Fingerprinting Metabolic Activity and Tissue Integrity of 3D Lung Cancer Spheroids under Gold Nanowire Treatment.

Ali Hamad Abd Kelkawi,Abdollah Allahverdi,Hossein Naderi-Manesh,Hadi Hashemzadeh,Peter Ertl,Mario Rothbauer

doi:10.3390/cells11030478

Ali Hamad Abd Kelkawi, Abdollah Allahverdi + Show 4 more

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https://doi.org/10.3390/cells11030478

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Abstract

Inadequacy of most animal models for drug efficacy assessments has led to the development of improved in vitro models capable of mimicking in vivo exposure scenarios. Among others, 3D multicellular spheroid technology is considered to be one of the promising alternatives in the pharmaceutical drug discovery process. In addition to its physiological relevance, this method fulfills high-throughput and low-cost requirements for preclinical cell-based assays. Despite the increasing applications of spheroid technology in pharmaceutical screening, its application, in nanotoxicity testing is still in its infancy due to the limited penetration and uptake rates into 3D-cell assemblies. To gain a better understanding of gold nanowires (AuNWs) interactions with 3D spheroids, a comparative study of 2D monolayer cultures and 3D multicellular spheroids was conducted using two lung cancer cell lines (A549 and PC9). Cell apoptosis (live/dead assay), metabolic activity, and spheroid integrity were evaluated following exposure to AuNWs at different dose-time manners. Results revealed a distinct different cellular response between 2D and 3D cell cultures during AuNWs treatment including metabolic rates, cell viability, dose–response curves and, uptake rates. Our data also highlighted further need for more physiologically relevant tissue models to investigate in depth nanomaterial–biology interactions. It is important to note that higher concentrations of AuNWs with lower exposure times and lower concentrations of AuNWs with higher exposure times of 3 days resulted in the loss of spheroid integrity by disrupting cell–cell contacts. These findings could help to increase the understanding of AuNWs-induced toxicity on tissue levels and also contribute to the establishment of new analytical approaches for toxicological and drug screening studies.

Highlights

The global trend towards three-dimensional (3D) in vitro cell cultures systems is partially driven by the fundamental biological changes, such as gene/protein profiles and cell signaling pathways, that take place when moving from classical 2D cultures to 3D cell assemblies [1,2,3]
It was noticed that the 3D spheroid integrity of lung cancer cells were more sensitive to the a better understanding of gold nanowires (AuNWs) toxicity than 2D cell culture treatments
One of the most important factors influencing the results of 2D and 3D spheroid changes is known as the ECM of solid tumors

Summary

Introduction

The global trend towards three-dimensional (3D) in vitro cell cultures systems is partially driven by the fundamental biological changes, such as gene/protein profiles and cell signaling pathways, that take place when moving from classical 2D cultures to 3D cell assemblies [1,2,3]. Oxygen (O2), pH and nutrient gradients inside of a MCTS are similar to the in vivo solid tumors [9]. These 3D cell culture models are introduced into industrial processes as powerful systems with massive translational potential including stem cell research, cancer research, drug discovery, cell-based biosensors, and toxicological effects studies [10,11,12,13,14,15]. Recapitulation of in vivo morphology, mimicry of tissue architecture, good cell connectivity such as cell–cell and cell–matrix interactions and gene and protein expression profile similarity with physiological microenvironment, provide 3D cell culture models with a predictive perspective on physiological tissue behavior under drug treatment.

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