Optimal Ultrasound Exposure Conditions for Maximizing C2C12 Muscle Cell Proliferation and Differentiation.

Author: Salgarella AR1, Cafarelli A2, Ricotti L2, Capineri L3, Dario P2, Menciassi A2
Affiliation: <sup>1</sup>The BioRobotics Institute, Scuola Superiore Sant'Anna, Pontedera (Pisa), Italy. Electronic address: a.salgarella@santannapisa.it. <sup>2</sup>The BioRobotics Institute, Scuola Superiore Sant'Anna, Pontedera (Pisa), Italy. <sup>3</sup>Department of Information Engineering, University of Florence, Florence, Italy.
Conference/Journal: Ultrasound Med Biol.
Date published: 2017 Apr 19
Other: Pages: S0301-5629(17)30103-5 , Special Notes: doi: 10.1016/j.ultrasmedbio.2017.03.003. [Epub ahead of print] , Word Count: 179


Described here is an in vitro systematic investigation of the effects on C2C12 myoblasts of exposure to finely controlled and repeatable low-intensity pulsed ultrasound of different frequencies (500 kHz, 1 MHz, 3 MHz and 5 MHz) and different intensities (250, 500 and 1000 mW/cm2). An in-house stimulation system and an ultrasound-transparent cell culture well minimized reflections and attenuations, allowing precise control of ultrasound delivery. Results indicated that a 3 MHz stimulation at 1 W/cm2 intensity maximized cell proliferation in comparison with the other exposure conditions and untreated controls. In contrast, cell differentiation and the consequent formation of multinucleated myotubes were maximized by 1 MHz stimulation at 500 mW/cm2 intensity. The highly controlled exposure conditions employed allowed precise correlation of the ultrasound delivery to the bio-effects produced, thus overcoming the inconsistency of some results available in the literature and contributing to the potential of ultrasound treatment for muscle therapy and regeneration.

Copyright © 2017 World Federation for Ultrasound in Medicine & Biology. Published by Elsevier Inc. All rights reserved.

KEYWORDS: Bio-effects; C2C12; Differentiation; Frequency; Intensity; Low-intensity pulsed ultrasound; Muscle regeneration; Myoblasts; Proliferation; Ultrasound stimulation

PMID: 28433437 DOI: 10.1016/j.ultrasmedbio.2017.03.003