Skip to main content
SpringerLink
Log in

Use of EPR and FTIR to detect biological effects of ultrasound and microbubbles on a fibroblast cell line

  • Original Paper
  • Published:
European Biophysics Journal Aims and scope Submit manuscript

Abstract

Structural and functional effects of exposing murine fibroblasts (NIH 3T3) to therapeutic ultrasound at 1 MHz frequency are described. These bioeffects can be attributed to the formation of free radical species by sonolysis of water. When cavitation occurs, dissociation of water vapor into H atoms and OH radicals is observed; these H atoms and OH radicals combine to form H2, H2O2, and HO2. The radicals can chemically modify biomolecules, for example enzymes, DNA, and lipids. Generation of free radicals during exposure to ultrasound with or without encapsulated microbubbles (contrast agents) was studied by use of electron paramagnetic resonance with DMPO spin trapping. Recently the potential for possible use of these microbubbles in gene therapy has been investigated, because of the ability of the stabilized microbubbles to release their content when exposed to ultrasound. Structural changes were studied by Fourier-transform infrared spectroscopy, and induction of possible genotoxic damage by exposure of the cells to therapeutic ultrasound at 1 MHz frequency with our experimental device was verified by use of the cytokinesis-block micronucleus assay.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  • Barnett S (1998) Free radical production: its biological consequences. Ultrasound Med Biol 24(Suppl 1):S29–S34

    Google Scholar 

  • Bekeredjian R, Chen S, Frenkel PA, Grayburn PA, Shohet RV (2003) Ultrasound-targeted microbubble destruction can repeatedly direct highly specific plasmid expression to the heart. Circulation 108:1022–1026

    Article  PubMed  Google Scholar 

  • Christiansen JP, French BA, Klibanov AL, Kaul S, Lindner JR (2003) Targeted tissue transfection with ultrasound destruction of plasmid-bearing cationic microbubbles. Ultrasound Med Biol 29:1759–1767

    Article  PubMed  Google Scholar 

  • Conti L, Grimaldi P, Udroiu I, Bedini A, Giliberti C, Giuliani L, Palomba R, Congiu Castellano A (2010) Effects induced in cells by ultrasound revealed by ATR-FTIR spectroscopy. Vib Spectrosc 52:79–84

    Article  CAS  Google Scholar 

  • Di Giambattista L, Grimaldi P, Udroiu I, Pozzi D, Cinque G, Frogley M, Giansanti A, Congiu Castellano A (2009) FTIR spectral imaging as a probe of ultrasound effect on cells in vitro. Biophys Bioeng Lett 2:2

    Google Scholar 

  • Di Giambattista L, Grimaldi P, Gaudenzi S, Pozzi D, Grandi M, Morrone S, Silvestri I, Congiu Castellano A (2010) UVB radiation induced effects on cells studied by FTIR spectroscopy. Eur Biophys J 39:929

    Article  PubMed  CAS  Google Scholar 

  • Di Giambattista L, Pozzi D, Grimaldi P, Gaudenzi S, Morrone S, Congiu Castellano A (2011) New marker of tumor cell death revealed by ATR-FTIR spectroscopy. Anal Bioanal Chem 399(8):2771–2778. doi:10.1007/s00216-011-4654-7

    Google Scholar 

  • Draux F, Jeannesson P, Gobinet C, Sule-Suso J, Pijanka J, Sandt C, Dumas P, Manfait M, Sockalingum GD (2009) IR spectroscopy reveals effect of non-cytotoxic doses of anti-tumour drug on cancer cells. Anal Bioanal Chem 395:2293–2301

    Article  PubMed  CAS  Google Scholar 

  • Gasper R, Dewelle J, Kiss R, Mijatovic T, Goormaghtigh E (2009) IR spectroscopy as a new tool for evidencing antitumor drug signatures. Biochim Biophys Acta 1788:1263–1270

    Article  PubMed  CAS  Google Scholar 

  • Hiraoka W, Honda H, Feril LB Jr, Kudo N, Kondo T (2006) Comparison between sonodynamic effect and photodynamic effect with photosensitizers on free radical formation and cell killing. Ultrasound Sonochem 13:535–542

    Article  CAS  Google Scholar 

  • Juffermans LJM, Dijkmans PA, Musters RJP, Visser CA, Kamp O (2006) Transient permeabilization of cell membranes by ultrasound-exposed microbubbles is related to formation of hydrogen peroxide. Am J Physiol Heart Circ Physiol 291:H1595–H1601

