Structural and permeability sensitivity of cells to low intensity ultrasound: Infrared and fluorescence evidence in vitro |
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| Institution: | 1. Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139, USA;2. The David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA 02139, USA;3. Department of Biology, Massachusetts Institute of Technology, Cambridge, MA 02139, USA;2. Department of Biology, Concordia University, Montreal, Quebec, Canada;3. Center for Ultrasound Molecular Imaging and Therapeutics, University of Pittsburgh, Pittsburgh, Pennsylvania, USA;4. Department of Cell Biology, Center for Biologic Imaging, University of Pittsburgh, Pittsburgh, Pennsylvania, USA;5. Pittsburgh Heart and Vascular Medicine Institute, University of Pittsburgh, Pittsburgh, Pennsylvania, USA |
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| Abstract: | This work is focused on the in vitro study of the effects induced by medical ultrasound (US) in murine fibroblast cells (NIH-3T3) at a low-intensity of exposure (spatial peak temporal average intensity Ita < 0.1 W cm?2). Conventional 1 MHz and 3 MHz US devices of therapeutic relevance were employed with varying intensity and exposure time parameters. In this framework, upon cells exposure to US, structural changes at the molecular level were evaluated by infrared spectroscopy; alterations in plasma membrane permeability were monitored in terms of uptake efficiency of small cell-impermeable model drug molecules, as measured by fluorescence microscopy and flow cytometry. The results were related to the cell viability and combined with the statistical PCA analysis, confirming that NIH-3T3 cells are sensitive to therapeutic US, mainly at 1 MHz, with time-dependent increases in both efficiency of uptake, recovery of wild-type membrane permeability, and the size of molecules entering 3T3. On the contrary, the exposures from US equipment at 3 MHz show uptakes comparable with untreated samples. |
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| Keywords: | Ultrasound Membrane sonoporation Cell uptake FTIR spectroscopy Fluorescence microscopy |
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