A novel workflow for generating representative and cell-culture compatible materials released from textiles for in vitro studies

Textile-derived fibers and particles are increasingly recognized as an important source of human and environmental exposure. While synthetic textile particles have been intensively studied as a major source of micro- and nanoplastic (MNP) pollution, textile emissions in real-world environments are far more diverse and include synthetic, natural and biobased materials. Regardless of their origin, these materials release highly complex particulate mixtures that vary widely in size, shape and chemical composition, and exhibit diverse fibrous and non-fibrous morphologies. Despite growing concern regarding the potential health impacts of released particles, current in vitro studies often lack environmental relevance as they rely on simplified, size-standardized, or pristine test materials that fail to mimic the complexity of real-world textile emissions. To address this challenge, we established a standardized, water-based Gyrowash extraction workflow to obtain sterile and environmentally relevant particle suspensions directly derived from textiles. The workflow preserved fiber integrity while providing sterile e.g. cell-culture-compatible samples that can be accurately quantified. The protocol was further optimized to enable a reproducible assessment of material-driven effects in the Calu-3 lung epithelial cell model. Optional steps for advanced physiochemical characterization are included to facilitate comprehensive material analysis and identification of potential contamination sources. Overall, this workflow provides a robust and standardized platform for obtaining representative textile-derived particle samples and supports more realistic and comparable investigations of their health effects.