Radiation and Environmental Biophysics

, Volume 49, Issue 3, pp 397–404 | Cite as

Single exposure to radiation produces early anti-angiogenic effects in mouse aorta

  • Kevin G. Soucy
  • David O. Attarzadeh
  • Raghav Ramachandran
  • Patricia A. Soucy
  • Lewis H. Romer
  • Artin A. Shoukas
  • Dan E. BerkowitzEmail author
Original Paper


Radiation exposure can increase the risk for many non-malignant physiological complications, including cardiovascular disease. We have previously demonstrated that ionizing radiation can induce endothelial dysfunction, which contributes to increased vascular stiffness. In this study, we demonstrate that gamma radiation exposure reduced endothelial cell viability or proliferative capacity using an in vitro aortic angiogenesis assay. Segments of mouse aorta were embedded in a Matrigel-media matrix 1 day after mice received whole-body gamma irradiation between 0 and 20 Gy. Using three-dimensional phase contrast microscopy, we quantified cellular outgrowth from the aorta. Through fluorescent imaging of embedded aortas from Tie2GFP transgenic mice, we determined that the cellular outgrowth is primarily of endothelial cell origin. Significantly less endothelial cell outgrowth was observed in aortas of mice receiving radiation of 5, 10, and 20 Gy radiation, suggesting radiation-induced endothelial injury. Following 0.5 and 1 Gy doses of whole-body irradiation, reduced outgrowth was still detected. Furthermore, outgrowth was not affected by the location of the aortic segments excised along the descending aorta. In conclusion, a single exposure to gamma radiation significantly reduces endothelial cell outgrowth in a dose-dependent manner. Consequently, radiation exposure may inhibit re-endothelialization or angiogenesis after a vascular injury, which would impede vascular recovery.


Gamma Radiation Endothelial Progenitor Cell Endothelial Cell Outgrowth Angiogenic Potential Mouse Aorta 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.



We would like to thank Dr. Aleksander S. Popel, Dr. Emmanouil D. Karagiannis, and Jacob Koskimaki of Johns Hopkins University for their assistance with the aortic angiogenesis assay. This research is supported largely in part by grants from the National Aeronautics and Space Administration (NNJ05HF03G) and National Space Biomedical Research Institute (NCC 9-58-CA01301).


