Traditional Multiwell Plates and Petri Dishes Limit the Evaluation of the Effects of Ultrasound on Cells In Vitro

Gupta, Dhanak; Savva, Jill; Li, Xuan; Chandler, James H.; Shelton, Richard M.; Scheven, Ben A.; Mulvana, Helen; Valdastri, Pietro; Lucas, Margaret; Walmsley, A. D.

doi:10.1016/j.ultrasmedbio.2022.05.001

Cited by 6 publications

(5 citation statements)

References 38 publications

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“…Limited attempts to calculate US propagation in such culturing vessels have been carried out to date. For example, the finite-element method (FEM) was used to calculate the acoustic pressures and shear wave vibration displacements on the surfaces of six-well plates when a transducer is inserted into an individual well [32]. Although these calculations were limited to a frequency of 45 kHz, FEM was also able to demonstrate the establishment of SW patterns, which is a known occurrence in these culturing vessel types.…”

Section: Discussionmentioning

confidence: 99%

Towards an understanding of the mechanisms of therapeutic ultrasound on biomimetic models of cancer

Silva,

Cheema,

Gélat

2024

J. Phys.: Conf. Ser.

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Therapeutic ultrasound is transforming the treatment of a range of malignancies in a non-invasive and non-systemic manner. Low-intensity ultrasound (LIUS) has been proposed to selectively eradicate cancer cells but the underlying biological mechanisms remains unknown. To gain an understanding of this phenomenon, 2D breast cancer monocultures were sonicated at varying acoustic intensities (0.1–0.5 W·cm−2) and excitation times (1–10 minutes). Additionally, 2D monocultures consisting of healthy cell lines were sonicated at varying acoustic intensities (0.1–0.5 W·cm−2) to determine any distinguishing biological responses. To begin recapitulating in vivo conditions, breast cancer cells were also seeded into 3D collagen hydrogels. At a 1 MHz frequency, 20% duty cycle, 100 Hz pulse repetition frequency, a significant drop in cancer cell viability is observed at a sonication intensity of 0.5 W·cm−2 and over 10 minute excitation time. Healthy counterparts subjected to the same parameters revealed no distinguishing effects. Sonication of breast cancer cells seeded in 3D collagen hydrogels revealed no effect in cell viability compared to non-sonicated controls. The acoustic wave propagation software OptimUS was used to determine the influence culturing plates have on ultrasound propagation, revealing these materials can significantly vary the acoustic field at frequencies relevant to LIUS.

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Section: Discussionmentioning

confidence: 99%

Towards an understanding of the mechanisms of therapeutic ultrasound on biomimetic models of cancer

Silva,

Cheema,

Gélat

2024

J. Phys.: Conf. Ser.

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“…It is also worth mentioning that, in general, US stimulations (alone or in synergy with piezoelectric materials) are typically poorly controlled in the state-of-art: in most of the in vitro studies, the setups used for stimulating cells are affected by poorly standardized configurations, lack of proper calibration, and lack of control on US wave reflections/attenuations. These aspects jeopardize the reliability of many studies and slow down their possible future clinical translation . A few studies focused on electric charges generated by vibrating quartz coverslips to induce MSC chondrogenesis. , However, the exploited mechanism was quite different in this case.…”

Section: Introductionmentioning

confidence: 99%

“…These aspects jeopardize the reliability of many studies and slow down their possible future clinical translation. 22 A few studies focused on electric charges generated by vibrating quartz coverslips to induce MSC chondrogenesis. 23,24 However, the exploited mechanism was quite different in this case.…”

Section: Introductionmentioning

confidence: 99%

Ultrasound Stimulation of Piezoelectric Nanocomposite Hydrogels Boosts Chondrogenic Differentiation in Vitro, in Both a Normal and Inflammatory Milieu

