A Flow Cytometry‐based Method for the Screening of Nanomaterial‐induced Reactive Oxygen Species Production in Leukocytes Subpopulations in Whole Blood

Abstract

To date, the use and translation of nanomedicine from the laboratory to the clinic has been relatively slow. Among other issues, one of the reasons for this tardiness is the lack of the availability of quick and reliable toxicity tools for the screening of nanomaterials (NMs). In this investigation, we apply a flow cytometry-based method for the detection of nanomaterialinduced oxidative stress by measurement of reactive oxygen species production in specific leukocyte subpopulations in human whole blood. The screening of a panel of relevant nanomedical-associated materials (liposomes, silica, iron oxide and functionalized single-walled carbon nanotubes) demonstrated that only the carbon nanotubes induced oxidative stress in human circulating leukocytes. In summary, we apply and corroborate a flow cytometry-based method for the simple and effective measurement of NM-induced oxidative stress in human blood subpopulations after realistic and relevant exposure scenarios which is extremely useful in future toxicological applications.The rapid development of nanotechnology goes hand in hand with concerns about the potential adverse effects of nanomaterial (NM) exposure on human health and the environment. In addition, the apparent knowledge gaps in our understanding of the interaction between NMs and cells, their transformation and their eventual fate in different biological systems mean that hazard assessment is very challenging.Nanomedicine is defined as the application and utilization of nanotechnology to medicine and human health, in particular with regard to the diagnosis and treatment of disease. Over the last two decades, a wide range of NMs with varying size, shape, charge and other physicochemical characteristics are tailored to determine specific function, resulting in desirable optical, electronic, magnetic and biological attributes. The current nanomedicine applications mainly include vehicles for targeted drug delivery, NM platforms for biomedical imaging and therapeutic applications for treating clinical diseases [1,2]. Despite these promising medical applications, the safety concern for NMs is a key determinant factor in the assessment of their clinical potential. Therefore, it is imperative to implement risk reduction strategies for all materials proposed for medical applications. The same physicochemical characteristics that make NMs desirable for medical applications might contribute to their potential adverse effects -particle size and surface properties can largely influence the bioavailability, transport, biotransformation, cellular uptake and toxicity of the material in question [3,4].Iron oxide nanoparticles (NPs) can exhibit a unique form of superparamagnetism which is highly useful and desirable in biomedicine for several medical applications, principally relating to imaging and drug delivery to the central nervous system and various tumours [5,6]. This property is believed to be size-dependent and only occurs at around 10-20 nm [7]. Iron oxide is one of the few NMs with current clin...

Quantitative Flow Cytometric Evaluation of Oxidative Stress and Mitochondrial Impairment in RAW 264.7 Macrophages after Exposure to Pristine, Acid Functionalized, or Annealed Carbon Nanotubes

“…Cellular debris can act as artefactual objects, leading to the poor performances of vital probes, and it may distort the quantitation of toxicological outcomes [59][60][61]. In this context, flow cytometry (FCM) appears as a powerful alternative methodology to perform bio-toxicological assays avoiding as much as possible any residual presence of cell debris and MWCNTs aggregates that could interfere, and at the same time discriminating cell subpopulations of interest [12,25,[62][63][64][65]. We developed here a quantitative multiparametric FCM approach to investigate early alterations such as oxidative stress generation and mitochondrial potential alteration, and late events such as cytotoxicity (cell death rate) and genotoxicity (apoptotic DNA fragmentation and chromatin decondensation).…”

Divergent Responses of Hydrophilic CdSe and CdSe@CdS Core–Shell Nanocrystals in Apoptosis and In Vitro Cancer Cell Imaging: A Comparative Analysis

“…Intracellular ROS induced by CdSe and CdSe@CdS NCs were detected by using 2′,7′-Dichlorofluorescein-diacetate (DCFH-DA) staining following the protocols of Kermanizadeh et al [ 35 ]. The fluorescence intensity was measured using 485 excitation and 520 nm emission filters using a fluorimeter (RF-5301 PC Shimadzu spectrofluorometer, Nakagyo-ku, Kyoto, Japan).…”

“…EPR measurements of free radicals were carried out in a Bruker EMX Micro X spectrometer according to the modified protocol of Kaur et al [ 35 ]. The treated cells were suspended in 100 mM DMPO.…”

A Review of the Current State of Nanomedicines for Targeting and Treatment of Cancers: Achievements and Future Challenges

“…In recent years, great progress has been made in the synthesis of a variety of materials that can be used as nanovehicles (carbons, synthetic polymers, micelles, liposomes/vesicles, drug–polymer polysaccharide conjugates to name a few). [ 7–10 ] Currently, commercial nanomedicines are predominantly generated as nanocarriers for therapeutics or stabilized solid drug NMs. As an example, NM surfaces can be chemically modified to influence site‐specific drug delivery.…”