Document Type : Original Article

Authors

1 Department of Radiology Technology, Faculty of Allied Medical Sciences, Shahid Beheshti University of Medical Sciences, Tehran, Iran

2 Department of Biology and Anatomical Sciences, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran

3 Department of Medical Physics and Biomedical Engineering, Tehran University of Medical Sciences, Tehran, Iran

4 Urology and Nephrology Research Center, Student Research Committee, Shahid Beheshti University of Medical Sciences, Tehran, Iran

5 Department of biostatistics and proteomics research center, Faculty of Allied Medical Sciences, Shahid Beheshti University of Medical Sciences, Tehran, Iran

Abstract

Background: Diagnostic ultrasound has been used to detect human disease especially fetus abnormalities in recent decades. Although the harmful effects of diagnostic ultrasound on human have not been established so far, several researchers showed it has had bioeffects in cell lines and in experimental animals. Three-dimensional (3D), four-dimensional (4D), and color Doppler sonography are new techniques which are widely used in diagnostic fetal ultrasonography.Objective: The study aims to evaluate some bioeffects of 3D, 4D, and color Doppler sonography in different exposure times according to the acoustic output which is set as ultrasound scanner’s default for fetal sonography in the second trimester on human dermal fibroblast (HDF) cells.Material and Methods: Exposure times selected consist of 10, 40, 70, and 100 seconds for 3D sonography, 10, 20, and 30 minutes for 4D sonography, and 10, 30, and 50 seconds for color Doppler. Cell viability, cell proliferation, and apoptosis induction on HDF cells were assessed using MTT assay, immunocytochemistry of Ki-67, and Terminal Transferase-mediated dUTP End-labeling (TUNEL) assay, respectively.Results: Exposure of cells to 3D, 4D, and color Doppler modes led to decreased cell viability and increased proliferation rate of HDF. None of the diagnostic ultrasound modes induced cell apoptosis. .Conclusion: The results indicated that 3D, 4D, and color Doppler techniques may affect the cell viability and proliferation of HDF cells, however, have no effects on the induction of apoptosis probability. Further long-term studies with other molecular endpoints are required.

