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The Journal of Membrane Biology

, Volume 250, Issue 1, pp 41–52 | Cite as

Shock Wave-Induced Damage and Poration in Eukaryotic Cell Membranes

  • Luz M. López-Marín
  • Blanca E. Millán-Chiu
  • Karen Castaño-González
  • Carmen Aceves
  • Francisco Fernández
  • Alfredo Varela-Echavarría
  • Achim M. Loske
Article

Abstract

Shock waves are known to permeabilize eukaryotic cell membranes, which may be a powerful tool for a variety of drug delivery applications. However, the mechanisms involved in shock wave-mediated membrane permeabilization are still poorly understood. In this study, the effects on both the permeability and the ultrastructural features of two human cell lineages were investigated after the application of underwater shock waves in vitro. Scanning Electron Microscopy of cells derived from a human embryo kidney (HEK)-293 and Michigan Cancer Foundation (MCF)-7 cells, an immortalized culture derived from human breast adenocarcinoma, showed a small amount of microvilli (as compared to control cells), the presence of hole-like structures, and a decrease in cell size after shock wave exposure. Interestingly, these effects were accompanied by the permeabilization of acid and macromolecular dyes and gene transfection. Trypan blue exclusion assays indicated that cell membranes were porated during shock wave treatment but resealed after a few seconds. Deformations of the cell membrane lasted for at least 5 min, allowing their observation in fixed cells. For each cell line, different shock wave parameters were needed to achieve cell membrane poration. This difference was correlated to successful gene transfection by shock waves. Our results demonstrate, for the first time, that shock waves induce transient micro- and submicrosized deformations at the cell membrane, leading to cell transfection and cell survival. They also indicate that ultrastructural analyses of cell surfaces may constitute a useful way to match the use of shock waves to different cells and settings.

Keywords

Cell transfection Shock waves Acoustic cavitation 

Abbreviations

DMEM

Dulbecco’s modified Eagle’s medium

FACS

Fluorescence-assisted cell sorting

FBS

Fetal bovine serum

FITC

Fluorescein isothiocyanate

FSC

Forward scatter

GFP

Green fluorescent protein

HEK

Human embryo kidney

MCF

Michigan Cancer Foundation

PBS

Phosphate-buffered saline

SEM

Scanning Electron Microscopy

Notes

Acknowledgments

The authors would like to thank Lourdes Palma-Tirado, Alicia del Real, and Carmen Peza for support with electron microscopy; Nydia Hernández for assistance with confocal microscopy; Carlos Castellanos-Barba for help with FACS; and Paula Bernardino, Anaid Antaramian, Adriana González-Gallardo, and Guillermo Vázquez for excellent technical assistance. This research was supported by the National University of Mexico (UNAM), through grant PAPIIT-DGAPA IT200615 and by the Science and Technology Council—Government of Querétaro, Mexico (CONCYTEQ), through its “Nuevos Talentos en Ciencia y Tecnología” program. B.E.M-C. was a fellow from the DGAPA postdoctoral program (UNAM) during the initial stage of this study.

Supplementary material

232_2016_9921_MOESM1_ESM.docx (164 kb)
Supplementary material 1 (DOCX 164 kb) Fig. S1 HEK-293 cell proliferation after exposure to shock waves. Cells (1 × 106) were treated with 0, 60, 120 or 180 pulses. Then, 2 × 105 cells were seeded in 6-cm-diameter dishes and cultured for 24 or 48 hs in DMEM supplemented with 10 % FBS. After incubation, the medium was discarded, cells were washed with 1.0 ml PBS, detached by treatment with 0.3 ml Trypsin–EDTA solution for 5 min. After blocking trypsin with 0.7 ml DMEM with 10 % FBS, the suspended cells were counted in a Neubauer chamber. The experiments were performed in triplicates. No statistical difference was found between groups of cells treated with different doses of shock waves, incubated during equivalent periods. Fig. S2 Green fluorescent protein-expressing MCF-7 cells after shock wave-mediated transfection. Shock waves were applied to MCF-7 suspended cells in the presence of the pCX::GFPGPI2 plasmid. The images show typical overviews of cells treated with 0 (a) and 125 shock waves (b), incubated onto Fluorodish plates, and visualized by epifluorescence microscopy.

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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Luz M. López-Marín
    • 1
  • Blanca E. Millán-Chiu
    • 2
  • Karen Castaño-González
    • 1
  • Carmen Aceves
    • 3
  • Francisco Fernández
    • 1
  • Alfredo Varela-Echavarría
    • 3
  • Achim M. Loske
    • 1
  1. 1.Centro de Física Aplicada y Tecnología AvanzadaUniversidad Nacional Autónoma de MéxicoQuerétaroMexico
  2. 2.CONACYT - Centro de Física Aplicada y Tecnología AvanzadaUniversidad Nacional Autónoma de MéxicoQuerétaroMexico
  3. 3.Instituto de NeurobiologíaUniversidad Nacional Autónoma de MéxicoQuerétaroMexico

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