  1. Aplin AC, Fogel E, Zorzi P, Nicosia RF (2008) The aortic ring model of angiogenesis. Methods Enzymol 443:119–136CrossRefGoogle Scholar
  2. Asahara T, Murohara T, Sullivan A, Silver M, van der Zee R, Li T, Witzenbichler B, Schatteman G, Isner JM (1997) Isolation of putative progenitor endothelial cells for angiogenesis. Science 275:964–967CrossRefGoogle Scholar
  3. Benderitter M, Maingon P, Abadie C, Assem M, Maupoil V, Briot F, Horiot JC, Rochette L (1995) Effect of in vivo heart irradiation on the development of antioxidant defenses and cardiac functions in the rat. Radiat Res 144:64–72CrossRefGoogle Scholar
  4. Brooks A, Bao S, Rithidech K, Couch LA, Braby LA (2001) Relative effectiveness of hze iron-56 particles for the induction of cytogenetic damage in vivo. Radiat Res 155:353–359CrossRefGoogle Scholar
  5. Cardis E, Gilbert ES, Carpenter L, Howe G, Kato I, Armstrong BK, Beral V, Cowper G, Douglas A, Fix J et al (1995) Effects of low doses and low dose rates of external ionizing radiation: cancer mortality among nuclear industry workers in three countries. Radiat Res 142:117–132CrossRefGoogle Scholar
  6. Carmeliet P (2005) Angiogenesis in life, disease and medicine. Nature 438:932–936CrossRefADSGoogle Scholar
  7. Cucinotta FA, Durante M (2006) Cancer risk from exposure to galactic cosmic rays: implications for space exploration by human beings. Lancet Oncol 7:431–435CrossRefGoogle Scholar
  8. De Bruin ML, Dorresteijn LD, van’t Veer MB, Krol AD, van der Pal HJ, Kappelle AC, Boogerd W, Aleman BM, van Leeuwen FE (2009) Increased risk of stroke and transient ischemic attack in 5-year survivors of hodgkin lymphoma. J Natl Cancer Inst 101:928–937CrossRefGoogle Scholar
  9. Denham JW, Hauer-Jensen M (2002) The radiotherapeutic injury: a complex “wound”. Radiother Oncol 63:129–145CrossRefGoogle Scholar
  10. Denham JW, Peters LJ, Johansen J, Poulsen M, Lamb DS, Hindley A, O’Brien PC, Spry NA, Penniment M, Krawitz H, Williamson S, Bear J, Tripcony L (1999) Do acute mucosal reactions lead to consequential late reactions in patients with head and neck cancer? Radiother Oncol 52:157–164CrossRefGoogle Scholar
  11. Dimmeler S, Zeiher AM (1997) Nitric oxide and apoptosis: another paradigm for the double-edged role of nitric oxide. Nitric Oxide 1:275–281CrossRefGoogle Scholar
  12. Dorresteijn LD, Kappelle AC, Boogerd W, Klokman WJ, Balm AJ, Keus RB, van Leeuwen FE, Bartelink H (2002) Increased risk of ischemic stroke after radiotherapy on the neck in patients younger than 60 years. J Clin Oncol 20:282–288CrossRefGoogle Scholar
  13. EBCTCG (2000) Favourable and unfavourable effects on long-term survival of radiotherapy for early breast cancer: an overview of the randomised trials. Early breast cancer trialists’ collaborative group. Lancet 355:1757–1770CrossRefGoogle Scholar
  14. Gelati M, Aplin AC, Fogel E, Smith KD, Nicosia RF (2008) The angiogenic response of the aorta to injury and inflammatory cytokines requires macrophages. J Immunol 181:5711–5719Google Scholar
  15. Gratwohl A, John L, Baldomero H, Roth J, Tichelli A, Nissen C, Lyman SD, Wodnar-Filipowicz A (1998) Flt-3 ligand provides hematopoietic protection from total body irradiation in rabbits. Blood 92:765–769Google Scholar
  16. Hancock SL, Tucker MA, Hoppe RT (1993) Factors affecting late mortality from heart disease after treatment of hodgkin’s disease. JAMA 270:1949–1955CrossRefGoogle Scholar
  17. Hillen F, Kaijzel EL, Castermans K, Oude Egbrink MG, Lowik CW, Griffioen AW (2008) A transgenic tie2-gfp athymic mouse model; a tool for vascular biology in xenograft tumors. Biochem Biophys Res Commun 368:364–367CrossRefGoogle Scholar
  18. Kollum M, Cottin Y, Chan RC, Kim HS, Bhargava B, Vodovotz Y, Waksman R (2001) Decreased adventitial neovascularization after intracoronary irradiation in swine: a time course study. Int J Radiat Oncol Biol Phys 50:1033–1039CrossRefGoogle Scholar
