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Analysis of Fluid Flow and Wall Shear Stress Patterns Inside Partially Filled Agitated Culture Well Plates

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Abstract

The appearance of highly resistant bacterial biofilms in both community and hospitals environments is a major challenge in modern clinical medicine. The biofilm structural morphology, believed to be an important factor affecting the behavioral properties of these “super bugs”, is strongly influenced by the local hydrodynamics over the microcolonies. Despite the common use of agitated well plates in the biology community, they have been used rather blindly without knowing the flow characteristics and influence of the rotational speed and fluid volume in these containers. The main purpose of this study is to characterize the flow in these high-throughput devices to link local hydrodynamics to observed behavior in cell cultures. In this work, the flow and wall shear stress distribution in six-well culture plates under planar orbital translation is simulated using Computational Fluid Dynamics (CFD). Free surface, flow pattern and wall shear stress for two shaker speeds (100 and 200 rpm) and two volumes of fluid (2 and 4 mL) were investigated. Measurements with a non-intrusive optical shear stress sensor and High Frame-rate Particle Imaging Velocimetry (HFPIV) are used to validate CFD predictions. An analytical model to predict the free surface shape is proposed. Results show a complex three-dimensional flow pattern, varying in both time and space. The distribution of wall shear stress in these culture plates has been related to the topology of flow. This understanding helps explain observed endothelial cell orientation and bacterial biofilm distributions observed in culture dishes. The results suggest that the mean surface stress field is insufficient to capture the underlying dynamics mitigating biological processes.

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Acknowledgments

The authors recognize valuable discussions with Dr. Victoria Kostenko. M. Mehdi Salek would like to acknowledge Alberta Ingenuity Fund (AIF), now part of Alberta Innovates Technology Futures for their support through a scholarship. This research has been enabled by the use of computing resources provided by WestGrid and Compute/Calcul Canada.

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Authors and Affiliations

  1. Biofilm Engineering Research Group, Calgary Center for Innovative Technology, University of Calgary, Calgary, AB, Canada

    M. Mehdi Salek & Robert J. Martinuzzi

  2. Department of Mechanical and Manufacturing Engineering, University of Calgary, 2500 University Drive N.W., Calgary, AB, T2N 1N4, Canada

    M. Mehdi Salek, Pooria Sattari & Robert J. Martinuzzi

Authors
  1. M. Mehdi Salek
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  2. Pooria Sattari
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  3. Robert J. Martinuzzi
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Correspondence to Robert J. Martinuzzi.

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Associate Editor Konstantinos Konstantopoulo oversaw the review of this article.

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Salek, M.M., Sattari, P. & Martinuzzi, R.J. Analysis of Fluid Flow and Wall Shear Stress Patterns Inside Partially Filled Agitated Culture Well Plates. Ann Biomed Eng 40, 707–728 (2012). https://doi.org/10.1007/s10439-011-0444-9

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  • DOI: https://doi.org/10.1007/s10439-011-0444-9

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