Frontiers in Bioengineering and Biotechnology, Journal Year: 2025, Volume and Issue: 13

Published: April 30, 2025

The safe-by-design of engineered nanoparticles (NPs) for any application requires a detailed understanding how the particles interact with single cells. Most studies are based on two-dimensional, uniformly dense cell cultures, which do not represent diverse and inhomogeneous environments found in situ. In-vitro models that accurately tissue complexity, including realistic densities, essential to increase predictive accuracy cell-NP interactions. This study uses bioprinted gradient model examine relation between density NP uptake one dish. A549 lung epithelial gradients within inserts were produced bioprinter by modulating inter-droplet distances. After two days culture, cells exposed Cy5-labeled silica NPs (SiO2 NPs, ∼112 nm, 20 μg/mL) up 48 h. Confocal fluorescence microscopy 3D image analysis used quantify uptake, surface area, volume. relationship other parameters was then investigated statistically. Bioprinting enabled creation reproducible linear gradients, allowing controlled modeling variations while preserving viability throughout experiment. Increasing distances, from 0.1 mm 0.6 mm, achieve decreasing densities. SiO2 per around 50% higher low-density regions compared high-density areas across all time points, i.e., 6, 24, h post-exposure. inverse correlated greater average area lower-density regions, differences proliferation rates at varying densities did significantly impact uptake. is enhanced lower mainly due increased available revealing potential interaction tissues present variability. Our drop-on-demand bioprinting successfully supports implementation in-vitro their relevance as new approach methodologies (NAMs) next-generation risk assessment strategies.

Language: Английский