Ricotti,

Cafarelli,

Manferdini

et al. 2024

ACS Nano

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The use of piezoelectric nanomaterials combined with ultrasound stimulation is emerging as a promising approach for wirelessly triggering the regeneration of different tissue types. However, it has never been explored for boosting chondrogenesis. Furthermore, the ultrasound stimulation parameters used are often not adequately controlled. In this study, we show that adipose-tissue-derived mesenchymal stromal cells embedded in a nanocomposite hydrogel containing piezoelectric barium titanate nanoparticles and graphene oxide nanoflakes and stimulated with ultrasound waves with precisely controlled parameters (1 MHz and 250 mW/cm 2 , for 5 min once every 2 days for 10 days) dramatically boost chondrogenic cell commitment in vitro. Moreover, fibrotic and catabolic factors are strongly down-modulated: proteomic analyses reveal that such stimulation influences biological processes involved in cytoskeleton and extracellular matrix organization, collagen fibril organization, and metabolic processes. The optimal stimulation regimen also has a considerable anti-inflammatory effect and keeps its ability to boost chondrogenesis in vitro, even in an inflammatory milieu. An analytical model to predict the voltage generated by piezoelectric nanoparticles invested by ultrasound waves is proposed, together with a computational tool that takes into consideration nanoparticle clustering within the cell vacuoles and predicts the electric field streamline distribution in the cell cytoplasm. The proposed nanocomposite hydrogel shows good injectability and adhesion to the cartilage tissue ex vivo, as well as excellent biocompatibility in vivo, according to ISO 10993. Future perspectives will involve preclinical testing of this paradigm for cartilage regeneration.

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“…In general, these effects have been studied using cell cultures and commercially available transducers. However, these transducers are often quite large, are not easily adaptable in terms of operating conditions, and cannot be used simultaneously in multiple culture wells, which can lead to experimental variability [19]. Therefore, ultrasound devices specifically designed for use in cell culture experiments are of great interest.…”

Section: Introductionmentioning

confidence: 99%

Exploring the Potential of Ultrasound Therapy to Reduce Skin Scars: An In Vitro Study Using a Multi-Well Device Based on Printable Piezoelectric Transducers

Riis Porsborg,

Krzyslak,

Pierchala

et al. 2023

Bioengineering

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Excessive skin scarring affects over 100 million patients worldwide, with effects ranging from cosmetic to systemic problems, and an effective treatment is yet to be found. Ultrasound-based therapies have been used to treat a variety of skin disorders, but the exact mechanisms behind the observed effects are still unclear. The aim of this work was to demonstrate the potential of ultrasound for the treatment of abnormal scarring by developing a multi-well device based on printable piezoelectric material (PiezoPaint™). First, compatibility with cell cultures was evaluated using measurements of heat shock response and cell viability. Second, the multi-well device was used to treat human fibroblasts with ultrasound and quantify their proliferation, focal adhesions, and extracellular matrix (ECM) production. Ultrasound caused a significant reduction in fibroblast growth and ECM deposition without changes in cell viability or adhesion. The data suggest that these effects were mediated by nonthermal mechanisms. Interestingly, the overall results suggest that ultrasound treatment would a be beneficial therapy for scar reduction. In addition, it is expected that this device will be a useful tool for mapping the effects of ultrasound treatment on cultured cells.

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Traditional Multiwell Plates and Petri Dishes Limit the Evaluation of the Effects of Ultrasound on Cells In Vitro

Cited by 6 publications

References 38 publications

Towards an understanding of the mechanisms of therapeutic ultrasound on biomimetic models of cancer

Towards an understanding of the mechanisms of therapeutic ultrasound on biomimetic models of cancer

Ultrasound Stimulation of Piezoelectric Nanocomposite Hydrogels Boosts Chondrogenic Differentiation in Vitro, in Both a Normal and Inflammatory Milieu

Exploring the Potential of Ultrasound Therapy to Reduce Skin Scars: An In Vitro Study Using a Multi-Well Device Based on Printable Piezoelectric Transducers

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