Multimodal characterization of the safety and sustainability of cellulose nanofibers in 2D and 3D in vitro models [* Cross-Reference *]

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Antje Biesemeier; Pablo Delfino; Iris Riemann; Nicole Jost; Y. Kohl, 2026, "Multimodal characterization of the safety and sustainability of cellulose nanofibers in 2D and 3D in vitro models [* Cross-Reference *]", https://lore.list.lu/citation?persistentId=perma:LIST.XGTLAY, LIST Open Repository, V1
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Description
As part of SSbD4CheM project (PI Milica Velimirovic, VITO), this study focuses on the development of focused ion beam secondary ion mass spectrometry (FIB-SIMS) for the investigation of 2D and 3D in vitro models after exposure to cellulose nanofibers (CNC). In the past, chemical imaging facilitated the correlative investigation of ultrastructural changes after exposure to e.g. metallic nanoparticles and at the same time, informed about the identity and localization of the ingested materials based on epoxy cross sections [1]. Here, a human 2D (HT29 on a porous cell culture insert) and respective 3D-colorectal adenocarcinoma cell model (HT29-MTX-E12 spheroids) were exposed to CNC submerged for 24h (Celluloselab CNC-Cationic 1.00% H2O 98.80 %, 1,2-hexanediol 0.10%, 4'hydroxyacetophenone 0.10%, if not indicated different).Biological endpoints like cell viability, membrane leakage and cell vitality were determined and samples were prepared for focused ion beam-based secondary ion mass spectrometry (FIB-SIMS) for ultrastructural and chemical characterization. The LIST magnetic sector SIMS system equipped with a continuous focal plane detector technology allows parallel detection of all masses for each single pixel in the scanned region and directly links SIMS elemental mappings with other signals obtained from the same FIB microscopy platform (e.g. secondary, transmitted or back-scattered electrons and/or ions [2,3]) with unprecedented spatial resolution and sensitivity. Cationic CNC exposure was not toxic in the investigated concentrations. For more in depth physico-chemical and ultrastructural analysis, we present two different preparation steps. Epoxy-embedding and cross-sectioning yields flat samples suitable for SIMS analysis. However, introduction of additional organic information coming from the resin interferes with signals from cellular and CNC components. On the other hand, resin-free critical point drying allows near-to native chemical identification of the samples. However, high topography leads to signal shadowing, edge effects and potentially decreased conductivity after sputter-coating. Future investigations will include micro-machining of highly topographic materials for resin-free flat sample preparations. Multi-peak fingerprinting of different CNC classes will allow clear identification of CNC signals within exposed matrices. (2026-04-01)
***This entry has been automatically imported via OpenAlex by LIST harvest scripts. Please refer to https://doi.org/10.5281/zenodo.19368540 for the original and latest version of the publication*** (2026-07-01)
Subject Engineering; Physics
Keyword Secondary ion mass spectrometry, Focused ion beam, Nanofiber, Cellulose, Scanning electron microscope, Characterization (materials science), Mass spectrometry, Ion beam, Nanoparticle
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