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bioRxiv · 10.1101/2024.11.15.623765

Tumor Spheroid Uptake of Fluorescent Nanodiamonds is Limited by Mass Density: a 4D Light-Sheet Assay

Niora, M.·Dufva, M.·Jauffred, L.·Berg-Sorensen, K.
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Abstract

Fluorescent nanodiamonds (FNDs) with nitrogen-vacancy centers are promising candidates for long-term biolabeling and biosensing applications due to their biocompatibility and unique optomagnetic properties. The employment of nanomaterials in cancer therapy and diagnostics, requires a deep understanding of how nanoparticles (NPs) interact with the three-dimensional (3D) tumor environment. We developed a novel approach, the "Tumor-in-a-Tube" platform, using 4D light-sheet microscopy to explore the spatio-temporal dynamics of FNDs with 3D tumor spheroids. By monitoring the real-time NP sedimentation, spheroid penetration, and cellular uptake of FNDs and polystyrene nanoparticles (PNPs), we marked the impact of NP mass density on their spheroid interaction. Unlike PNPs, higher-density FNDs underwent rapid sedimentation, which minimized their effective concentration and hindered the FND - spheroid interactions. This results in constrained intratumoral accumulation, and size-independent uptake and penetration. Longer FND effective-exposure-time promotes size-dependent cell uptake, verified by FND treatment on 2D monolayers. Nonetheless, FNDs exhibited good biocompatibility and long-term spheroid labeling, allowing for cell isolation from different spheroid layers. Our results suggest the need for NP effective-exposure-time calibration in comparative NP assays, in 3D static models. Overall, our platform provides a valuable tool for bridging the gap between 2D and 3D static models in NP assessment, drug delivery, toxicology profiling and translational research.

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Niora, M., Dufva, M., Jauffred, L., Berg-Sorensen, K.. 2024-11-15. Tumor Spheroid Uptake of Fluorescent Nanodiamonds is Limited by Mass Density: a 4D Light-Sheet Assay. https://doi.org/10.1101/2024.11.15.623765

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