Traditional multiwell plates and petri dishes limit the evaluation of the effects of ultrasound on cells in vitro
Gupta, Dhanak and Savva, Jill and Li, Xuan and Chandler, James H and Shelton, Richard M and Scheven, Ben A and Mulvana, Helen and Valdastri, Pietro and Lucas, Margaret and Walmsley, A Damien (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. ISSN 0301-5629 (https://doi.org/10.1016/j.ultrasmedbio.2022.05.001)
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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/cm 2, 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.
ORCID iDs
Gupta, Dhanak, Savva, Jill, Li, Xuan, Chandler, James H, Shelton, Richard M, Scheven, Ben A, Mulvana, Helen ORCID: https://orcid.org/0000-0002-5058-0299, Valdastri, Pietro, Lucas, Margaret and Walmsley, A Damien;-
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Item type: Article ID code: 80624 Dates: DateEvent30 September 2022Published24 June 2022Published Online2 May 2022AcceptedSubjects: Technology > Engineering (General). Civil engineering (General) > Bioengineering Department: Faculty of Engineering > Biomedical Engineering Depositing user: Pure Administrator Date deposited: 10 May 2022 10:20 Last modified: 05 Sep 2024 01:13 URI: https://strathprints.strath.ac.uk/id/eprint/80624