    Article  PubMed  CAS  Google Scholar 

  • Kirsch-Volders M, Fenech M (2001) Inclusion of micronuclei in non-divided mononuclear lymphocytes and necrosis/apoptosis may provide a more comprehensive cytokinesis block micronucleus assay for biomonitoring purposes. Mutagenesis 16:51–58

    Article  PubMed  CAS  Google Scholar 

  • Kuroki M, Hachimine K, Abe H et al (2007) Sonodynamic therapy of cancer using novel sonosensitizers. Anticancer Res 27:3673–3677

    PubMed  CAS  Google Scholar 

  • Marchioni C, Riccardi E, Spinelli S, Dell’Unto F, Grimaldi P, Bedini A, Giliberti C, Giuliani L, Palomba R, Congiu Castellano A (2009) Structural changes induced in proteins by therapeutic ultrasounds. Ultrasonics 49:569–576

    Article  PubMed  CAS  Google Scholar 

  • Pozzi D, Grimaldi P, Gaudenzi S, Di Giambattista L, Silvestri I, Morrone S, Congiu Castellano A (2007) UVB-radiation-induced apoptosis in jurkat cells: a coordinated fourier transform infrared spectroscopy-flow cytometry study. Rad Res 168:698–705

    Article  CAS  Google Scholar 

  • Rosenthal I, Sostaric JZ, Riesz P (2004) Sonodynamic therapy—a review of the synergistic effects of drugs and ultrasound. Ultrasound Sonochem 11:349–363

    CAS  Google Scholar 

  • Shohet RV, Chen S, Zhou YT, Wang Z, Meidell RS, Unger RH, Grayburn PA (2000) Echocardiographic destruction of albumin microbubbles directs gene delivery to the myocardium. Circulation 101:2554–2556

    PubMed  CAS  Google Scholar 

  • Strom-Jensen PR, Dunn F (1984) Ultrasonic absorption by solvent–solute interaction and proton transfer in aqueous solutions of peptides and small proteins. J Acoust Soc Am 75:960–966

    Article  CAS  Google Scholar 

  • Wang JF, Wu CJ, Zhang CM, Qiu QY, Zheng M (2009) Ultrasound-mediated microbubble destruction facilitates gene transfection in rat C6 glioma cells. Mol Biol Rep 36:1263–1267

    Article  PubMed  CAS  Google Scholar 

  • Wang J, Guo Y, Gao J, Jin X, Wang Z, Wang B, Li K, Li Y (2010) Detection and comparison of reactive oxygen species (ROS) generated by chlorophyllin metal (Fe, Mg and Cu) complexes under ultrasonic and visible-light irradiation. Ultrasound sonochem 18(5):1028–1034. doi:10.1016/j.ultsonch.2010.12.006

    Google Scholar 

  • Xiang J, Xia X, Jiang Y, Leung AW, Wang X, Xu J, Wang P, Yu H, Bai D, Xu C (2010) Apoptosis of ovarian cancer cells induced by methylene blue-mediated sonodynamic action. Ultrasonics 51:390–395

    Article  PubMed  Google Scholar 

  • Yow CM, Wong CK, Huang Z, Ho RJ (2007) Study of the efficacy and mechanism of ALA-mediated photodynamic therapy on human hepatocellular carcinoma cell. Liver Int 27:201–208

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Experimental Medicine Department, Sapienza, University of Rome, V.le Regina Elena, 324, 00161, Rome, Italy

    D. Pozzi

  2. ISS, V.le Regina Elena, 299, 00161, Rome, Italy

    P. Fattibene & D. Viscomi

  3. Physics Department, Sapienza, University of Rome, P. le A. Moro, 2, 00185, Rome, Italy

    L. Di Giambattista, P. Grimaldi & A. Congiu Castellano

  4. CISB (Interdepartmental Research Centre for Biomedical Systems Models and Information Analysis), Rome, Italy

    L. Di Giambattista

  5. DIPIA, INAIL/ISPESL, via Urbana 167, Rome, Italy

    I. Udroiu, A. Bedini, C. Giliberti & R. Palomba

Authors
  1. D. Pozzi
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. P. Fattibene
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. D. Viscomi
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. L. Di Giambattista
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. P. Grimaldi
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. I. Udroiu
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. A. Bedini
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. C. Giliberti
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. R. Palomba
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. A. Congiu Castellano
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to D. Pozzi.

Additional information

Special Issue: SIBPA 2011 Meeting.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Pozzi, D., Fattibene, P., Viscomi, D. et al. Use of EPR and FTIR to detect biological effects of ultrasound and microbubbles on a fibroblast cell line. Eur Biophys J 40, 1115–1120 (2011). https://doi.org/10.1007/s00249-011-0738-8

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00249-011-0738-8

Keywords

Search

Navigation