials have been widely applied in many fields, including physical, chemical, mechanical, electrical and biomedical areas[1,2,3]. Compared to MWCNTs, fewer information on cytotoxicity of SWCNTs are available, and the available data are always contradictory. Some studies showed that SWCNTs exposure induced oxidative stress and inflammation[13,14,15,16,17]. Horie et al.[26] thought the stability of SWCNTs suspension had more influence on cytotoxicity compared to their length. Results show that the length and functionalization can change the cytotoxicity of SWCNTs. it is necessary to systematically determine the influence of length and functionalization on cytotoxic effects induced by SWCNTs. Objective of this study is to compare cytotoxicity of SWCNTs with different lengths and functional groups in human hepatoma HepG2 cells. This study could provide meaningful information for risk assessment and applications of SWCNTs with different lengths and functional groups

Methods

Results

Discussion

Conclusion