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Design of a cone-and-plate device for controlled realistic shear stress stimulation on endothelial cell monolayers

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Abstract

Endothelial cells are constantly exposed to blood flow and the resulting frictional force, the wall shear stress, varies in magnitude and direction with time, depending on vasculature geometry. Previous studies have shown that the structure and function of endothelial cells, and ultimately of the vessel wall, are deeply affected by the nature of wall shear stress waveforms. To investigate the in vitro effects of these stimuli, we developed a compact, programmable, real-time operated system based on cone-and-plate geometry, that can be used within a standard cell incubator. To verify the capability to replicate realistic shear stress waveforms, we calculated both analytically and numerically to what extent the system is able to correctly deliver the stimuli defined by the user at plate level. Our results indicate that for radii greater than 25 mm, the shear stress is almost uniform and directly proportional to cone rotation velocity. We further established that using a threshold of 10 Hz of wall shear stress waveform frequency components, oscillating flow conditions can be reproduced on cell monolayer surface. Finally, we verified the capability of the system to perform long-term flow exposure experiments ensuring sterility and cell culture viability on human umbilical vein endothelial cells exposed to unidirectional and oscillating shear stress. In conclusion, the system we developed is a highly dynamic, easy to handle, and able to generate pulsatile and unsteady oscillating wall shear stress waveforms. This system can be used to investigate the effects of realistic stimulations on endothelial cells, similar to those exerted in vivo by blood flow.

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Acknowledgments

This research was supported by the Cariplo Foundation (Fondazione Cariplo, Milano, Italy) under the “I4BIO—Innovazione e Bioingegneria” Project. Marco Franzoni was the recipient of a research fellowship from the ARMR Foundation (Aiuti per la Ricerca sulle Malattie Rare, Bergamo, Italy).

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Authors and Affiliations

  1. Department of Biomedical Engineering, IRCCS - Istituto di Ricerche Farmacologiche “Mario Negri”, Via Stezzano, 87, 24126, Bergamo, Italy

    Marco Franzoni, Irene Cattaneo, Bogdan Ene-Iordache & Andrea Remuzzi

  2. Department of Management, Information and Production Engineering, University of Bergamo, Viale Marconi 4, 24144, Dalmine, BG, Italy

    Alberto Oldani, Paolo Righettini & Andrea Remuzzi

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  1. Marco Franzoni
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  2. Irene Cattaneo
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  3. Bogdan Ene-Iordache
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  4. Alberto Oldani
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  5. Paolo Righettini
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  6. Andrea Remuzzi
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Correspondence to Andrea Remuzzi.

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All the authors have declared no competing interests.

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Franzoni, M., Cattaneo, I., Ene-Iordache, B. et al. Design of a cone-and-plate device for controlled realistic shear stress stimulation on endothelial cell monolayers. Cytotechnology 68, 1885–1896 (2016). https://doi.org/10.1007/s10616-015-9941-2

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  • DOI: https://doi.org/10.1007/s10616-015-9941-2

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