Low-Intensity Pulsed Ultrasound Effect on MIO-M1 Cell Viability: Setup Validation and Standing Waves Analysis

Irais Poblete-Naredo,Mario Ibrahin Gutierrez,Arnulfo Albores,Josefina Gutiérrez-Martínez,Diana Estela Mendoza-Sánchez,Lorenzo Leija,Arturo Ortega,Arturo Vera

doi:10.3390/app11010271

Irais Poblete-Naredo, Mario Ibrahin Gutierrez + Show 6 more

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https://doi.org/10.3390/app11010271

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Abstract

Low-intensity pulsed ultrasound (LIPUS) has been proposed for novel therapies still under study, where similar parameters and protocols have been used for producing opposite effects that range from increasing cell viability to provoking cell death. Those divergent outcomes make the generalization of expected effects difficult for cell models not yet studied. This paper presents the effect of LIPUS on the viability of the MIO-M1 cell line for two well-established setups and different protocols; the acoustic intensities, duty factors, and treatment duration were varied. Measurements and models for acoustic and thermal analysis are included for proposing a solution to improve the reproducibility of this kind of experiments. Results indicate that MIO-M1 viability is less affected for the cells treated through a dish that is partially immersed in water; in these conditions, the cells neither show detrimental nor proliferative effects at intensities lower than 0.4 W/cm2 at 20% duty factor. However, cell viability was reduced when LIPUS was followed by cell subculturing. Treating the cells through a gel, with the culture dish placed on the transducer, increases cell mortality by the production of standing waves and mixed vibration-acoustical effects. Using the water-based setup with a 1° dish inclination reduces the effects of standing waves.

Highlights

Introduction nal affiliationsUltrasound has been proposed for an extensive variety of therapies with different modalities and intensities for producing either thermal or non-thermal effects
In vitro experiments were performed to evaluate the acoustic effect on the viability of the cell line MIO-M1
Cell death was significant only at 1.6 W/cm2 at 20% duty factors (DF) (+ p < 0.05, post-hoc Dunn’s test) or 50% DF (** p < 0.01, post-hoc Dunnett’s test), with reductions on cell viability to 64.4% and 14.7%, respectively

Summary

Introduction

Introduction nal affiliationsUltrasound has been proposed for an extensive variety of therapies with different modalities and intensities for producing either thermal or non-thermal effects. The lowintensity pulsed ultrasound (LIPUS) is a type of non-thermal ultrasound modality with intensities lower than 3 W/cm applied in a pulsed regime that has been proposed for multiple medical applications [1,2,3,4,5,6,7]. Those therapeutic applications range from tissue healing and cell survival stimulation to cell death induction (cancer therapy), in which comparable acoustic treatment conditions are used for producing opposite effects.

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