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Channeling Effect and Tissue Morphology in a Perfusion Bioreactor Imaged by X-Ray Microtomography

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Tissue Engineering and Regenerative Medicine Aims and scope

Abstract

Background:

Perfusion bioreactors for tissue engineering hold great promises. Indeed, the perfusion of culture medium enhances species transport and mechanically stimulates the cells, thereby increasing cell proliferation and tissue formation. Nonetheless, their development is still hampered by a lack of understanding of the relationship between mechanical cues and tissue growth.

Methods:

Combining tissue engineering, three-dimensional visualization and numerical simulations, we analyze the morphological evolution of neo-tissue in a model bioreactor with respect to the local flow pattern. NIH-3T3 cells were grown under perfusion for one, two and three weeks on a stack of 2 mm polyacetal beads. The model bioreactor was then imaged by X-ray micro-tomography and local tissue morphology was analyzed. To relate experimental observations and mechanical stimulii, a computational fluid dynamics model of flow around spheres in a canal was developed and solved using the finite element method.

Results:

We observe a preferential tissue formation at the bioreactor periphery, and relate it to a channeling effect leading to regions of higher flow intensity. Additionally, we find that circular crater-like tissue patterns form in narrow channel regions at early culture times. Using computational fluid dynamic simulations, we show that the location and morphology of these patterns match those of shear stress maxima. Finally, the morphology of the tissue is qualitatively described as the tissue grows and reorganizes itself.

Conclusion:

Altogether, our study points out the key role of local flow conditions on the tissue morphology developed on a stack of beads in perfusion bioreactors and provides new insights for effective design of hydrodynamic bioreactors for tissue engineering using bead packings.

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Acknowledgements

This work has been financially supported by the French Agence Nationale de la Recherche through the Investissements d'avenir program (ANR-10-EQPX-37 MATMECA Grant) and the Fondation pour la Recherche Médicale (FRM FDT201805005480). We thank Pascal Silberzan (Institut Curie) for kindly providing the NIH-3T3 cell line and Stéphane Roux (LMT, ENS Paris-Saclay) for his kind help regarding image post-processing.

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Author notes

  1. Benoît Goyeau and Bertrand David should be considered joint senior authors.

Authors and Affiliations

  1. Lab. EM2C, UPR CNRS 288, CentraleSupélec, Université Paris-Saclay, 3 rue Joliot-Curie, 91192, Gif-sur-Yvette Cedex, France

    Claire C. Beauchesne & Benoît Goyeau

  2. Lab. MSSMat, UMR CNRS 8579, CentraleSupélec, Université Paris-Saclay, 3 rue Joliot-Curie, 91192, Gif-sur-Yvette Cedex, France

    Claire C. Beauchesne & Bertrand David

  3. Single Molecule Biophotonics Lab. ICFO, The Institute of Photonic Sciences, av. Carl Friedrich Gauss, 3, 08860, Castelldefels, Barcelona, Spain

    Morgan Chabanon

  4. ENS Paris Saclay, LMT, CNRS, UMR 8535, 61 avenue du Président Wilson, 94230, Cachan, France

    Benjamin Smaniotto

  5. Institut Jacques Monod (IJM), UMR CNRS 7592, Université Paris Diderot, 15 rue Hélène Brion, 75013, Paris, France

    Benoît Ladoux

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  1. Claire C. Beauchesne
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Correspondence to Benoît Goyeau.

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Beauchesne, C.C., Chabanon, M., Smaniotto, B. et al. Channeling Effect and Tissue Morphology in a Perfusion Bioreactor Imaged by X-Ray Microtomography. Tissue Eng Regen Med 17, 301–311 (2020). https://doi.org/10.1007/s13770-020-00246-8

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