Abstract
Low intensity pulsed ultrasound (LIPUS) has been used to accelerate tissue regeneration; however, the biological mechanisms of LIPUS induced regeneration is not completely understood. The aim for this study is to elucidate the mechanical effect generated by US for the stimulation of mesenchymal stem cells (MSCs). MSCs were cultured on flexible cell culture membranes and stimulated by US for 10 min daily with acoustic intensities of 0, 6, 13.5, and 22.5 W/cm2. Cell proliferation and viability were evaluated by direct cell count and Alamar Blue assay. Morphological evaluation was performed and cell-matrix interactions were evaluated. Cell-matrix interaction was analyzed by immunochemical staining of focal adhesion proteins. LIPUS enhanced cell proliferation at higher intensities and there was an increase in cell viability after 4 consecutive days of US treatment. No morphological changes were observed in all treatments. Expression of focal adhesion protein, vinculin, was enhanced after 3 consecutive days of ultrasound treatment. Studies of media agitation did not show any enhancement effect in cell proliferation or focal adhesion protein expression. The results validates that US is able to influence the cell matrix interaction. Application of higher acoustic pressure on cell growth environment can stimulate MSC proliferation and focal adhesion.
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