Abstract
Most cell culture models are static, but the cellular microenvironment in the body is dynamic. Here, we established a microfluidic-based in vitro model of human bronchial epithelial cells in which cells are stationary, but nutrient supply is dynamic, and we used this system to evaluate cellular uptake of nanoparticles. The cells were maintained in fetal calf serum-free and bovine pituitary extract-free cell culture medium. BEAS-2B, an immortalized, non-tumorigenic human cell line, was used as a model and the cells were grown in a chip within a microfluidic device and were briefly infused with amorphous silica (SiO2) nanoparticles or polystyrene (PS) nanoparticles of similar primary sizes but with different densities. For comparison, tests were also performed using static, multi-well cultures. Cellular uptake of the fluorescently labeled particles was investigated by flow cytometry and confocal microscopy. Exposure under dynamic culture conditions resulted in higher cellular uptake of the PS nanoparticles when compared to static conditions, while uptake of SiO2 nanoparticles was similar in both settings. The present study has shown that it is feasible to grow human lung cells under completely animal-free conditions using a microfluidic-based device, and we have also found that cellular uptake of PS nanoparticles aka nanoplastics is highly dependent on culture conditions. Hence, traditional cell cultures may not accurately reflect the uptake of low-density particles, potentially leading to an underestimation of their cellular impact.
Highlights
Experts in the field have argued that “nanotoxicology is currently at a crossroads and faces a number of obstacles and technical limitations not associated with traditional toxicology” (Hussain et al, 2015)
To allow for the comparison between the two set-ups, we first determined the cell density, cell viability, and cell morphology of the BEAS-2B cells cultured in 24-well cell culture plates vs. in the microfluidic device (Supplementary Table S1, Figure S1)
The BEAS-2B cells were maintained and exposed under completely animal-free conditions in this study meaning that no animal-derived products were applied
Summary
Experts in the field have argued that “nanotoxicology is currently at a crossroads and faces a number of obstacles and technical limitations not associated with traditional toxicology” (Hussain et al, 2015). The field of nanotoxicology still relies heavily on assays and methods developed for the testing of traditional chemicals, and the development of relevant and robust assays amenable to highthroughput screening of nanomaterials represents an important priority (Li et al, 2018; Fadeel, 2019). There is a strong consensus that faster and animal-free approaches for safety assessment of Microfluidic-Based Lung Cell Model chemicals as well as engineered nanomaterials are needed (Kohl et al, 2021). More recent developments include the design of multiorgan-on-a-chip devices in an attempt to capture the crosstalk between different cell types (Ashammakhi et al, 2020). Recent attempts have been made to grow tumor spheroids in a microfluidic device to more accurately model and determine NP uptake (Zhuang et al, 2019)
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