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Histological Method to Study the Effect of Shear Stress on Cell Proliferation and Tissue Morphology in a Bioreactor

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Tissue Engineering and Regenerative Medicine Aims and scope

Abstract

Background:

Tissue engineering represents a promising approach for the production of bone substitutes. The use of perfusion bioreactors for the culture of bone-forming cells on a three-dimensional porous scaffold resolves mass transport limitations and provides mechanical stimuli. Despite the recent and important development of bioreactors for tissue engineering, the underlying mechanisms leading to the production of bone substitutes remain poorly understood.

Methods:

In order to study cell proliferation in a perfusion bioreactor, we propose a simplified experimental set-up using an impermeable scaffold model made of 2 mm diameter glass beads on which mechanosensitive cells, NIH-3T3 fibroblasts are cultured for up to 3 weeks under 10 mL/min culture medium flow. A methodology combining histological procedure, image analysis and analytical calculations allows the description and quantification of cell proliferation and tissue production in relation to the mean wall shear stress within the bioreactor.

Results:

Results show a massive expansion of the cell phase after 3 weeks in bioreactor compared to static control. A scenario of cell proliferation within the three-dimensional bioreactor porosity over the 3 weeks of culture is proposed pointing out the essential role of the contact points between adjacent beads. Calculations indicate that the mean wall shear stress experienced by the cells changes with culture time, from about 50 mPa at the beginning of the experiment to about 100 mPa after 3 weeks.

Conclusion:

We anticipate that our results will help the development and calibration of predictive models, which rely on estimates and morphological description of cell proliferation under shear stress.

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Acknowledgements

The authors gratefully acknowledge the LabeX LaSIPS (No. ANR-10-LABX-0040-LaSIPS) managed by the French National Research Agency under the “Investissements d’avenir” programs (Nos. ANR-11-IDEX-0003-02 and ANR-10-EQPX-37 MATMECA Grant) for their financial support.

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Author notes

  1. Morgan Chabanon

    Present address: Department of Mechanical and Aerospace Engineering, University of California San Diego, 9500 Gilman Drive, La Jolla, CA, 92093, USA

  2. Morgan Chabanon and Hervé Duval have contributed equally to this work.

Authors and Affiliations

  1. MSSMat, CentraleSupélec, Université Paris Saclay, CNRS, 3 rue Joliot-Curie, 91190, Gif-sur-Yvette, France

    Morgan Chabanon, Jérôme Grenier, Claire Beauchesne & Bertrand David

  2. LGPM, CentraleSupélec, Université Paris Saclay, 3 rue Joliot-Curie, 91190, Gif-sur-Yvette, France

    Hervé Duval & Jérôme Grenier

  3. EM2C, CentraleSupélec, Université Paris Saclay, CNRS, 3 rue Joliot-Curie, 91190, Gif-sur-Yvette, France

    Claire Beauchesne & Benoit Goyeau

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  1. Morgan Chabanon
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Correspondence to Bertrand David.

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Chabanon, M., Duval, H., Grenier, J. et al. Histological Method to Study the Effect of Shear Stress on Cell Proliferation and Tissue Morphology in a Bioreactor. Tissue Eng Regen Med 16, 225–235 (2019). https://doi.org/10.1007/s13770-019-00181-3

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