Traditional multiwell plates and petri dishes limit the evaluation of the effects of ultrasound on cells in vitro

Gupta, D., Savva, J., Li, X. , Chandler, J. H., Shelton, R. M., Scheven, B. A., Mulvana, H., Valdastri, P., Lucas, M. and Walmsley, A. D. (2022) Traditional multiwell plates and petri dishes limit the evaluation of the effects of ultrasound on cells in vitro. Ultrasound in Medicine and Biology, 48(9), pp. 1745-1761. (doi: 10.1016/j.ultrasmedbio.2022.05.001) (PMID:35760602)

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Abstract

Ultrasound accelerates healing in fractured bone; however, the mechanisms responsible are poorly understood. Experimental setups and ultrasound exposures vary or are not adequately characterized across studies, resulting in inter-study variation and difficulty in concluding biological effects. This study investigated experimental variability introduced through the cell culture platform used. Continuous wave ultrasound (45 kHz; 10, 25 or 75 mW/cm2, 5 min/d) was applied, using a Duoson device, to Saos-2 cells seeded in multiwell plates or Petri dishes. Pressure field and vibration quantification and finite-element modelling suggested formation of complex interference patterns, resulting in localized displacement and velocity gradients, more pronounced in multiwell plates. Cell experiments revealed lower metabolic activities in both culture platforms at higher ultrasound intensities and absence of mineralization in certain regions of multiwell plates but not in Petri dishes. Thus, the same transducer produced variable results in different cell culture platforms. Analysis on Petri dishes further revealed that higher intensities reduced vinculin expression and distorted cell morphology, while causing mitochondrial and endoplasmic reticulum damage and accumulation of cells in sub-G1 phase, leading to cell death. More defined experimental setups and reproducible ultrasound exposure systems are required to study the real effect of ultrasound on cells for development of effective ultrasound-based therapies not just limited to bone repair and regeneration.

Item Type:Articles
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Savva, Jill and Li, Dr Xuan and Lucas, Professor Margaret
Authors: Gupta, D., Savva, J., Li, X., Chandler, J. H., Shelton, R. M., Scheven, B. A., Mulvana, H., Valdastri, P., Lucas, M., and Walmsley, A. D.
College/School:College of Science and Engineering > School of Engineering > Systems Power and Energy
Journal Name:Ultrasound in Medicine and Biology
Publisher:Elsevier
ISSN:0301-5629
ISSN (Online):1879-291X
Published Online:24 June 2022
Copyright Holders:Copyright © 2022 The Authors
First Published:First published in Ultrasound in Medicine and Biology 48(9): 1745-1761
Publisher Policy:Reproduced under a Creative Commons License

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Project CodeAward NoProject NamePrincipal InvestigatorFunder's NameFunder RefLead Dept
300671Surgery enabled by ultrasonicsMargaret LucasEngineering and Physical Sciences Research Council (EPSRC)EP/R045291/1ENG - Systems Power & Energy