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Quantifying nanotherapeutic penetration using hydrogel based microsystem as a new 3D in vitro platform

Authors :
Olivier Tillement
Ting-Di Wu
Jean-Luc Guerquin-Kern
Sandrine Lacombe
Saba Goodarzi
Charlotte Rivière
Audrey Prunet
Hélène Delanoë-Ayari
Erika Porcel
Guillaume Bort
Fabien Rossetti
François Lux
Biophysique (BIOPHYSIQUE)
Institut Lumière Matière [Villeurbanne] (ILM)
Université Claude Bernard Lyon 1 (UCBL)
Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS)-Université Claude Bernard Lyon 1 (UCBL)
Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS)
Formation, élaboration de nanomatériaux et cristaux (FENNEC)
Institut des Sciences Moléculaires d'Orsay (ISMO)
Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)
Institut Curie [Paris]
Centre National de la Recherche Scientifique (CNRS)-Université Claude Bernard Lyon 1 (UCBL)
Université de Lyon-Université de Lyon
Source :
Lab on a Chip, Lab on a Chip, Royal Society of Chemistry, 2021, 21 (13), pp.2495-2510. ⟨10.1039/D1LC00192B⟩
Publication Year :
2021
Publisher :
HAL CCSD, 2021.

Abstract

International audience; Abstract The huge gap between 2D in vitro assays used for drug screening, and the in vivo 3D-physiological environment hampered reliable predictions for the route and accumulation of nanotherapeutics in vivo. For such nanotherapeutics, Multi-Cellular Tumour Spheroids (MCTS) is emerging as a good alternative in vitro model. However, the classical approaches to produce MCTS suffer from low yield, slow process, difficulties in MCTS manipulation and compatibility with high-magnification fluorescent optical microscopy. On the other hand, spheroid-on-chip set-ups developed so far require a microfluidic practical knowledge difficult to transfer to a cell biology laboratory. We present here a simple yet highly flexible 3D-model microsystem consisting of agarose-based microwells. Fully compatible with the multi-well plates format conventionally used in cell biology, our simple process enables the formation of hundreds of reproducible spheroids in a single pipetting. Immunostaining and fluorescent imaging including live high-resolution optical microscopy can be performed in-situ , with no manipulation of spheroids. As a proof-of-principle of the relevance of such in vitro platform for nanotherapeutics evaluation, this study investigates the kinetic and localization of nanoparticles within colorectal cancer MCTS cells (HCT-116). The nanoparticles chosen are sub-5 nm ultrasmall nanoparticles made of polysiloxane and gadolinium chelates that can be visualized in MRI (AGuIX ® , currently implicated in clinical trials as effective radiosensitizers for radiotherapy) and confocal microscopy after addition of Cy 5.5. We show that the amount of AGuIX ® nanoparticles within cells is largely different in 2D and 3D. Using our flexible agarose-based microsystems, we are able to resolve spatially and temporally the penetration and distribution of AGuIX ® nanoparticles within MCTS. The nanoparticles are first found in both extracellular and intracellular space of MCTS. While the extracellular part is washed away after few days, we evidenced intracellular localisation of AGuIX ® , mainly within lysosomes compartment, but also occasionally within mitochondria. Our agarose-based microsystem appears hence as a promising 3D in vitro user-friendly platform for investigation of nanotherapeutics transport, ahead of in vivo studies. Abstract Figure Graphical abstract

Details

Language :
English
ISSN :
14730197 and 14730189
Database :
OpenAIRE
Journal :
Lab on a Chip, Lab on a Chip, Royal Society of Chemistry, 2021, 21 (13), pp.2495-2510. ⟨10.1039/D1LC00192B⟩
Accession number :
edsair.doi.dedup.....225c9a9e2ab152383aa6d1224dd7fd52