Keywords

  1. Troxclair L, Smetherman D, Bluth EI. Shades of gray: a history of the development of diagnostic ultrasound in a large multispecialty clinic. The Ochsner Journal. 2011;11:151-5.
  2. Wagai T. Studies on the foundation and development of diagnostic ultrasound. Proc Jpn Acad Ser B Phys Biol Sci. 2007;83:256-65. doi: 10.2183/pjab/83.256. PubMed PMID: 24367150; PubMed Central PMCID: PMC3859294.
  3. Newnham JP, Doherty DA, Kendall GE, Zubrick SR, Landau LL, Stanley FJ. Effects of repeated prenatal ultrasound examinations on childhood outcome up to 8 years of age: follow-up of a randomised controlled trial. The Lancet. 2004;364:2038-44.doi: 10.1016/s0140-6736(04)17516-8.
  4. Udroiu I, Domenici F, Giliberti C, Bedini A, Palomba R, Luongo F, et al. Potential genotoxic effects of low-intensity ultrasound on fibroblasts, evaluated with the cytokinesis-block micronucleus assay. Mutation Research/Genetic Toxicology and Environmental Mutagenesis. 2014;772:20-4.doi: 10.1016/j.mrgentox.2014.07.004.
  5. Li J, Zhang L. Bioeffects of Diagnostic Dynamic 3-Dimensional (4-Dimensional) Ultrasound on Ultrastructure of Cerebral Cells of Fetal Mice in Late Pregnancy. Ultrasound Q. 2016;32:296-301. doi: 10.1097/RUQ.0000000000000216. PubMed PMID: 26808170.
  6. Schneider-Kolsky ME, Ayobi Z, Lombardo P, Brown D, Kedang B, Gibbs ME. Ultrasound exposure of the foetal chick brain: effects on learning and memory. Int J Dev Neurosci. 2009;27:677-83. doi: 10.1016/j.ijdevneu.2009.07.007. PubMed PMID: 19664703.
  7. Pellicer B, Herraiz S, Taboas E, Felipo V, Simon C, Pellicer A. Ultrasound bioeffects in rats: quantification of cellular damage in the fetal liver after pulsed Doppler imaging. Ultrasound Obstet Gynecol. 2011;37:643-8. doi: 10.1002/uog.8842. PubMed PMID: 20878673.
  8. Zeqiri B. Exposure criteria for medical diagnostic ultrasound: II. Criteria based on all known mechanisms:(NCRP Report No. 140) National Council on Radiation Protection and Measurements (NCRP), 2002. Ultrasound Med Biol. 2003;29:1809.
  9. Van Wamel A, Bouakaz A, ten Cate F, de Jong N, editors. Effects of diagnostic ultrasound parameters on molecular uptake and cell viability. 8-11 Oct. 2002. Munich; IEEE Ultrasonics Symposium, 2002 Proceedings.
  10. Cibull SL, Harris GR, Nell DM. Trends in diagnostic ultrasound acoustic output from data reported to the US Food and Drug Administration for device indications that include fetal applications. J Ultrasound Med. 2013;32:1921-32.doi: 10.7863/ultra.32.11.1921.
  11. Fowlkes JB, Bioeffects Committee of the American Institute of Ultrasound in M. American Institute of Ultrasound in Medicine consensus report on potential bioeffects of diagnostic ultrasound: executive summary. J Ultrasound Med. 2008;27:503-15.doi: 10.7863/jum.2008.27.4.503 . PubMed PMID: 18359906.
  12. Lai J, Pittelkow MR. Physiological effects of ultrasound mist on fibroblasts. Int J Dermatol. 2007;46:587-93. doi: 10.1111/j.1365-4632.2007.02914.x. PubMed PMID: 17550557.
  13. Franco de Oliveira R, Pires Oliveira DA, Soares CP. Effect of low-intensity pulsed ultrasound on l929 fibroblasts. Arch Med Sci. 2011;7:224-9. doi: 10.5114/aoms.2011.22071. PubMed PMID: 22291760; PubMed Central PMCID: PMC3258710.
  14. Zhou S, Schmelz A, Seufferlein T, Li Y, Zhao J, Bachem MG. Molecular mechanisms of low intensity pulsed ultrasound in human skin fibroblasts. J Biol Chem. 2004;279:54463-9. doi: 10.1074/jbc.M404786200. PubMed PMID: 15485877.
  15. Stockert JC, Horobin RW, Colombo LL, Blazquez-Castro A. Tetrazolium salts and formazan products in Cell Biology: Viability assessment, fluorescence imaging, and labeling perspectives. Acta Histochem. 2018;120:159-67. doi: 10.1016/j.acthis.2018.02.005. PubMed PMID: 29496266.
  16. Scholzen T, Gerdes J. The Ki-67 protein: from the known and the unknown. J Cell Physiol. 2000;182:311-22. doi: 10.1002/(SICI)1097-4652(200003)182:3<311::AID-JCP1>3.0.CO;2-9. PubMed PMID: 10653597.
  17. Darzynkiewicz Z, Galkowski D, Zhao H. Analysis of apoptosis by cytometry using TUNEL assay. Methods. 2008;44:250-4. doi: 10.1016/j.ymeth.2007.11.008. PubMed PMID: 18314056; PubMed Central PMCID: PMC2295206.
  18. Dinno MA, Dyson M, Young SR, Mortimer AJ, Hart J, Crum LA. The significance of membrane changes in the safe and effective use of therapeutic and diagnostic ultrasound. Phys Med Biol. 1989;34:1543-52.doi: 10.1088/0031-9155/34/11/003 . PubMed PMID: 2685832.
  19. al-Karmi AM, Dinno MA, Stoltz DA, Crum LA, Matthews JC. Calcium and the effects of ultrasound on frog skin. Ultrasound Med Biol. 1994;20:73-81. doi: 10.1016/0301-5629(94)90019-1. PubMed PMID: 8197629.
  20. Mortimer AJ, Dyson M. The effect of therapeutic ultrasound on calcium uptake in fibroblasts. Ultrasound Med Biol. 1988;14:499-506. PubMed PMID: 3227573.
  21. Ma H, Huang L, Jia J, He R, Luo L, Zhu W. Effect of energy-gathered ultrasound on Alcalase. Ultrason Sonochem. 2011;18:419-24. doi: 10.1016/j.ultsonch.2010.07.014. PubMed PMID: 20709588.
  22. De Deyne PG, Kirsch-Volders M. In vitro effects of therapeutic ultrasound on the nucleus of human fibroblasts. Phys Ther. 1995;75:629-34.
  23. Hanson MA. Health effects of exposure to ultrasound and infrasound: Report of the independent advisory group on non-ionising radiation. Health Protection Agency; 2010.
  24. Furusawa Y, Hassan MA, Zhao QL, Ogawa R, Tabuchi Y, Kondo T. Effects of therapeutic ultrasound on the nucleus and genomic DNA. Ultrason Sonochem. 2014;21:2061-8. doi: 10.1016/j.ultsonch.2014.02.028. PubMed PMID: 24657073.
  25. Young SR, Dyson M. Macrophage responsiveness to therapeutic ultrasound. Ultrasound Med Biol. 1990;16:809-16. PubMed PMID: 2095011.
  26. de Oliveira RF, Pires Oliveira DA, Machado AH, da Silva NS, Magini M, Pacheco-Soares C. Assessment of fibroblast cells submitted to ultrasonic irradiation. Cell Biol Int. 2008;32:1329-35. doi: 10.1016/j.cellbi.2008.07.016. PubMed PMID: 18703155.
  27. Izadifar Z, Babyn P, Chapman D. Mechanical and Biological Effects of Ultrasound: A Review of Present Knowledge. Ultrasound Med Biol. 2017;43:1085-104. doi: 10.1016/j.ultrasmedbio.2017.01.023. PubMed PMID: 28342566.
  28. Abramowicz JS, Barnett SB, Duck FA, Edmonds PD, Hynynen KH, Ziskin MC. Fetal thermal effects of diagnostic ultrasound. J Ultrasound Med. 2008;27:541-59. doi: 10.7863/jum.2008.27.4.541. PubMed PMID: 18359908.
  29. ter Haar G. Ultrasound bioeffects and safety. Proc Inst Mech Eng H. 2010;224:363-73. doi: 10.1243/09544119JEIM613. PubMed PMID: 20349824.