  19. Lauk S (1987) Endothelial alkaline phosphatase activity loss as an early stage in the development of radiation-induced heart disease in rats. Radiat Res 110:118–128CrossRefGoogle Scholar
  20. Lauk S, Kiszel Z, Buschmann J, Trott KR (1985) Radiation-induced heart disease in rats. Int J Radiat Oncol Biol Phys 11:801–808Google Scholar
  21. Li J, Zhang YP, Kirsner RS (2003) Angiogenesis in wound repair: angiogenic growth factors and the extracellular matrix. Microsc Res Tech 60:107–114CrossRefGoogle Scholar
  22. Montesinos MC, Shaw JP, Yee H, Shamamian P, Cronstein BN (2004) Adenosine a(2a) receptor activation promotes wound neovascularization by stimulating angiogenesis and vasculogenesis. Am J Pathol 164:1887–1892Google Scholar
  23. Oh CW, Bump EA, Kim JS, Janigro D, Mayberg MR (2001) Induction of a senescence-like phenotype in bovine aortic endothelial cells by ionizing radiation. Radiat Res 156:232–240CrossRefGoogle Scholar
  24. On YK, Kim HS, Kim SY, Chae IH, Oh BH, Lee MM, Park YB, Choi YS, Chung MH (2001) Vitamin c prevents radiation-induced endothelium-dependent vasomotor dysfunction and de-endothelialization by inhibiting oxidative damage in the rat. Clin Exp Pharmacol Physiol 28:816–821CrossRefGoogle Scholar
  25. Preston DL, Shimizu Y, Pierce DA, Suyama A, Mabuchi K (2003) Studies of mortality of atomic bomb survivors. Report 13: solid cancer and noncancer disease mortality: 1950-1997. Radiat Res 160:381–407CrossRefGoogle Scholar
  26. Qi F, Sugihara T, Hattori Y, Yamamoto Y, Kanno M, Abe K (1998) Functional and morphological damage of endothelium in rabbit ear artery following irradiation with cobalt60. Br J Pharmacol 123:653–660CrossRefGoogle Scholar
  27. Rose RW, Grant DS, O’Hara MD, Williamson SK (1999) The role of laminin-1 in the modulation of radiation damage in endothelial cells and differentiation. Radiat Res 152:14–28CrossRefGoogle Scholar
  28. Sieveking DP, Buckle A, Celermajer DS, Ng MK (2008) Strikingly different angiogenic properties of endothelial progenitor cell subpopulations: insights from a novel human angiogenesis assay. J Am Coll Cardiol 51:660–668CrossRefGoogle Scholar
  29. Sonveaux P, Brouet A, Havaux X, Gregoire V, Dessy C, Balligand JL, Feron O (2003) Irradiation-induced angiogenesis through the up-regulation of the nitric oxide pathway: implications for tumor radiotherapy. Cancer Res 63:1012–1019Google Scholar
  30. Soucy KG, Lim HK, Benjo A, Santhanam L, Ryoo S, Shoukas AA, Vazquez ME, Berkowitz DE (2007) Single exposure gamma-irradiation amplifies xanthine oxidase activity and induces endothelial dysfunction in rat aorta. Radiat Environ Biophys 46:179–186Google Scholar
  31. Spees JL, Whitney MJ, Sullivan DE, Lasky JA, Laboy M, Ylostalo J, Prockop DJ (2008) Bone marrow progenitor cells contribute to repair and remodeling of the lung and heart in a rat model of progressive pulmonary hypertension. Faseb J 22:1226–1236CrossRefGoogle Scholar
  32. Stewart JR, Fajardo LF (1971) Radiation-induced heart disease. Clinical and experimental aspects. Radiol Clin North Am 9:511–531Google Scholar
  33. Sugihara T, Hattori Y, Yamamoto Y, Qi F, Ichikawa R, Sato A, Liu MY, Abe K, Kanno M (1999) Preferential impairment of nitric oxide-mediated endothelium-dependent relaxation in human cervical arteries after irradiation. Circulation 100:635–641Google Scholar
  34. Urbich C, Dimmeler S (2004) Endothelial progenitor cells: characterization and role in vascular biology. Circ Res 95:343–353CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  • Kevin G. Soucy
    • 1
  • David O. Attarzadeh
    • 1
  • Raghav Ramachandran
    • 1
  • Patricia A. Soucy
    • 1
  • Lewis H. Romer
    • 2
  • Artin A. Shoukas
    • 1
  • Dan E. Berkowitz
    • 2
    Email author
  1. 1.Biomedical EngineeringJohns Hopkins UniversityBaltimoreUSA
  2. 2.Anesthesiology and Critical Care MedicineJohns Hopkins Medical InstitutionsBaltimoreUSA

Personalised recommendations